ovalbumin and Asthma

Allergic asthma is an inflammatory disease caused by the immune system's reaction to allergens, inflammation and narrowing of the airways, and the production of more than normal mucus. One of the main reasons is an increased production of inflammatory cytokines in the lungs that leads to the appearance of symptoms of asthma, including inflammation and shortness of breath. On the other hand, it has been proven that phytochemicals with their antioxidant and anti-inflammatory properties can be useful in improving allergic asthma.. Common chemical treatments for allergic asthma include corticosteroids, which have many side effects and temporarily relieve symptoms but are not a cure. Therefore, taking the help of natural compounds to improve the quality of life of asthmatic patients can be a valuable issue that has been evaluated in the present review.. In this study, three databases (Scopus, PubMed, and Cochrane) with the keywords: allergic asthma, phytochemical, plant, and herb were evaluated. The primary result was 5307 articles. Non-English, repetitive, and review articles were deleted from the study.. Finally, after carefully reading the articles, 102 were included in the study (2006-2022). The results of this review state that phytochemicals suppress the inflammatory pathways via inhibition of inflammatory cytokines production/secretion, genes, and proteins involved in the inflammation process, reducing oxidative stress indicators and symptoms of allergic asthma, such as cough and mucus production in the lungs.. With their antioxidant effects, this study concluded that phytochemicals suppress cytokines and other inflammatory indicators and thus can be considered an adjunctive treatment for improving allergic asthma.

Topics: Animals; Antioxidants; Asthma; Cytokines; Disease Models, Animal; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phytochemicals; Quality of Life

Allergic diseases, in particular atopic asthma, have been on the rise in most industrialized countries for several decades now. Allergic asthma is characterized by airway narrowing, bronchial hyperresponsiveness, excessive airway mucus production, eosinophil influx in the lungs and an imbalance of the Th1/Th2 responses, including elevated IgE levels. Most available interventions provide only short-term relief from disease symptoms and do not alter the underlying immune imbalance. A number of studies, mostly in mouse models, have shown that Mycobacterium bovis bacillus Calmette-Guérin (BCG) treatment is capable of preventing or reducing an established allergen-driven inflammatory response, by redirecting pathogenic Th2 towards protective Th1 and/or regulatory T cell responses. Dendritic cells stimulated by BCG appear to be a crucial first step in the immunomodulatory effects of BCG. While the protective and therapeutic effects of BCG against allergy and asthma are well documented in animal models, they are less clear in humans, both in observational studies and in randomized controlled trials. The purpose of this article is to provide an up-to-date overview of the available evidence on the anti-allergy, in particular anti-asthma effects of BCG in mice, rats and humans.

Topics: Animals; Asthma; BCG Vaccine; Hypersensitivity; Lung; Mice; Mycobacterium bovis; Ovalbumin; Rats; Th2 Cells

Asthma is an important issue not only in health but also in economics worldwide. Therefore, asthma animal models have been frequently used to understand the pathogenesis of asthma. Recently, in addition to acquired immunity, innate immunity has also been thought to be involved in asthma. Among innate immune cells, group 2 innate lymphoid cells (ILC2s) have been considered to be crucial for eosinophilic airway inflammation by releasing T helper 2 cytokines. Moreover, house dust mites (HDMs) belonging to group 1 act on airway epithelial cells not only as allergens but also as cysteine proteases. The production of interleukin-25 (IL-25), IL-33, and thymic stromal lymphopoietin (TSLP) from airway epithelial cells was induced by the protease activity of HDMs. These cytokines activate ILC2s, and activated ILC2s produce IL-5, IL-9, IL-13, and amphiregulin. Hence, the HDM-induced asthma mouse model greatly contributes to understanding asthma pathogenesis. In this review, we highlight the relationship between ILC2s and the HDM in the asthma mouse model to help researchers and clinicians not only choose a proper asthma mouse model but also to understand the molecular mechanisms underlying HDM-induced asthma.

Topics: Animals; Aspergillus fumigatus; Asthma; Disease Models, Animal; Humans; Immunity, Innate; Lymphocytes; Mice; Ovalbumin; Pyroglyphidae

Asthma is a heterogeneous disease with increasing prevalence worldwide characterized by chronic airway inflammation, increased mucus secretion and bronchial hyperresponsiveness. The phenotypic heterogeneity among asthmatic patients is accompanied by different endotypes, mainly Type 2 or non-Type 2. To investigate the pathomechanism of this complex disease many animal models have been developed, each trying to mimic specific aspects of the human disease. Rodents have classically been employed in animal models of asthma. The present review provides an overview of currently used Type 2 vs. non-Type 2 rodent asthma models, both acute and chronic. It further assesses the methods used to simulate disease development and exacerbations as well as to quantify allergic airway inflammation, including lung physiologic, cellular and molecular immunologic responses. Furthermore, the employment of genetically modified animals, which provide an in-depth understanding of the role of a variety of molecules, signaling pathways and receptors implicated in the development of this disease as well as humanized models of allergic inflammation, which have been recently developed to overcome differences between the rodent and human immune systems, are discussed. Nevertheless, differences between mice and humans should be carefully considered and limits of extrapolation should be wisely taken into account when translating experimental results into clinical use.

Topics: Acute Disease; Airway Remodeling; Aluminum Hydroxide; Animals; Antigens; Asthma; Bronchoconstriction; Chronic Disease; Disease Models, Animal; Humans; Inflammation Mediators; Lung; Ovalbumin; Signal Transduction; Species Specificity

Asthma is a chronic inflammatory disease of the airway with extensive airway remodeling. The ethical issues associated with the studies in asthmatic patients, required development of animal model of asthma. Animal models of asthma can provide valuable information on several features of asthma pathogenesis and treatment. Although these models cannot carry out all clinical features, they are valuable to understand mechanisms of the disease and curative access. Related articles were searched in different databases from September 1994 to April 2016 using; animal model of asthma, animal sensitization, allergen-induced asthma in animals terms. Although there are several reviews on this topic, in the present article, induction of animal model of asthma in different animals, various methods used for this purpose, measured parameters and research purposes were reviewed, which will help investigators to use the appropriate animal, methods, and evaluating parameters depending on their study design. In this study various method used for induction of animal model of asthma in different animals and measured parameters were described, which will help investigators to use the appropriate animal, method and evaluating parameters depending on their study design.

Topics: Airway Remodeling; Allergens; Animals; Asthma; Disease Models, Animal; Guinea Pigs; Immunization; Inflammation; Mice; Ovalbumin; Rabbits; Rats; Respiratory Hypersensitivity

Asthma is a heterogeneous disease in which various environmental stimuli as well as different genes, cell types, cytokines and mediators are implicated. This chronic inflammatory disorder of the airways is estimated to affect as many as 300 million people worldwide. Animal models of asthma, despite their limitations, have contributed greatly to our understanding of disease pathology and the identification of key processes, cells and mediators in asthma. However, it is less likely to develop an animal model of asthma that takes into account all aspects of human disease. The focus in current asthma research is increasingly on severe asthma because this group of patients is not well treated today. Recent advances in studies of asthma exacerbation are thus considered. We therefore need to develop translational model systems for pharmacological evaluation and molecular target discovery of severe asthma and asthma exacerbations. In this review we attempted to discuss the different animal models of asthma, with special emphasis on ovalbumin and house dust mite models, their merits and their limitations.

Topics: Animals; Antigens, Dermatophagoides; Asthma; Disease Models, Animal; Disease Progression; Humans; Ovalbumin; Species Specificity; Translational Research, Biomedical

There is little data on the effect of exercise on markers of airway inflammation in human asthmatics. The main objective of this review is to determine the effects of physical training on markers of airway inflammation in animal models of asthma.. A peer reviewed search was applied to Medline, Embase, Web of Science, Cochrane, and DARE databases. Data extraction was performed in a blinded fashion.. From the initial 2336 studies, a total of 10 studies were selected for the final analysis. All were randomized controlled trials with low to moderate intensity training on ovalbumin-sensitized mice. In the exercised group of mice, there was a reduction in BAL eosinophils and Th-2 cytokines, no change in Th-1 cytokines, an increase in IL-10, and a reversal of airway remodeling. The data was not pooled owing to significant heterogeneity between studies, and a funnel plot test for publication bias was not performed because there were few studies reporting on any one outcome measure. The asthma models differed between studies in age and gender of mice, as well as in timing of physical training after sensitization. The risk of bias was unclear for some studies though this may not influence outcome measures. The accuracy of data extracted from graphics is unknown.. Physical training improves airway inflammation in animal asthma models.

Topics: Animals; Asthma; Biomarkers; Cytokines; Disease Models, Animal; Female; Guinea Pigs; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Physical Conditioning, Animal; Pneumonia

Galectins constitute an evolutionary conserved family that bind to β-galactosides. Increasing evidence shows that galectins are involved in many fundamental biological processes such as cellular communication, inflammation, differentiation and apoptosis. Changes in galectin-3 (Gal-3) expression are commonly seen in cancer and pre-cancerous conditions, and Gal-3 may be involved in the regulation of diverse cancer cell activities that contribute to tumourigenesis, cancer progression and metastasis. In addition, Gal-3 is a pro-inflammatory regulator in rheumatoid arthritis. Gal-3 has been shown to be involved in many aspects in allergic inflammation, such as eosinophil recruitment, airway remodeling, development of a Th2 phenotype as well as increased expression of inflammatory mediators. In an in vivo model it was shown that bronchoalveolar lavage (BAL) fluid from ovalbumin-challenged mice contained significantly higher levels of Gal-3 compared to control mice. The molecular mechanisms of Gal-3 in human asthma have not been fully elucidated. This review will focus on what is known about the Gal-3 and its role in the pathophysiological mechanisms of asthma to evaluate the potential of Gal-3 as a biomarker and therapeutic target of asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Galectin 3; Mice; Ovalbumin

The present work was aimed to review modern approaches to simulation of bronchial asthma using laboratory animals. Various modes of immunization and challenge for the induction of typical pathological features in mice are described along with the methods for detection of these markers (including ones for studying murine lung function). Advantages and drawbacks of the models, difficulties and prospects of their application for the study of pathogenesis and preliminary estimation of the safety and efficacy of antiasthmatic therapy (especially antigen-specific immunotherapy) are analyzed. Moreover, the paper presents concise characteristic of short-term adjuvant-free murine models of timothy pollen extract- and ovalbumin-induced bronchial asthma developed in this country.

Topics: Allergens; Animals; Asthma; Disease Models, Animal; Immunoglobulin E; Mice; Ovalbumin; Pollen; Species Specificity

Cell-based therapies with embryonic or adult stem cells, including induced pluripotent stem cells, have emerged as potential novel approaches for several devastating and otherwise incurable lung diseases, including emphysema, pulmonary fibrosis, pulmonary hypertension, and the acute respiratory distress syndrome. Although initial studies suggested engraftment of exogenously administered stem cells in lung, this is now generally felt to be a rare occurrence of uncertain physiologic significance. However, more recent studies have demonstrated paracrine effects of administered cells, including stimulation of angiogenesis and modulation of local inflammatory and immune responses in mouse lung disease models. Based on these studies and on safety and initial efficacy data from trials of adult stem cells in other diseases, groundbreaking clinical trials of cell-based therapy have been initiated for pulmonary hypertension and for chronic obstructive pulmonary disease. In parallel, the identity and role of endogenous lung progenitor cells in development and in repair from injury and potential contribution as lung cancer stem cells continue to be elucidated. Most recently, novel bioengineering approaches have been applied to develop functional lung tissue ex vivo. Advances in each of these areas will be described in this review with particular reference to animal models.

Topics: Animals; Asthma; Bioengineering; Cell- and Tissue-Based Therapy; Disease Models, Animal; Embryonic Stem Cells; Emphysema; Humans; Hypertension, Pulmonary; Lung; Lung Diseases; Mice; Ovalbumin; Pulmonary Fibrosis; Regeneration; Respiratory Distress Syndrome; Stem Cell Transplantation; Stem Cells

Ovalbumin challenge models of asthma offer many opportunities for increasing our understanding of the pathogenetic mechanisms underlying this disease, as well as for identifying novel therapeutic targets. There is no single "classical" model, because numerous alternatives exist with respect to the choice of mouse strain, method of sensitisation, route and duration of challenge, and approach to assessing the host response. Moreover, the limitations of these models need to be recognised when attempting to interpret experimental findings. Nevertheless, careful use of well-defined models allows investigators to answer specific questions that are otherwise difficult to address.

Topics: Animals; Asthma; Disease Models, Animal; Drug Evaluation, Preclinical; Humans; Immunization; Mice; Mice, Inbred Strains; Ovalbumin; Respiratory Hypersensitivity

Allergic asthma is defined as a hypersensitivity reaction of the lung towards per se harmless antigens, e.g. pollen and house dust mite, accompanied with a chronic eosinophilic inflammation of the lung. During the course of the disease, physiological and structural changes in the lung occur, i.e. airway hyperresponsiveness, restricted airflow and airway remodelling. In addition to ovalbumin-induced mouse models of acute asthma, recently new models were developed, which show a closer resemblance to human asthma, both regarding the induction of characteristics of chronic allergic inflammation and the use of clinical relevant allergens. Moreover, attention is paid on the influence of adjuvants or the route of sensitisation on the protocol outcomes. The effort spent in development of these new models will be worthwhile, especially for research in the field of immuno-therapy. These improved animal models may broaden the knowledge of the disease and thereby provide new strategies for preventive and therapeutic interventions.

Topics: Allergens; Animals; Antigens, Plant; Asthma; Disease Models, Animal; Humans; Hypersensitivity; Immunization; Mice; Ovalbumin; Plant Extracts

House dust mite (HDM) is the most pervasive indoor aeroallergen source worldwide. Allergens derived from HDM are associated with sensitization and allergic asthma. Allergic asthma is an immunologically driven disease characterized by a Th2-polarized immune response, eosinophilic inflammation, airway hyperreactivity, and remodeling. Animal models of asthma utilizing ovalbumin (OVA) exposure have afforded us considerable insight with respect to the mediators and cell types involved in allergic airway inflammation. However, OVA preparations and HDM are two vastly different materials. This chapter is specifically concerned with modeling responses to HDM exposure in mice. These studies have furnished new information and unlocked new lines of inquiry regarding biological responses to common aeroallergens. The complexity of HDM as an allergen source, with its plethora of protein and nonprotein immunogenic components, may influence the mechanisms underlying sensitization, inflammation and remodeling. Here, we will discuss this issue, along with giving critical thought to the use of experimental models.

Topics: Animals; Antigens, Dermatophagoides; Asthma; Bronchial Hyperreactivity; Disease Models, Animal; Humans; Hypersensitivity; Mice; Ovalbumin; Pyroglyphidae; Th2 Cells

Bronchial asthma is a complicated and diverse disorder affected by genetic and environmental factors. It is widely accepted that it is a Th2-type inflammation originating in lung and caused by inhalation of ubiquitous allergens. The complicated and diverse pathogenesis of this disease yet to be clarified. Functional genomics is the analysis of whole gene expression profiling under given condition, and microarray technology is now the most powerful tool for functional genomics. Several attempts to clarify the pathogenesis of bronchial asthma have been carried out using microarray technology, providing us some novel biomarkers for diagnosis, therapeutic targets or understanding pathogenic mechanisms of bronchial asthma. In this article, we review the outcomes of these analyses by the microarray approach as applied to this disease by focusing on the identification of novel biomarkers.

Topics: Animals; Antigens, Fungal; Ascaris; Aspergillus; Asthma; Biomarkers; Cytokines; Fibroblasts; Gene Expression Profiling; Gene Expression Regulation; Genetic Predisposition to Disease; Genomics; Humans; Interleukin-13; Interleukin-4; Lung; Mast Cells; Mice; Mice, Inbred A; Mice, Inbred C3H; Mice, Knockout; Oligonucleotide Array Sequence Analysis; Ovalbumin; Reverse Transcriptase Polymerase Chain Reaction; Th2 Cells

Allergic bronchial asthma is characterized by chronic inflammation of the airways, development of airway hyperreactivity and recurrent reversible airway obstruction. Target and effector cells responsible for airway hyperresponsiveness and airway obstruction include sensory and motor neurons as well as epithelial and smooth muscle cells. Although it is well established that the inflammatory process is controlled by T-helper-2 (Th2) cells, the mechanisms by which immune cells interact with neurons, epithelial cells or smooth muscle cells still remain uncertain. Due to growing evidence for extensive communication between neurons and immune cells, the mechanisms of this neuroimmune crosstalk in lung and airways of asthmatic patients are becoming the focus of asthma research. Neurotrophins represent molecules potentially responsible for regulating and controlling the crosstalk between the immune and peripheral nervous system. They are constitutively expressed by resident lung cells and produced in increasing concentrations by immune cells invading the airways under pathological conditions. Neurotrophins modify the functional activity of sensory and motor neurons, leading to enhanced and altered neuropeptide and tachykinin production. These effects are defined as neuronal plasticity. The consequences are the development of neurogenic inflammation.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Humans; Mice; Mice, Knockout; Nerve Growth Factors; Neurogenic Inflammation; Neuronal Plasticity; Ovalbumin; Rats; Receptor, Nerve Growth Factor; Receptors, Nerve Growth Factor; T-Lymphocytes, Helper-Inducer; Tachykinins; Time Factors

Nerve growth factor (NGF), in addition to its essential role in neuronal growth and survival, may also act as an inflammatory mediator. As several animal studies have shown, NGF appears to play a part in the development of airway hyperresponsiveness and in the increased sympathetic and sensory innervation of the lung. It also has a profound effect on airway inflammation and asthma-related symptoms. Sources of NGF in the airways are numerous: inflammatory cells infiltrated into the bronchial mucosa, and structural cells including lung fibroblasts, airway epithelial and smooth muscle cells. These cells, by releasing more NGF in inflammatory conditions, may contribute to the increased NGF levels observed in bronchoalveolar lavage fluid and serum from patients with asthma. Taken together, these results suggest that NGF is an important mediator in both inflammation and asthma.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Cells, Cultured; Disease Models, Animal; Dose-Response Relationship, Drug; Fibroblasts; Humans; Inflammation; Interleukin-1; Lung; Muscle, Smooth; Nerve Growth Factor; Neuroimmunomodulation; Ovalbumin; Trachea

Epidemiological associations between ozone exposure and allergic responsiveness are well-documented and have been corroborated in animal studies. The complex interaction between ozone and allergen has genetic and environmental components that affect atopic individuals and may increase the incidence of allergy in susceptible individuals. This review describes the advances that have been made in understanding mechanisms of genetic susceptibility to ozone-induced inflammation, and the interaction between ozone and allergen exposure in mice and a non-human primate model.. Antioxidant and innate immune defense genes contribute to ozone-induced inflammation and hyperpermeability in mice and humans. Ozone exposure during the allergic challenge phase induces greater enhancement of allergic responsiveness than the sensitization stage. Ovalbumin-pulsed dendritic cells injected into naïve mice successfully sensitize the mouse to ovalbumin in the absence of adjuvant. Debate continues over the role of T helper 1-T helper 2 immune profile development in mediating the ozone-allergen interaction, and the potential confounding influence of the predominant T helper 2 system most commonly used to study these responses.. The role of genetic background in susceptibility to ozone-induced lung inflammation has been confirmed, and promising candidate genes have been identified. Descriptive studies confirm that ozone exacerbates allergic responsiveness. Ozone administered during the challenge phase of ovalbumin allergen exposure induces greater responsiveness than during the sensitization phase. Allergen-induced responses enhanced by concurrent ozone exposure warrant further mechanistic research, particularly regarding the influence of susceptibility genes.

Topics: Allergens; Animals; Asthma; Genetic Predisposition to Disease; Humans; Macaca mulatta; Mice; Models, Animal; Ovalbumin; Ozone; Pneumonia

Topics: Animals; Asthma; Bronchial Provocation Tests; Collagen; Disease Models, Animal; Immunization; Mice; Ovalbumin; Respiratory Mucosa; Respiratory System; Trachea

Topics: Airway Obstruction; Animals; Asthma; Disease Models, Animal; Disease Progression; Eosinophils; Humans; Immunization; Nasal Provocation Tests; Ovalbumin

Antigen-presenting dendritic cells are essential for the recognition and presentation of allergens to the cells of the immune system. Airway dendritic cells capture allergen in the mucosa and present it to naive T cells after migration into the draining lymph nodes. In this review article, we discuss the most recent findings from animal models of asthma, which highlight an essential role for these cells in the induction and maintenance of eosinophilic airway inflammation. This increasing knowledge might lead to the identification of new targets for the prevention and therapy of asthma.

Topics: Administration, Inhalation; Allergens; Animals; Antigen Presentation; Asthma; Dendritic Cells; Disease Models, Animal; Humans; Lung; Mice; Models, Immunological; Ovalbumin; Pulmonary Eosinophilia; Rats; T-Lymphocytes

The fact that only some individuals exposed to environmental chemicals develop chemical intolerance raises the possibility that genetic factors could be contributing factors. The present communication summarizes evidence from a genetic animal model of cholinergic supersensitivity that suggests that an abnormal cholinergic system could be one predisposing genetic factor. The Flinders Sensitive Line (FSL) rats were established by selective breeding for increased responses to an organophosphate. It was subsequently found that these FSL rats were also more sensitive to direct-acting muscarinic agonists and had elevated muscarinic receptors compared to the selectively bred parallel group, the Flinders Resistant Line (FRL) rats, or randomly bred control rats. Increased sensitivity to cholinergic agents has also been observed in several human populations, including individuals suffering from chemical intolerance. Indeed, the FSL rats exhibit certain behavioral characteristics such as abnormal sleep, activity, and appetite that are similar to those reported in these human populations. In addition, the FSL rats have been reported to exhibit increased sensitivity to a variety of other chemical agents. Peripheral tissues, such as intestinal and airway smooth muscle, appear to be more sensitive to both cholinergic agonists and an antigen, ovalbumin. Hypothermia, a centrally mediated response, is more pronounced in the FSL rats after nicotine and alcohol, as well as agents that are selective for the dopaminergic and serotonergic systems. In some cases, the increased sensitivity has been detected in the absence of any changes in the receptors with which the drugs interact (dopamine receptors), while receptor changes have been seen in other cases (nicotine receptors). Therefore, there may be multiple mechanisms underlying the multiple chemical sensitivity-chemical intolerance of the FSL rats. An elucidation of these mechanisms may provide useful clues to those involved in chemical intolerance in humans.

Topics: Acetylcholine; Allergens; Animals; Asthma; Cholinergic Agents; Cholinergic Fibers; Cholinesterase Inhibitors; Disease Models, Animal; Environmental Pollutants; Fatigue Syndrome, Chronic; Humans; Hypothermia; Models, Biological; Multiple Chemical Sensitivity; Muscarinic Agonists; Muscle, Smooth; Nicotinic Agonists; Ovalbumin; Pesticides; Rats; Rats, Inbred Strains; Rats, Sprague-Dawley; Receptors, Dopamine; Receptors, Muscarinic; Receptors, Nicotinic; Second Messenger Systems; Serotonin Receptor Agonists; Sleep Wake Disorders; Up-Regulation

Topics: Allergens; Animals; Asthma; Biopsy; Blood Proteins; Bronchi; Cell Adhesion; Crosses, Genetic; Dendritic Cells; Eosinophil Granule Proteins; Eosinophils; Epithelial Cells; Fibroblasts; Humans; Inflammation Mediators; Matrix Metalloproteinase 9; Mice; Mice, Transgenic; Muscle, Smooth; Ovalbumin; Pulmonary Eosinophilia; Respiratory Tract Infections; Ribonucleases; Th1 Cells; Th2 Cells; Tissue Inhibitor of Metalloproteinase-1; Transgenes; Virus Diseases

Topics: Antigens; Asthma; Epitopes; Humans; Immunoglobulin E; Immunoglobulin G; Immunotherapy; Ovalbumin; T-Lymphocytes

Topics: Animals; Asthma; Guinea Pigs; Humans; Lung; Muscle, Smooth; Nucleosides; Nucleotides; Ovalbumin; Respiratory System; Uridine Triphosphate

Cytokines are very potent pro-inflammatory agents. Several cytokines are present in abnormal quantities in asthmatic airway tissues. In vitro and in vivo experiments demonstrate that these cytokines have biological effects relevant to the pathogenesis of asthma. We review the evidence that interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-10 (IL-10), interleukin-12 (IL-12) and interferon-gamma (IFNgamma) have the potential of playing a key role in the pathogenesis of asthma. Inhibition of the activity of IL-4 or IL-5 and enhancing or mimicking the action of IL-10, IL-12 or IFNgamma are therapeutic options in asthma that warrant further investigations.

Topics: Administration, Inhalation; Animals; Antibody Specificity; Asthma; Cytokines; Disease Models, Animal; Hypersensitivity, Immediate; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-4; Interleukin-5; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Receptors, Cytokine; Respiratory System

Haemophilus influenzae is a bacterium that often can be isolated from the deeper respiratory airways of patients with chronic asthmatic bronchitis. In the present study the effects of H. influenzae vaccination on guinea pig pulmonary beta-adrenoceptor number and function (in vitro and in vivo) have been evaluated. Functioning of beta-adrenoceptors is determined by measuring the beta-mimetic effect of isoprenaline on the inhibition of anaphylactic mediator release and isolated tracheal strip relaxation. The number of beta-adrenoceptor binding sites was measured by means of a 3H-dihydroalprenolol binding assay. Also the mechanism of action underlying the changes in beta-adrenoceptor functioning was evaluated. Furthermore, it was established that the effect on the beta-adrenoceptor system was not specific for H. influenzae and that other respiratory pathogens were also biologically active in this respect.

Topics: Animals; Antigens, Bacterial; Asthma; Carbachol; Guinea Pigs; Haemophilus influenzae; Histamine Release; Humans; Isoproterenol; Lung; Lung Diseases, Obstructive; Ovalbumin; Polysaccharides, Bacterial; Receptors, Adrenergic; Receptors, Adrenergic, beta; Trachea

Topics: Adolescent; Adult; Allergens; Animals; Antibody Formation; Asthma; Binding Sites, Antibody; Cattle; Child; Chromosome Mapping; Disease Models, Animal; Diseases in Twins; Eczema; Female; gamma-Globulins; Genes, MHC Class II; Genotype; Haploidy; Histocompatibility Antigens; Humans; Hypersensitivity, Immediate; Immunity; Immunoglobulin E; Immunoglobulin G; In Vitro Techniques; Male; Mice; Mice, Inbred Strains; Middle Aged; Ovalbumin; Passive Cutaneous Anaphylaxis; Pedigree; Penicillin G; Pollen; Reagins; Rhinitis, Allergic, Seasonal

The proinflammatory cytokine interleukin-17A (IL-17A) is known to mediate antimicrobial activity, but its role during rhinovirus (RV) infections and in asthma needs further investigation. Therefore, we addressed the role of IL-17A during allergic asthma and antiviral immune response in human and murine immunocompetent cells. In this study we found that asthmatic children with a RV infection in their upper airways have upregulated mRNA levels of the antiviral cytokine interferon type I (IFN)-β and the transcription factor T-box 21 (TBX21) and reduced levels of IL-17A protein in their peripheral blood mononuclear cells (PBMCs). We also found that IL-17A inhibited RV1b replication in infected human lung epithelial cells A549. Furthermore, by using gene array analysis we discovered that targeted deletion of Il17a in murine lung CD4(+) T cells impaired Oas1g mRNA downstream of Ifnβ, independently from RV infection. Additionally, in PBMCs of children with a RV infection in their nasalpharyngeal fluid OAS1 gene expression was found downregulated. Finally RV1b inhibited IL-17A production in lung CD4(+) T cells in a setting of experimental asthma. These results indicate that the RV1b inhibits IL-17A in T helper type 17 cells and IL-17A clears RV1b infection in epithelial cells. In both cases IL-17A contributes to fend off RV1b infection by inducing genes downstream of interferon type I pathway.

Topics: 2',5'-Oligoadenylate Synthetase; A549 Cells; Animals; Asthma; CD4-Positive T-Lymphocytes; Child; Child, Preschool; Drug Hypersensitivity; Female; Gene Expression Regulation; Humans; Interferon-beta; Interleukin-17; Lung; Male; Mice; Mice, Knockout; Ovalbumin; Picornaviridae Infections; Primary Cell Culture; Rhinovirus; RNA, Messenger; Signal Transduction; T-Box Domain Proteins

The histamine H4 receptor (H(4)R) has been shown to have preclinical involvement in both inflammatory and pruritic responses. JNJ-39758979 [(R)-4-(3-amino-pyrrolidin-1-yl)-6-isopropyl-pyrimidin-2-ylamine] is a potent and selective H(4)R antagonist with a Ki at the human receptor of 12.5 ± 2.6 nM and greater than 80-fold selectivity over other histamine receptors. The compound also exhibited excellent selectivity versus other targets. JNJ-39758979 showed dose-dependent activity in models of asthma and dermatitis consistent with other H(4)R antagonists. Preclinical toxicity studies of up to 6 months in rats and 9 months in monkeys indicated an excellent safety profile, supporting the clinical testing of the compound. An oral formulation of JNJ-39758979 was studied in a phase 1 human volunteer study to assess safety, pharmacokinetics, and pharmacodynamics. The compound was well tolerated, with the exception of dose-dependent nausea, and no safety issues were noted in the phase 1 study. JNJ-39758979 exhibited good pharmacokinetics upon oral dosing with a plasma half-life of 124-157 hours after a single oral dose. In addition, dose-dependent inhibition of histamine-induced eosinophil shape change was detected, suggesting that the H4R was inhibited in vivo. In conclusion, JNJ-39758979 is a potent and selective H(4)R antagonist that exhibited good preclinical and phase 1 safety in healthy volunteers with evidence of a pharmacodynamics effect in humans.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Cell Shape; Dermatitis, Contact; Double-Blind Method; Eosinophils; Female; Humans; Isothiocyanates; Macaca fascicularis; Male; Ovalbumin; Pyrimidines; Rats; Receptors, G-Protein-Coupled; Receptors, Histamine; Receptors, Histamine H4; Thiazoles

Traditional Chinese medicines (TCM) have been used to treat bronchial asthma for several centuries and a certain degree of clinical benefit has been observed; however, scientific substantiation is lacking. A multicenter, double-blind and placebo-controlled study was therefore conducted to evaluate the clinical efficacy in terms of symptom score, medication score, morning and evening PEFRs, and changes of immunoregulatory function, such as distribution of lymphocyte subsets and in vivo and in vitro production of lymphokines (IFN-gamma and IL-4) and inflammatory mediators (histamine, PGE2 and LTC4). Furthermore, the protective effect of TCM on the late asthmatic reaction (LAR) was evaluated by using asthmatic guinea pigs. Three hundred and three asthmatic children were classified by Chinese doctors, according to a standardized questionnaire designed on the basis of basic logic of Chinese medicine, into three groups of specific constitution (group A, B and C). Group A consisted of 32 herb A-treated patients and 34 placebo-treated; group B, 74 herb B-treated and 64 placebo-treated; and group C, 55 herb C-treated and 44 placebo-treated. The study period was six months. The results were: 1) Both treatment group and placebo group showed an improvement in all clinical parameters, thus demonstrating a placebo effect. However, the improvement was usually greater in the former than the latter, although only the difference in PEFR was significant; 2) Herb A could increase total T cell and decrease B cell; 3) Herb A and B enhanced production of PGE2 but not LTC4, IFN-gamma and IL-4; 4) There was a general tendency for in vivo and in vitro production of histamine to decrease at the end of study in both treatment group and placebo group; however, the decrease was significantly greater in the former than the latter; 5) In asthmatic guinea pigs, 10-day's pretreatment with Chinese herbs could reverse the decrease of sGaw, suppress eosinophilia in bronchoalveolar lavage fluid (BALF), prevent the eosinophil infiltration of airways, increase PGE2 production and decrease LTC4 production in serum and BALF. Thus, traditional Chinese medicines did show a certain degree of clinical efficacy. The decreased production of histamine and LTC4, increased production of PGE2 that were found in both asthmatic children and asthmatic guinea pigs, and prevention of occurrence of LAR by suppressing eosinophil infiltration of airways and preserving airway conductance that were observed in asthm

Topics: Adolescent; Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Child; Dinoprostone; Double-Blind Method; Drug Monitoring; Drugs, Chinese Herbal; Eosinophilia; Eosinophils; Female; Guinea Pigs; Histamine; Humans; Interferon-gamma; Interleukin-4; Kidney; Leukotriene C4; Lymphocyte Subsets; Male; Ovalbumin; Peak Expiratory Flow Rate; Severity of Illness Index; Spleen; Surveys and Questionnaires; Vaccination

Topics: Allergens; Anaphylaxis; Animals; Asthma; Bronchi; Bronchial Provocation Tests; Chromones; Drug Evaluation; Drug Evaluation, Preclinical; Dust; Guinea Pigs; Histamine; Humans; Immunization, Passive; Indomethacin; Male; Ovalbumin; Pyrilamine; SRS-A

Angelica decursiva Franchet & Savatier is a traditional medicinal plant used to treat asthma, cough, headache, pyrexia and thick phlegm in China, Japan and Korea. A. decursiva contains many types of coumarins, which can exert several pharmacological activities including anti-inflammatory and antioxidant properties for treating various diseases such as pneumonitis, atopic dermatitis, diabetes, and Alzheimer's disease.. In this study, we analyzed the components of A. decursiva ethanol extract (ADE) by high performance liquid chromatography (HPLC) and investigated the therapeutic effects of ADE against allergic asthma using lipopolysaccharide (LPS) stimulated RAW264.7 cells and an ovalbumin (OVA)-exposed allergic asthma model. To elucidate the mechanism of action of ADE, we examined the protein expression through network pharmacological analysis.. To establish asthma model, the mice were sensitized on day 0 and 14 via intraperitoneal injection of OVA with aluminum hydroxide. The mice were inhaled with OVA using an ultrasonic nebulizer on day 21, 22 and 23. ADE (50 and 100 mg/kg) was administered to mice by oral gave form day 18-23. On day 24, airway hyperresponsiveness (AHR) was measured using flexivent. On day 25, the mice were sacrificed and collected bronchoalveolar lavage fluids (BALF), serum and lung tissue. In LPS-stimulated RAW264.7 cell, nitric oxide and cytokines were measured. Additionally, expression of nuclear factor erythroid-2-related factor (Nrf2) and suppression of nuclear factor (NF)-κB were detected using double-immunofluorescence.. We detected the five coumarin components which included nodakenin, umbelliferon, (-)-marmesin (=nodakenetin), bergapten, and decursin, in ADE by high performance liquid chromatography. Treatment with ADE decreased the production of nitric oxide, interleukin (IL)-6 and tumor necrosis factor (TNF)-α in LPS-stimulated RAW264.7 cells accompanied by the enhanced expression of nuclear factor erythroid-2-related factor (Nrf2) and suppression of nuclear factor (NF)-κB. In the asthma model, the administration of ADE reduced inflammatory cell count and airway hyperresponsiveness in OVA-exposed animals with decreased levels of IL-4, IL-13, and OVA-specific immunoglobulin E. These results were accompanied by the reduction of pulmonary inflammation and mucus secretion. Furthermore, ADE administration inhibited the expression of NF-κB and matrix metalloproteinase (MMP)-9 in OVA-exposed animals, which was consistent with the results of network pharmacological analysis.. This study demonstrated that ADE effectively attenuated allergic inflammation induced by OVA inhalation through the enhancement of Nrf2 expression and suppression of NF-κB expression. Therefore, ADE may be a potential therapeutic agent for controlling asthma.

Topics: Angelica; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Hypersensitivity; Interleukin-6; Lipopolysaccharides; Lung; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; NF-kappa B; Nitric Oxide; Ovalbumin; Pneumonia

More than 300 million people worldwide suffer from asthma, a chronic respiratory inflammatory disease. Total alkaloids (TA) were extracted from the ethnic medicinal plant Alstonia solaris (L.) R.Br., which is used to treat respiratory diseases. They may be effective drugs for treating asthma, but research is still needed to determine their effectiveness and mechanism in treating asthma.. To further understand TA's role in the treatment of asthma and to support the phase II trial of the drug.. In this study, we investigated the effects of TA in a mouse asthma model produced by Ovalbumin (OVA). H&E and PAS staining were used to observe the histopathological features of lung. airway hyperresponsiveness (AHR) was detected by ventilator; The expression of interleukin (IL)-33, suppression of tumorigenicity 2 (ST2) and E-cadherin in the lungs was evaluated by IHC. The concentrations of Mucin5AC (MUC5AC), eotaxin, IL-4, IL-5, IL-9, IL-13, interferon (IFN)-γ, IL-6, IL-8, IL-17A, IL-33, IL-25, thymic stromal lymphopoietin (TSLP), monocyte chemoattractant protein 1 (MCP-1), leukotriene (LT) B. Administration of TA reduced pulmonary inflammatory symptoms, MUC5AC production in the BALF, and AHR. At the same time, TA inhibited eotaxin production and eosinophil recruitment. Moreover, TA significantly decreased Th2 and Th17 cytokines and increased Th1 cytokines, contributing to restore the balance between Th1 and Th2 and Th17 cytokines. TA may reduce ILC2s numbers by inhibiting IL-33, IL-25, and TSLP levels in BALF and IL-33/ST2 signaling in lung tissue. Finally, TA decreased tIgE, OVA-IgE, and MCP-1 levels and subsequently inhibited mast cell activation and leukotriene release.. These findings imply that TA may be an effective immunoregulatory medication for the management and prevention of asthma.

Topics: Alkaloids; Alstonia; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Immunity, Innate; Immunoglobulin E; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Leukotrienes; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th17 Cells

Eclipta prostrata (L.) L. is a medicinal plant used by many ethnic groups in Brazil to treat respiratory diseases, hepatitis and the bites of venomous animals. A methanolic extract of E. prostrata (MEEP), the major components of which are wedelolactone (WED) and demethylwedelolactone (DMW), exhibited anti-inflammatory activity in acute asthma models but the effects on lung inflammation and the mechanisms of action of MEEP in a chronic asthma model are not known.. To study the effects of MEEP in vivo using a chronic ovalbumin (OVA)-induced allergic asthma model in mice.. The identities of WED and DMW in MEEP were confirmed and the concentrations determined by liquid chromatography and tandem mass spectrometry. Male Balb/c mice were sensitized and challenged with OVA and experimental animals were treated with MEEP (100, 250 or 500 mg/kg) while control animals were treated with dexamethasone (2 mg/kg) or normal saline. Bronchial hyperresponsiveness, total and differential cell counts in bronchoalveolar lavage (BAL), and the production of Th2 cytokines in lung homogenates were assessed. Lung inflammation and mucus production were evaluated by histological analysis while nuclear factor kappa-B (NF-κB) activation was assessed immunohistochemically.. Concentrations of WED and DMW in MEEP were 5.12% and 1.04%, respectively. Treatments with MEEP (250 or 500 mg/kg) significantly decreased bronchial hyperresponsiveness, reduced total cell and eosinophil counts in BAL and IL-4 concentrations in lung homogenate, and inhibited NF-κB activation. Treatment with MEEP at 500 mg/kg reduced the level of IL-5 in lung homogenates but did not decrease IL-13 concentration or mucus production.. MEEP attenuated bronchial hyperresponsiveness and decreased lung and airway inflammation in a chronic asthma model in mice. The mechanism of action involves inhibition of NF-κB activation, most likely associated with the presence of the coumestans WED and DMW. These results support the ethnopharmacological evidence for the use of E. prostrata against asthma and other respiratory inflammatory diseases.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eclipta; Hypersensitivity; Inflammation; Lung; Methanol; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Asthma is a chronic airway inflammatory disease that can lead to several complications caused by bacterial infections. However, recurrent attacks of the disease require long-term use of antibiotics, resulting in lung dysbiosis and poor outcomes. Daqing Formula (DQF) is a well-known herbal medicine in Pharmacopoeia of China, which is widely used for various stimuli-induced lower respiratory diseases, including asthma, bronchitis, and pneumonia. Thus, it has been demonstrated to be a plant-derived broad-spectrum antibiotic for treating and preventing various acute and chronic respiratory diseases.. This study evaluated the efficacy and possible mechanism of DQF on allergic asthma and airway dysbiosis.. The mice were co-challenged with ovalbumin and ampicillin to induce allergic asthma combined with airway dysbacteriosis. The populations of lung microbiota were detected by using 16s DNA sequencing. The levels of asthmatic markers in BALF were detected by ELISA. The levels of Th1/Th2 cytokines in splenic CD4. The results showed that treatment with DQF at 200-800 mg/kg doses significantly reduced the frequency of nasal rubbing and lung inflammation as well as the number of total cells, eosinophils, and macrophages in bronchoalveolar lavage fluid. It decreased the relative abundances of Streptococcus, Cuoriavidus, and Moraxella, increased Akkermansia and Prevotella_6 in lung tissues of asthmatic mice, and inhibited the growth of Staphylococcus aureus, Klebsiella pneumoniae, Streptococcus pneumoniae and their resistant strains in vitro. Furthermore, DQF reduced the levels of eotaxin, TSLP, IL-4, IL-5, IL-25, and IL-33, but enhanced IFN-γ and IL-12 in BALF. It elevated the population of Th1 cells, inhibited eosinophil activation, and downregulated the expressions of p-GSK-3β, p-p65, nuclear β-catenin, and p-STAT3 in the lung tissues of asthmatic mice.. The results revealed that DQF reduced airway inflammation, ameliorated lung dysbiosis, shifted the Th1/Th2 balance, and inhibited eosinophil activation in asthmatic mice, indicating its potential for severe asthma treatment.

Topics: Ampicillin; Animals; Anti-Bacterial Agents; Asthma; Dysbiosis; Glycogen Synthase Kinase 3 beta; Mice; Ovalbumin

Excessive secretion of airway mucus may be an important pathological factor of air pollution-induced acute asthma attacks. Treatment of airway mucus hypersecretion improves asthma aggravated by air pollutants. Qufeng Xuanbi Formula (QFXBF) has been used to treat asthma for more than 30 years. However, whether QFXBF inhibits asthmatic mucus secretion exacerbated by air pollutants has not yet been established.. This study aimed to evaluate the effect of QFXBF on airway mucus secretion and the mechanism of action in an air pollutant benzo[a]pyrene (BaP)-induced mouse model of aggravated asthma.. Ovalbumin (OVA) and BaP co-exposure were used to establish the aggravated asthma model. The average enhanced pause (Penh), serum OVA-specific IgE, and changes in lung histopathology were determined. 16HBE cells exposed to BaP, treatment with QFXBF, arylhydrocarbon receptor (AhR) signal antagonist SR1, reactive oxygen species (ROS) antagonist NAC, or extracellular signal-regulated kinase (ERK1/2) signal antagonist U0126 were established to investigate the effect of QFXBF on BaP-induced mucus secretion and its target. The mRNA and protein expression levels of MUC5AC in the lung tissue and 16HBE cells were examined. We also studied the effect of QFXBF on ROS production. Finally, the protein expression of AhR, phospho-extracellular signal-regulated kinases (p-ERK1/2), and ERK1/2 in 16HBE cells and lung tissues was determined by western blotting.. Administration of QFXBF significantly alleviated the pathological symptoms, including Penh, serum OVA-specific IgE, and changes in lung histopathology in a BaP-induced mouse model of aggravated asthma. QFXBF inhibited MUC5AC expression in asthmatic mice and 16HBE cells exposed to BaP. ROS production, AhR expression, and ERK1/2 phosphorylation were significantly increased in BaP-induced asthmatic mice and 16HBE cells. Signaling pathway inhibitors StemRegenin 1 (SR1), NAC, and U0126 significantly inhibitedBaP-induced MUC5AC expression in 16HBE cells. SR1 reversed Bap-induced ROS production and ERK activation, and NAC inhibited Bap-induced ERK activation. In addition, QFXBF regulated AhR signaling, inhibited ROS production, reversed ERK activation, and downregulated mucus secretion to improve asthma aggravated by air pollutant BaP.. QFXBF can ameliorate mucus secretion in BaP-induced aggravated asthmatic mice and 16HBE cells, and the specific mechanism may be related to the inhibition of the AhR/ROS/ERK signaling pathway.

Topics: Air Pollutants; Animals; Asthma; Benzo(a)pyrene; Disease Models, Animal; Extracellular Signal-Regulated MAP Kinases; Immunoglobulin E; Lung; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Reactive Oxygen Species

Myricetin, a flavonoid isolated from many edible vegetables and fruits, has multiple biological effects, including anti-inflammatory and anti-tumor effects. Myricetin could inhibit mast cell degranulation in vitro, and it reduced the eosinophil content in bronchoalveolar lavage fluid (BALF) of ovalbumin (OVA)-sensitized mice. However, it remains unclear whether myricetin alleviates airway hyperresponsiveness (AHR), airway inflammation, and oxidative stress in asthma. Here, we investigated whether myricetin attenuated AHR, airway inflammation, and eosinophil infiltration in lungs of asthmatic mice. Mice were sensitized with OVA, then injected intraperitoneally with myricetin to investigate anti-inflammatory and antioxidant effects of myricetin. Moreover, we examined its effects on human bronchial epithelial BEAS-2B cells stimulated with TNF-α and IL-4, in vitro. Myricetin effectively mitigated eosinophil infiltration, AHR, and goblet cell hyperplasia in lung, and it reduced Th2 cytokine expression in BALF from asthmatic mice. Myricetin effectively promoted glutathione and superoxide dismutase productions and mitigated malondialdehyde expressions in mice by promoting Nrf2/HO-1 expression. Myricetin also reduced the production of proinflammatory cytokines, eotaxins, and reactive oxygen species in BEAS-2B cells. Myricetin effectively suppressed ICAM-1 expression in inflammatory BEAS-2B cells, which suppressed monocyte cell adherence. These results suggested that myricetin could effectively improve asthma symptoms, mainly through blocking Th2-cell activation, which reduced oxidative stress, AHR, and airway inflammation.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Flavonoids; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress

Melia azedarach L. is a traditional medicinal plant used to control pain, pyrexia, inflammation and bacterial infections that possesses several pharmacological activities, including anti-inflammatory and antioxidant activities. Particularly, the root of M. azedarach was used as expectorant and anti-cough and asthma treatment. Based its properties, M. azedarach is expected to have a potential to treat allergic asthma, chronic inflammatory respiratory disease. However, there is no study on anti-asthmatic effects of M. azedarach and its mechanism of action until now.. We investigated the active ingredient of M. azedarach fruit extract (MAE) using high-performance liquid chromatography (HPLC) and explored the therapeutic effects of MAE on pulmonary inflammation and mucus hypersecretion using a murine model of ovalbumin (OVA) exposed asthma.. The ingredients of MAE were analyzed using HPLC. To develop allergic asthma model, the animals were sensitized (days 1 and 14) and the airway was challenged (from day 21-23) using OVA. MAE was administered by oral gavage once a day from day 18-23 at doses of 30 and 100 mg/kg.. HPLC analysis revealed the presence of toosendanin in MAE. In asthmatic mice, MAE administration effectively suppressed the inflammatory cell counts in bronchoalveolar lavage fluid (BALF) along with a reduction in airway hyperresponsiveness. Moreover, MAE administration inhibited the production of proinflammatory cytokines and immunoglobulin E in BALF and serum of asthmatic mice, respectively. These results were similar to the results of histological examination showing a reduction in pulmonary inflammation and mucus hypersecretion. MAE elevated the expression of nuclear factor erythroid 2-related factor 2, heme oxygenase-1, and superoxide dismutase 2, which in turn resulted in the suppression of matrix metallopeptidase-9 expression in lung tissue of asthmatic mice.. Altogether, MAE successfully inhibited allergic asthma in OVA-exposed mice. Thus, MAE could be a potential therapeutic remedy for treating allergic asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Melia azedarach; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Pneumonia

Icariin, a flavonoid glycoside isolated from

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Corticosterone; Cytokines; Depression; Disease Models, Animal; DNA; Flavonoids; Glucocorticoids; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Receptors, Glucocorticoid

Asthma is a common respiratory disease that has no definitive treatment at now. Immune response shifting from T helper (Th)1 to the Th2 is a main problem in asthma, and immunomodulation can help to control asthma. IL-35 and mesenchymal stem cells (MSCs) have regulatory effect on the immune system and may have the ability to control asthma pathology. After culturing MSCs, expression vector of IL-35 (pUNO1-mIL35elasti) was transduced to the MSCs, and then, asthmatic mice were treated with MSCs, MSCs-vector, MSCs-vector-IL-35, and no treatment. Airway hyperresponsiveness (AHR), levels of the cytokines, total and ovalbumin (OVA) specific immunoglobulin (Ig)E, LTB4, and LTC4 were measured. Lung tissue histopathology was also done. MSCs were successfully transduced by pUNO1-mIL35elasti vector, and IL-35 was produced in transduced cells. AHR, levels of the cytokines, IgEs, LTs, goblet cell hyperplasia, mucus secretion, peribronchial, and perivascular inflammation were controlled by MSCs therapy. In MSCs-IL-35 group, these controls were stronger than MSCs without IL-35 group. MSCs had strong effect on control of asthma. Transfected MSCs by expressing IL-35 gene could significantly better control allergic asthma symptoms than MSCs without IL-35. In the future, identification of the IL-35 mechanism of action would be useful to improve cytokine-cell based therapies.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Immunity; Immunoglobulin E; Immunomodulation; Interleukins; Lung; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Ovalbumin

Radix Paeoniae Alba (RPA), a traditional Chinese medicine, has been used frequently in the treatment of asthma. Previous studies demonstrated the dichloromethane fraction of Stir-Frying RPA (FDCM) enhanced the effect of anti-allergic asthma compared with the dichloromethane fraction of RPA (DCM).. The significant increasing of Paeoniflorin (PF), ethyl gallate (EG), 1,2,3,4,6-pentagalloylglucose (PGG) had been observed in FDCM. This study aimed to investigate the effects and mechanisms of these compounds from FDCM in ovalbumin (OVA)-induced allergic asthma mouse model.. The significant difference contents compounds fraction (FB-40) and other fractions in FDCM were enriched by Medium Pressure Liquid Chromatography (MPLC). The pharmacodynamics was verified among all fractions in OVA-induced allergic asthma mice. Moreover, the drug dose dependence of FB-40 (0.42 mg/kg, 0.21 mg/kg, and 0.07 mg/kg), which were the most active fraction from FDCM for anti-allergic asthma, was explored. The expression of IL-6, p-STAT3, and STAT3 was analyzed by Western blot analysis. In addition, the main components of FB-40 were identified by UPLC with standards. Finally, the anti-inflammatory effects of the main components from FB-40 were detected by LPS-stimulated BEAS-2B cells using an Elisa assay.. The results showed that FB-40 was the most active fraction from FDCM, which could significantly improve the lung tissue pathological condition, and decrease the number of inflammatory cells in bronchoalveolar lavage fluid (BALF). It had greater pharmacological activity than its main component PF. FB-40 also showed dose dependence and regulated the IL-6/STAT3 signaling pathway in allergic asthma mice. Besides, PF, Albiflorin (AF), PGG, EG, and 1,2,3,6-Tetra-O-galloyl-β-D-glucose (TGG) from FB-40 were identified by UPLC with the standard. At last, in the LPS-induced BEAS-2B cell experiments, EG, PGG, 1,2,3,6-Tetra-O-galloyl-β-D-glucose (TGG) showed stronger inhibiting activities of cytokine than the monoterpenoid glycosides (PF and AF).. The research proved that FB-40 was an active fraction in FDCM, which regulates IL-6/STAT3 Signaling Pathway to ameliorate allergic asthma. Gallic acids including TGG and PGG, and EG also play a role in the treatment of allergic asthma in FB-40.

Topics: Animals; Anti-Allergic Agents; Asthma; Bronchoalveolar Lavage Fluid; Glucose; Interleukin-6; Lipopolysaccharides; Methylene Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Signal Transduction

To investigate the therapeutic and metabolic regulatory effects and the underlying mechanism of HPMHD in asthmatic rats.. The asthmatic rats were administered with the corresponding HPMHD (at dosages of 5.54, 11.07, 22.14 mg/kg). Then inflammatory cells in peripheral blood and bronchoalveolar lavage fluid (BALF) were counted, the levels of interleukin (IL)-4 and IL-13 in BALF were measured, and the changes in enhanced pause (Penh) and pathological damage of lung tissues were also detected to evaluate the protective effects of HPMHD. The serum metabolic profile of HPMHD in asthmatic rats was explored using Ultra-High-Performance Liquid Chromatography-mass spectrometer (UHPLC-MS), and the regulatory effects on TRPV1 and TJs of HPMHD in asthmatic rats were detected by Western blotting analysis. In vitro, 16HBE cells were stimulated with IL-4 plus SO. HPMHD significantly attenuated the airway inflammation of asthmatic rats, and reduced the levels of inflammatory cells in peripheral blood and BALF as well as the levels of IL-4 plus IL-13 in BALF. In addition, the airway hyperresponsiveness and lung pathological damage were alleviated. Serum metabolomic analysis showed that 31 metabolites were differentially expressed among the normal saline-, model-, and HPMHD-treated rats. Pathway enrichment analysis showed that the metabolites were involved in 45 pathways, among which, TJs regulation-relevant pathway was associated with the Ca. The current study indicates that HPMHD alleviates rat asthma and participates in the regulation of serum metabolism. The anti-asthma effects of HPMHD might be related to the protection of TJs by inhibiting the intracellular Ca

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Interleukin-13; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Rats; TRPV Cation Channels

The present study was intended to explore the valuable effects of triptonide on inflammation, asthmatic, and nociceptive. Triptonide possesses numerous beneficial effects extensively managed in the treatment of inflammation disease condition. Initially, triptonide showed anti-inflammation properties over lipopolysaccharide-induced RAW 264.7 cells. Hence, the present study was directed to explore the protecting efficacy of triptonide in ovalbumin (OVA)-induced asthma in mice. Asthma was induced intraperitoneally administration (200μL) in female BALB/c mice with suspension which has ovalbumin (100 μg/mL) and aluminum hydroxide (10 mg/mL). Triptonide (30 mg/kg) over OVA-induced experimental animals altered lung mass, nitric oxide, myeloperoxidase, immunoglobulin E status, interleukins (4, 5, and 13) inflammatory cytokines status, and histological modifications. Animals were also managed with the standard drug dexamethasone (50 mg/kg) followed by the asthma induction, which is also efficient over OVA-induced experimental animals. The nociception was provoked in male Swiss mice by various chemicals (acetic acid, capsaicin, and glutamate). The animals were administered with triptonide (5, 10, and 15 mg/kg) and separate standard drugs like diclofenac sodium (10 mg/kg) and morphine (5 mg/kg) over chemical-induced nociceptive animals. The present outcome evidently established that the triptonide considerably reduced the various chemical-induced nociception in mice (Fig. 7A, B, and C). Ultimately, the present work explored the evident powerful anti-inflammation, antinociceptive, and anti-asthma properties of a diterpenoid, triptonide experimental animal models. And it is recommended that triptonide is an excellent compound in the management of asthma and its related diseases.

Topics: Allergens; Analgesics; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Diterpenes; Female; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin

Rhinovirus infection frequently causes COPD and asthma exacerbations. Impaired anti-viral signaling and reduced viral clearance have both been seen in sick bronchial epithelium, potentially increasing exacerbations. Polyinosinic:polycytidylic acid (Poly(I:C)), a Toll-like receptor-3 (TLR3) ligand, has been shown to cause a viral exacerbation of severe asthma by detecting double-stranded RNA (dsRNA). The purpose of this work was to determine the effect of a TLR3/dsRNA complex inhibitor-Calbiochem drug in the prevention of Poly(I:C)-induced airway inflammation following TLR3 activation and to uncover a potential pathway for the cure of asthma through TLR3 inhibition. Mice were sensitized with Poly(I:C) as an asthma model before being challenged by PBS and ovalbumin (OVA) chemicals. The mice were administered a TLR3/dsRNA complex inhibitor. Throughout the trial, the mice's body weight was measured after each dosage. Biochemical methods are used to analyze the protein as well as enzyme composition in airway tissues. BALF specimens are stained using Giemsa to identify inflammatory cells and lung histopathology to determine morphological abnormalities in lung tissues. By using the ELISA approach, cytokine levels such as TNF-α, IL-13, IL-6, IL-5, and IgE antibody expression in lung tissue and blood serum were assessed. TLR3/dsRNA complex inhibitor drug significantly lowered the number of cells in BALF and also on Giemsa staining slides. It also downregulated the level of TNF-α and IL-6 in contrast to OVA and Poly(I:C) administered in animals. A TLR3/dsRNA complex inhibitor decreased the fraction of oxidative stress markers (MDA, GSH, GPx, and CAT) in lung tissues while keeping the mice's body weight constant during the treatment period. By decreasing alveoli, bronchial narrowing, smooth muscle hypertrophy, and granulocyte levels, the TLR3/dsRNA complex blocker significantly reduced the histopathological damage caused by OVA and Poly(I:C) compounds. In an animal model utilizing ovalbumin, TLR3/dsRNA complex inhibitors similarly reduced the bronchial damage produced by Poly(I:C). A novel TLR3/dsRNA complex inhibitor is expected to be employed in clinical studies since it suppresses airway inflammation without inducing antiviral approach resistance.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Interleukin-6; Lung; Mice; Ovalbumin; Poly I-C; RNA, Double-Stranded; Toll-Like Receptor 3; Tumor Necrosis Factor-alpha

Early-life exposure to certain environmental bacteria including Acinetobacter lwoffii (AL) has been implicated in protection from chronic inflammatory diseases including asthma later in life. However, the underlying mechanisms at the immune-microbe interface remain largely unknown.. The effects of repeated intranasal AL exposure on local and systemic innate immune responses were investigated in wild-type and Il6. The asthma preventive effect of AL was confirmed in the lung. Repeated intranasal AL administration triggered a proinflammatory immune response particularly characterized by elevated levels of IL-6, and consequently, IL-6 induced IL-10 production in CD4. These experiments provide a novel mechanism of Acinetobacter lwoffii-induced asthma protection operating through IL-6-mediated epigenetic activation of IL-10 production and with associated effects on the intestinal microbiome.

Topics: Administration, Intranasal; Animals; Asthma; Disease Models, Animal; Inflammation; Interleukin-10; Interleukin-6; Lung; Mice; Mice, Inbred BALB C; Microbiota; Ovalbumin

Leonurine (Leo) is a natural alkaloid extracted from Herba leonuri, which has many biological activities. However, whether leonurine has a protective effect on asthma remains unknown. The purpose of this study was to investigate the protective effect of leonurine on asthma. We evaluated its therapeutic effect and related signal transduction in LPS-induced RAW264.7 cells and OVA-induced asthmatic mice. In addition, we used network pharmacology, molecular docking and molecular dynamics simulation to verify the experimental results. In LPS-induced RAW 264.7 cells, leonurine significantly reduced the production of TNF-α and IL-6, andinhibited the activation of p38 MAPK/NF-κB signaling pathway. In OVA-induced asthmatic mice, leonurine decreased the number of inflammatory cells in the bronchoalveolar lavage fluid (BALF), particularly neutrophils and eosinophils. Leonurine also reduced the contents of IL-4, IL-5, IL-13 in the BALF and OVA-IgE in the serum. Leonurine remarkly improved OVA-induced inflammatory cell infiltration and significantly inhibited mucus overproduction. In addition, leonurine inhibited the activation of p38 MAPK/NF-κB signaling pathway in the lung tissues of asthmatic mice. Network pharmacology suggested that p38 MAPKα was a potential target of leonurine in the treatment of asthma. Molecular docking and molecular dynamics simulations indicated that leonurine could stably bind to p38 MAPKα protein. In summary, leonurine attenuated asthma by regulating p38 MAPK/NF-κB signaling pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Molecular Docking Simulation; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Signal Transduction

The current study was designed to investigate the potential anti-inflammatory and antioxidant effects of fingolimod against Ovalbumin (Ova)-induced allergic airway inflammation compared to dexamethasone. Fingolimod was given (0.5 mg/kg/day, p.o.) for sensitized mice 1 h before Ova challenge from Days 19 to 24. Fingolimod significantly inhibited Ova-induced elevation of inflammatory cells and eosinophils numbers in bronchoalveolar lavage fluid (BALF) and reduced concentrations of immunoglobulin E in serum and of sphingosine-1-phosphate, interleukin (IL)-4, and IL-13 in BALF. Fingolimod inhibited microvascular leakage and edema as reflected by the decreased lung/body weight index. These findings were supported by histopathological examination results showing that fingolimod substantially decreased perivascular edema and inflammatory cell infiltration. Fingolimod also attenuated Ova-induced oxidative stress as evidenced by decreased malondialdehyde concentration along with increasing concentrations of reduced glutathione and superoxide dismutase in lung tissues. Fingolimod also significantly decreased monocyte chemoattractant protein-1 (MCP-1), p-ERK, and p-P38 in lung tissues of Ova-challenged mice. In conclusion, the current study demonstrated the anti-inflammatory and antioxidant effects of fingolimod in allergic airway inflammation that might be associated with the downregulation of mitogen activated kinases signaling to decrease T helper 2 cytokine secretion (IL-4 and IL-13) and MCP-1 expression, along with the inhibition of oxidative stress.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Fingolimod Hydrochloride; Inflammation; Interleukin-13; Lung; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Ovalbumin

Air pollution and environmental issues significantly impact life, resulting in the emergence and exacerbation of allergic asthma and other chronic respiratory infections. The main objective of this study is to suppress allergic asthma by TAK-242 from lipopolysaccharide-induced airway inflammation primarily stimulating toll-like receptor-4, and also to determine the potential mechanism of asthma eradication. The TAK-242 anti-allergic action was assured through the ovalbumin murine model of asthma via bronchial hyperresponsiveness and inflammation of the respiration tract in a pre-existing allergic inflammation paradigm. Swiss albino mice were sensitized and then challenged by ovalbumin and lipopolysaccharide for 5 days straight. TAK-242 reaction was assessed by inflammatory cytokines, and inflammatory cell count was determined from blood serum and bronchoalveolar lavage fluid, as well as group-wise regular weight assessments. After ovalbumin, lipopolysaccharide infusion, toll-like receptor-4 agonists caused a substantial increase in airway hyperresponsiveness, specific cellular inflammation, histological alterations, and immune mediator synthesis, as well as dose-related body-weight variations. A decrease in lipopolysaccharide-induced leukocyte count and Th1/Th17 related cytokines, TNF-α, and IL-6 expression through the ELISA study was particularly noticeable. Finally in treated groups, TAK-242, a TLR4/MD2 complex inhibitor, reduced airway inflammation and histopathological changes, cytokine expression, and body-weight management. TAK-242 has been found in an ovalbumin allergic asthma model to be a potential inhibitor of lipopolysaccharide-induced respiratory infection.

Topics: Animals; Asthma; Bacterial Load; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Toll-Like Receptor 4

In most asthma patients, symptoms are controlled by treatment with glucocorticoid, but long-term or high-dose use can produce adverse effects. Therefore, it is crucial to find new therapeutic strategies. β-sitosterol could suppress type Ⅱ inflammation in ovalbumin (OVA)-induced mice, but its mechanisms have remained unclear.. A binding activity of β-sitosterol with glucocorticoid receptor (GR) was analyzed by molecular docking. Human bronchial epithelial cells (BEAS-2B) and human bronchial smooth muscle cells (HBSMC) were treated with different concentrations (0, 1, 5, 10, 20, and 50 μg/mL) of β-sitosterol for suitable concentration selection. In transforming growth factor (TGF)-β1 treated BEAS-2B and HBSMC, cells were treated with 20 μg/mL β-sitosterol or dexamethasone (Dex) to analyze its possible mechanism. In OVA-induced mice, 2.5 mg/kg β-sitosterol or Dex administration was performed to analyze the therapeutic mechanism of β-sitosterol. A GR antagonist RU486 was used to confirm the mechanism of β-sitosterol in the treatment of asthma.. A good binding of β-sitosterol to GR (score = -8.2 kcal/mol) was found, and the GR expression was upregulated with β-sitosterol dose increase in BEAS-2B and HBSMC. Interleukin (IL)-25 and IL-33 secretion was significantly decreased by β-sitosterol in the TGF-β1-induced BEAS-2B, and the levels of collagen 1A and α-smooth muscle actin (SMA) were reduced in the TGF-β1-induced HBSMC. In the OVA-challenged mice, β-sitosterol treatment improved airway inflammation and remodeling through suppressing type Ⅱ immune response and collagen deposition. The therapeutic effects of β-sitosterol were similar to Dex treatment in vitro and in vivo. RU486 treatment clearly hampered the therapeutic effects of β-sitosterol in the TGF-β1-induced cells and OVA-induced mice.. This study identified that β-sitosterol binds GR to perform its functions in asthma treatment. β-sitosterol represent a potential therapeutic drug for allergic asthma.

Topics: Airway Remodeling; Animals; Asthma; Collagen; Disease Models, Animal; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Mifepristone; Molecular Docking Simulation; Ovalbumin; Receptors, Glucocorticoid; Sitosterols; Transforming Growth Factor beta1

Although the T helper 2 (Th2) subset is a critical player in the humoral immune response to extracellular parasites and suppression of Th1-mediated inflammation, Th2 cells have been implicated in allergic inflammatory diseases such as asthma, allergic rhinitis, and atopic dermatitis. GATA binding protein 3 (GATA3) is a primary transcription factor that mediates Th2 differentiation and secretion of Th2 cytokines, including IL-4, IL-5, and IL-13. Here, a nucleus-deliverable form of GATA3-transcription modulation domain (TMD) (ndG3-TMD) was generated using Hph-1 human protein transduction domain (PTD) to modulate the transcriptional function of endogenous GATA3 without genetic manipulation. ndG3-TMD was shown to be efficiently delivered into the cell nucleus quickly without affecting cell viability or intracellular signaling events for T cell activation. ndG3-TMD exhibited a specific inhibitory function for the endogenous GATA3-mediated transcription, such as Th2 cell differentiation and Th2-type cytokine production. Intranasal administration of ndG3-TMD significantly alleviated airway hyperresponsiveness, infiltration of immune cells, and serum IgE level in an OVA-induced mouse model of asthma. Also, Th2 cytokine secretion by the splenocytes isolated from the ndG3-TMD-treated mice substantially decreased. Our results suggest that ndG3-TMD can be a new therapeutic reagent to suppress Th2-mediated allergic diseases through intranasal delivery.

Topics: Administration, Intranasal; Animals; Asthma; Cell Nucleus; Cytokines; Disease Models, Animal; GATA3 Transcription Factor; Humans; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; Th2 Cells

Salidroside (SAL) is a natural component derived from

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Hormones; Lung; Metalloproteases; Mice; Mice, Inbred BALB C; Ovalbumin; Pyrimidines; Steroids

Clarifying the pathogenesis of asthma and/or identifying the specific pathway underlying oral asthma tolerance (OT) would be of great significance. In our previous study, promyelocytic leukemia zinc finger (PLZF), which reportedly regulates memory phenotypes, was found to promote ovalbumin (OVA)-induced OT. Therefore, this study aimed to explore the regulatory effects of PLZF on memory phenotypes in asthma and OT mouse models. We found that Zbtb16 (encoding PLZF) and PLZF

Topics: Animals; Asthma; Hyaluronan Receptors; Lung; Lymphocytes; Mice; Ovalbumin; Phenotype; Promyelocytic Leukemia Zinc Finger Protein; Respiratory Syncytial Virus Infections

Allergic asthma is a chronic inflammatory disease primarily mediated by Th2 immune mechanisms. Exposure to antibiotics during early life is associated with an increased risk of allergic asthma, although the exact mechanism is not fully understood. In this study, mice were randomly divided into a normal saline control group (NS group), an OVA-induced asthma group (OVA group), a vancomycin treatment control group (VAN.NS group), and a vancomycin treatment the OVA-induced asthma group (VAN.OVA group). The results showed that vancomycin altered dominant species in experimental mice. The phylum level histogram showed that Bacteroides abundance was increased, and Firmicutes abundance was decreased in the OVA group. Airway inflammation and airway hyperresponsiveness (AHR) were aggravated in the vancomycin-exposed group. Enzyme-linked immunosorbent assay (ELISA) showed that the serum levels of IL-5, IL-13, and IL-33 in the OVA group were higher than those in the NS group, especially in the VAN.OVA group. The expression of GATA binding protein-3(GATA3) and retinoid acid receptor-related orphan receptor alpha (RORa) increased in the OVA group, even more so in the VAN.OVA group. Group 2 innate lymphoid cells (ILC2s) in the lung detected by flow cytometry was increased in OVA mice more than those in control mice, with a more remarkable increase in the VAN.OVA. Our results demonstrated that vancomycin used in early life could alter the intestinal microecology of mice, which, in turn, aggravates airway inflammation and upregulate type 2 innate lymphocytes.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Gastrointestinal Microbiome; Immunity, Innate; Inflammation; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Vancomycin

Bronchial asthma, a non-communicable chronic respiratory disease, affects people of all ages. An important pathological feature of bronchial asthma is airway remodeling. Hyssopus cuspidatus Boriss. has been used to treat bronchial asthma for over 100 years in Uygur medicine. The ethanol extract of Hyssopus cuspidatus Boriss.(JAX2) can improve airway inflammation in asthma. However, the anti-asthmatic airway-remodeling effect of JAX2 is unclear.. The current study investigated the anti-airway remodeling effect of JAX2 and elucidated its mechanism of action.. The present study established an ovalbumin-induced mouse model of asthma and platelet-derived growth factor-BB-induced human airway smooth muscle cells (hASMCs) proliferation model, with dexamethasone (DEX) and feining tablets (FNP) designated as positive control drugs. Pathological changes in lung tissues were observed using hematoxylin and eosin staining. Interleukin (IL)-5, IL-10, IL-13, and IL-33 levels in the bronchoalveolar lavage fluid (BALF) and serum of mice were determined using enzyme-linked immunosorbent assay (ELISA). Changes in the expression and distribution of TGF-β1, p-ERK1/2, Smad2/3, and p-Smad3 in lung tissues were determined using immunohistochemistry. Western blotting (WB) was used to determine the protein levels of p-ERK1/2 in lung tissues and cells. MTS assay was used to determine the effects of JAX2 on cell proliferation. IL-5, IL-10, IL-13, MMP-2, and MMP-9 levels in the cell supernatant were determined using ELISA. HASMCs migration was observed using the scratch and transwell methods. The effect of JAX2 on the hASMCs cycle was determined using flow cytometry.. JAX2 significantly improved the pathological status of lung tissues in asthmatic mice. It could also significantly reduce IL-5, IL-13, and IL-33 levels in the BALF and serum of asthmatic mice in a dose-dependent manner and significantly increase IL-10 levels. TGF-β1, p-ERK1/2, Smad2/3, and p-Smad3 expression in lung tissues were decreased in a dose-dependent manner. The protein level of p-ERK1/2 in lung tissues was also reduced. JAX2 could significantly inhibit the proliferation and migration of PDGF-BB-induced hASMCs. IL-5, IL-13, MMP-9, and MMP-2 levels decreased significantly, and IL-10 levels increased significantly in a dose-dependent manner in the cell supernatant. JAX2 could block hASMCs in the G0/G1 phase, thereby inhibiting cell proliferation. p-ERK1/2 protein levels were found to decrease in a dose-dependent manner.. JAX2 significantly inhibits airway remodeling in asthma. Its mechanism of action may be inhibiting the proliferation and migration of hASMCs, releasing inflammatory factors and metalloproteinases, activating the ERK1/2 signal pathway, and promoting the secretion of anti-inflammatory factors.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Proliferation; Disease Models, Animal; Humans; Hyssopus Plant; Interleukin-10; Interleukin-13; Interleukin-33; Interleukin-5; Lung; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Transforming Growth Factor beta1

Allergic asthma is a heterogeneous disease involving a variety of inflammatory cells. Immune imbalance or changes in the immune microenvironment are the essential causes that promote inflammation in allergic asthma. Tetraspanin CD81 can be used as a platform for receptor clustering and signal transmission owing to its special transmembrane structure and is known to participate in the physiological processes of cell proliferation, differentiation, adhesion, and migration. Previous studies have shown that CD81-targeting peptidomimetics exhibit anti-allergic lung inflammation. However, due to the low metabolic stability of peptide drugs, their druggability is limited. Here, we aimed to generate a metabolically stable anti-CD81 peptide, evaluate its anti-inflammatory action and establish its mechanism of action. Based on previous reports, we applied retro-inverse peptide modification to obtain a new compound, PD00 (NH2-D-Gly-D-Ser-D-Thr-D-Tyr-D-Thr-D-Gln-D-Gly-COOH), with high metabolic stability. Enhanced ultraperformance liquid chromatography-tandem mass spectrometry was used to investigate the in vitro and in vivo metabolic stabilities of PD00. The affinities of PD00 and CD81 were studied using molecular docking and surface plasmon resonance techniques. An ovalbumin (OVA)-induced asthma model was used to evaluate the effects of PD00 in vivo. Mice were treated with different concentrations of PD00 (175 and 350 μg/kg) for 10 days. Airway hyperresponsiveness (AHR) to acetyl-β-methacholine (Mch), inflammatory cell counts in the bronchoalveolar lavage fluid, and serum OVA-specific IgE levels were detected in the mice at the end of the experiment. Lung tissues were collected for haematoxylin and eosin staining, untargeted metabolomic analysis, and single-cell transcriptome sequencing. PD00 has a high affinity for CD81; therefore, administration of PD00 markedly ameliorated AHR and airway inflammation in mice after OVA sensitisation and exposure. Serum OVA-specific IgE levels decreased considerably. In addition, PD00 treatment increased glycerophospholipid and purine metabolism in immune cells. Collectively, PD00 may regulate the glycerophospholipid and purine metabolism pathways to ameliorate the pathophysiological features of asthma. These findings suggest that PD00 is a potential compound for the treatment of asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Immunoglobulin E; Inflammation; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Molecular Docking Simulation; Ovalbumin; Purines

Cough variant asthma (CVA) is a chronic inflammatory disease characterized by cough as the main symptom. Suhuang antitussive capsule (Suhuang), one of traditional Chinese patent medicines, mainly treats CVA clinically. Previous studies have shown that Suhuang significantly improved CVA, post-infectious cough (PIC), sputum obstruction and airway remodeling. However, the effect of Suhuang on ovalbumin-induced (OVA-induced) metabolic abnormalities in CVA is unknown.. This study aimed to identify potential metabolites associated with efficacy of Suhuang in the treatment of CVA, and determined how Suhuang regulates metabolites, and differential metabolites reduce inflammation and oxidative stress.. Rats were given 1 mg OVA/100 mg aluminum hydroxide in the 1st and 7th days by intraperitoneal injection and challenged by atomizing inhalation of 1% OVA saline solution after two weeks to establish the CVA model. Rats were intragastrically (i.g.) administrated with Suhuang at 1.4 g/kg and β-hydroxybutyric acid (β-HB) were given with different concentrations (87.5 and 175 mg/kg/day) by intraperitoneal injection for 2 weeks. After 26 days, GC-MS-based metabolomic approach was applied to observe metabolic changes and search differential metabolites. The number of coughs, coughs latencies, enzyme-linked immunosorbent assay (ELISA), histological analysis and quantitative-polymerase chain reaction (Q-PCR) were used to investigate the effects of Suhuang. Then β-HB on CVA rats, NLRP3 inflammasome and GSK3β/AMPK/Nrf2 signalling pathway were detected by western blotting.. The results showed that Suhuang treatment significantly enhanced the serum level of β-HB. Interestingly, exposure to exogenous β-HB was also protective against OVA-induced CVA. β-HB significantly reduced the number of coughs and lengthened coughs latencies, improved lung injury, reduced the secretion of various cytokines, and directly inhibited the NLRP3 inflammasome. In addition, β-HB increased the nuclear accumulation of Nrf2 by activating the GSK3β/AMPK signaling axis, and then inactivating the NF-κB signaling pathway, effectively protecting OVA-induced CVA from oxidative stress and inflammation.. The results of this study shows that β-HB can reduce inflammation and oxidative stress, the increased production of β-HB in serum might be the crucial factor for Suhuang to exert its effect in the treatment of CVA.

Topics: 3-Hydroxybutyric Acid; AMP-Activated Protein Kinases; Animals; Antitussive Agents; Asthma; Cough; Glycogen Synthase Kinase 3 beta; Inflammasomes; Inflammation; NF-E2-Related Factor 2; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Rats

Elongation of very-long-chain fatty acids protein 6 (ELOVL6), an enzyme regulating elongation of saturated and monounsaturated fatty acids with C12 to C16 to those with C18, has been recently indicated to affect various immune and inflammatory responses; however, the precise process by which ELOVL6-related lipid dysregulation affects allergic airway inflammation is unclear.. This study sought to evaluate the biological roles of ELOVL6 in allergic airway responses and investigate whether regulating lipid composition in the airways could be an alternative treatment for asthma.. Expressions of ELOVL6 and other isoforms were examined in the airways of patients who are severely asthmatic and in mouse models of asthma. Wild-type and ELOVL6-deficient (Elovl6. ELOVL6 expression was downregulated in the bronchial epithelium of patients who are severely asthmatic compared with controls. In asthmatic mice, ELOVL6 deficiency led to enhanced airway inflammation in which lymphocyte egress from lymph nodes was increased, and both type 2 and non-type 2 immune responses were upregulated. Lipidomic profiling revealed that the levels of palmitic acid, ceramides, and sphingosine-1-phosphate were higher in the lungs of ovalbumin-immunized Elovl6. This study illustrates a crucial role for ELOVL6 in controlling allergic airway inflammation via regulation of fatty acid composition and ceramide-sphingosine-1-phosphate biosynthesis and indicates that ELOVL6 may be a novel therapeutic target for asthma.

Topics: Animals; Asthma; Ceramides; Disease Models, Animal; Inflammation; Mice; Ovalbumin

Epidemiological surveys indicate that intrauterine inflammation increases the risk of asthma in offspring. However, the underlying mechanisms remain largely unknown. Previous studies in BALB/c and C57BL/6 mice showed that prenatal exposure to endotoxins prevented allergic airway inflammation in offspring, which is inconsistent with most clinical observations. In this study, we found that prenatal LPS exposure increased airway resistance and total exfoliated cell counts, eosinophils, and IL-4 concentrations in BAL fluid of ovalbumin-sensitized Institute of Cancer Research (ICR) mice. Importantly, long noncoding RNA (lncRNA) NONMMUT033452.2 was upregulated in the lungs of LPS-exposed ICR offspring. Fluorescence

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Humans; In Situ Hybridization, Fluorescence; Inflammation; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Peptide Elongation Factor 1; Pregnancy; Protein Binding; RNA, Long Noncoding

Acupuncture has been frequently used in China for the treatment asthma for thousands of years. Ferroptosis was recently revealed to be involved in several pathological conditions including asthma. However, the detailed links between ferroptosis and airway inflammation in asthma, as well as the detailed regulation of acupuncture on these disorders remains unclear. Our results demonstrated that the non-haem Fe

Topics: Acupuncture Therapy; Animals; Anti-Asthmatic Agents; Arachidonate 15-Lipoxygenase; Asthma; Coenzyme A Ligases; Disease Models, Animal; Ferroptosis; Glutathione Disulfide; Inflammation; Interleukin-4; Mice; Ovalbumin; Pneumonia

Allergic asthma is associated with airflow obstruction and hyper-responsiveness that arises from airway inflammation and remodeling. Cell therapy with mesenchymal stem cells (MSC) has been shown to attenuate inflammation in asthma models, and similar effects have recently been observed using extracellular vesicles (EV) obtained from these cells. Biologically functional vesicles can also be artificially generated from MSC by extruding cells through membranes to produce EV-mimetic nanovesicles (NV). In this study, we aimed to determine the effects of different MSC-derived vesicles in a murine model of allergic airway inflammation.. EV were obtained through sequential centrifugation of serum-free media conditioned by human bone marrow MSC for 24 h. NV were produced through serial extrusion of the whole cells through filters. Both types of vesicles underwent density gradient purification and were quantified through nanoparticle tracking analysis. C57BL/6 mice were sensitized to ovalbumin (OVA, 8 µg), and then randomly divided into the OVA group (intranasally exposed to 100 µg OVA for 5 days) and control group (exposed to PBS). The mice were then further divided into groups that received 2 × 10. Administration of EV and NV reduced cellularity and eosinophilia in bronchoalveolar lavage (BAL) fluid in OVA-sensitized and OVA-exposed mice. In addition, NV treatment resulted in decreased numbers of inflammatory cells within the lung tissue, and this was associated with lower levels of Eotaxin-2 in both BAL fluid and lung tissue. Furthermore, both intranasal and systemic administration of NV were effective in reducing inflammatory cells; however, systemic delivery resulted in a greater reduction of eosinophilia in the lung tissue.. Taken together, our results indicate that MSC-derived NV significantly reduce OVA-induced allergic airway inflammation to a level comparable to EV. Thus, cell-derived NV may be a novel EV-mimetic therapeutic candidate for treating allergic diseases such as asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophilia; Humans; Immunoglobulin E; Inflammation; Lung; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin

Asthma is a chronic airway disease. Allergic reactions and T helper (h)2 immune response play a key role in asthma occurrence. Cell therapy can control inflammation and remodeling responses in allergic asthma, and cytokines can change this effect. Therefore, in this study, the effect of treated cell therapy with IL-2 to control allergic asthma was studied. Bone marrow cells were extracted and co-cultured with IL-2 and the cells were used via intra-tracheal administration in allergic asthma mice. Levels of IL-4, IL-5, IL-13, Leukotriene B4 and C4, and remodeling factors were measured. At least, a histopathology test of lung tissue was done. Type2 cytokines, leukotrienes, remodeling factors, mucus secretion, goblet cell hyperplasia, peri-bronchial and peri-vascular inflammation were significantly (p˂0.05) decreased by treating with bone marrow-derived mononuclear cells (BMDMCs) and IL-2-BMDMCs. Treatment with IL-2-BMDMCs could significantly decrease IL-13, transforming growth factor (TGF)-β, HP levels, and mucus secretion (p˂0.05) compared to BMDMCs treatment. In this study, BMDMCs and IL-2-BMDMCs therapy could decrease inflammation, allergic, and remodeling factors in allergic asthma. Cell therapy with BMDMCs had a strong and notable effect on the control of allergic asthma pathophysiology when co-cultured and used with IL-2.

Topics: Animals; Asthma; Bone Marrow; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Inflammation; Interleukin-13; Interleukin-2; Leukocytes, Mononuclear; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Transforming Growth Factor beta

Asthma is a hackneyed chronic inflammatory disease of the airway. Chryseriol (CSR) is a kind of flavonoid, and has the effect of bronchiectasis, indicating its potential application for treating respiratory diseases. However, the functions of CSR in asthma have not been reported till now.. The histopathologic changes of the lung tissues were assessed by hematoxylin and eosin staining. The cell apoptosis was identified through terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling assay. Total numbers of eosinophils, neutrophils, and macrophages were assessed under microscope. The levels of interleukin (IL)-1β, IL-4, IL-5, and IL-13 were detected by enzyme-linked-immunosorbent serologic assay. The airway hyper-responsiveness (AHR) was evaluated by the whole body plethysmography. The levels of methane dicarboxylic aldehyde, superoxide dismutase, glutathione S-transferase, and glutathione in lung homogenates were confirmed by using corresponding commercial kits. The protein expressions were examined by Western blot analysis.. The ovalbumin (OVA) was utilized to establish asthma mouse model. At first, it was revealed that CSR treatment reduced lung injury in OVA-stimulated mice. Moreover, cell apoptosis was enhanced after OVA stimulation but was attenuated by CSR treatment. In addition, CSR treatment decreased the infiltration of inflammatory cells and the production of inflammatory factors in OVA-treated mice. Further investigations demonstrated that CSR treatment relieved AHR in OVA-stimulated mice. The oxidative stress was strengthened in OVA-treated mice, but these effects were relieved by CSR treatment. Lastly, it was discovered that CSR treatment retarded nuclear factor kappa B (NF-κB)/hypoxia-inducible factor 1 alpha (HIF-1α) and p38 mitogen-activated protein kinase (MAPK)/signal transducer and activator of transcription 1 (STAT1) pathways in OVA-triggered asthma mice.. Our findings proved that CSR attenuated the progression of OVA-induced asthma in mice through inhibiting NF-κB/HIF-1α and MAPK/STAT1 pathways. This work might highlight the functions of CSR in the treatment of asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Flavones; Lung; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; NF-kappa B; Ovalbumin; Respiratory Hypersensitivity; STAT1 Transcription Factor

Asthma is a chronic inflammatory disease characterized by airway remodeling and lung inflammation. Collagen triple helix repeat containing 1 (CTHRC1), a glycoprotein, is involved in multiple pathological processes, including inflammation and fibrosis. However, the function of CTHRC1 in asthma remains unclear. In the present study, the mouse asthma model was successfully generated by sensitizing and challenging mice with ovalbumin (OVA). CTHRC1 expression at both RNA and protein levels was significantly upregulated in lung tissues of asthmatic mice. Asthmatic mice exhibited significant airway remodeling as evidenced by increased bronchial wall and smooth muscle cell layer thickness, goblet cell hyperplasia and collagen deposition, and epithelial-mesenchymal transition (EMT), but those characteristics were reversed by CTHRC1 silencing. The cell model with transforming growth factor-β1 (TGF-β1) induction in bronchial epithelial cells (BEAS-2B) was conducted to verify the effects of CTHRC1 on EMT, a classic mechanism that mediates airway remodeling. The results showed that TGF-β1 stimulation increased CTHRC1 expression, and CTHRC1 knockdown inhibited TGF-β1-induced EMT. OVA-treated mice also showed increased inflammatory cell infiltration and the production of OVA-specific immunoglobulin E (IgE), interleukin (IL)-4, IL-5, and IL-13, which were decreased by CTHRC1 downregulation. The effects of CTHRC1 on OVA-induced airway inflammation were further determined by treating BEAS-2B cells with IL-13, in which CTHRC1 knockdown reduced the IL-13-induced secretion of pro-inflammatory factors, including IL-4 and IL-5. In conclusion, these results indicate that CTHRC1 silencing attenuates asthmatic airway remodeling and inflammation in vivo and in vitro, suggesting that CTHRC1 may be a potential target for asthma treatment.

Topics: Airway Remodeling; Animals; Asthma; Collagen; Disease Models, Animal; Inflammation; Interleukin-13; Interleukin-5; Mice; Mice, Inbred BALB C; Ovalbumin; Transforming Growth Factor beta1

Cigarette smoking constitutes a risk factor for severe asthma, which is frequently linked to remodeling of the airways. Appropriate drug treatment for smokers with asthma is uncertain because many smokers with asthma are less sensitive to glucocorticoid treatment than non-smokers with asthma. The purpose of this study was to compare the anti-airway remodeling effects of dexamethasone (Dex) and roflumilast (Rof), a selective phosphodiesterases-4 inhibitor, in smoking and non-smoking mice with asthma. BALB/c mice were sensitized with ovalbumin (OVA) and then challenged with OVA for two weeks, either with or without concurrent exposure to cigarette smoke (CS). Dex (1 mg/kg body weight), Rof (5 mg/kg body weight), or vehicle alone was given orally to the mice once daily. To assess the histopathological effects of airway remodeling, lung tissue sections were obtained. Repeated OVA challenges resulted in fibrosis, goblet cell hyperplasia, and thickening of the airway but not the smooth muscle layer. The presence of CS did not have an impact on the degree of airway remodeling brought on by repeated OVA challenges. In mice repeatedly exposed to OVA either with or without CS, Dex treatment reduced the remodeling alterations. In these mice group, the Rof Treatment had a less significant impact than the Dex treatment. Dex was still more effective than Rof at reducing airway remodeling in asthmatic smoking mice. According to the current study's findings, Dex effectively prevented airway remodeling in a two-week asthma model in mice exposed to CS or not. In contrast, we found that Rof had little to no inhibitory effect of Rof on the airway in our mouse model of asthma, whether or not it had been exposed to CS. We were unable to find solid proof to support CS-induced steroid resistance to treat airway remodeling.

Topics: Animals; Asthma; Body Weight; Cigarette Smoking; Dexamethasone; Disease Models, Animal; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Allergic asthma is a chronic inflammatory illness of the respiratory system characterized by an increase in the number of inflammatory cells in the airways and trouble breathing. Mesenchymal stem cells (MSCs) have the potential to be used in inflammatory diseases as a cellular immunosuppressive treatment. They express calcitriol receptors and communicate with other immunocytes, which increases their anti-inflammatory activity. This study aimed to determine the effects of calcitriol-treated MSC treatment on allergic asthma pathways in a mouse model.. To generate a mouse model of asthma, the mice were sensitized intraperitoneally with ovalbumin (OVA) and aluminum hydroxide emulsion and then challenged intra-nasally with OVA. On day 14, experimental mice received tail vein injections of calcitriol-treated MSCs in PBS prior to allergen exposure. The cytokines assays including IL-4, 10, 12, 17, TGF-β and IFN-γ, splenocytes proliferation, and histological examination of lungs samples were performed. The mice were sensitized with OVA and the response to dexamethasone treatment was compared.. Calcitriol-treated MSCs significantly increased the levels of IL-12, TGF-β, and IFN-γ compared to non-treated MSCs groups. Moreover, calcitriol-treated and non-treated MSCs significantly decreased IL-4 and IL-17 compared to asthmatic groups. The results of the histopathological examination showed that calcitriol-treated MSCs reduced the accumulation of inflammatory cells and bronchial wall thickening in comparison with the asthma group.. Using the allergic asthma model, we were able to show that calcitriol-treated MSCs had an inhibitory impact on airway inflammation. Our findings suggest that the injection of calcitrioltreated MSCs may be a viable treatment option for allergic asthma.

Topics: Animals; Asthma; Calcitriol; Cytokines; Disease Models, Animal; Immunomodulation; Interleukin-4; Lung; Mesenchymal Stem Cells; Mice; Ovalbumin; Transforming Growth Factor beta

Allergic asthma is an inflammatory and chronic condition, which is the most common asthma phenotype. It is usually defined by sensitivity to environmental allergens and leads to the narrowing of the airways. Around 300 million individuals are suffering from asthma worldwide. The purpose of the current research was to evaluate the effect of perillyl alcohol (PA) on oxidative stress and inflammation parameters in rats with allergic asthma. Experimental asthma was induced by ovalbumin (OVA) sensitization and inhalation in five groups of rats including control, asthma, asthma + vehicle, asthma + PA, and asthma + dexamethasone (Dexa). PA (50 mg/kg) or Dexa (2.5 mg/kg) were administered intraperitoneally for seven consecutive days following asthma induction. Histopathological evaluation was performed via hematoxylin and eosin (H&E) and Masson's trichrome staining. The enzyme-linked immunosorbent assay (ELISA) was used for the evaluation of the cytokine levels, including tumor necrosis factor alpha (TNF-α), interleukin (IL)-6, IL-17, and IL-10, as well as oxidative stress biomarkers including reactive oxygen species (ROS), superoxide dismutase (SOD), malondialdehyde (MDA), total antioxidant capacity (TAC), and glutathione peroxidase (GPx) in the lung tissue and bronchoalveolar lavage fluid (BALF). Real-time polymerase chain reaction (PCR) was utilized for assessing the mRNA expression of FOXP3 and GATA3 and western blot analysis was used for the measurement of nuclear factor kappa B (NF-κB) protein expression. PA and Dexa decreased the pathological alterations and the expression levels of inflammatory factors (cytokines, GATA3, and NF-κB) in the lung tissue and BALF of asthmatic rats. PA restored GPx, SOD, and TAC levels and reduced ROS, MDA, nitrite, and total protein in the lung and BALF. Overall, our findings demonstrated that PA can be used as a therapeutic agent in asthma patients, but it is essential to monitor its effects in future clinical studies.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Oxidative Stress; Rats; Reactive Oxygen Species; Superoxide Dismutase

Chronic airway diseases such as asthma are characterized by persistent type 2 immunity in the airways. We know little about the mechanisms that explain why type 2 inflammation continues in these diseases.. We used mouse models to investigate the mechanisms involved in long-lasting immune memory.. Naive mice were exposed intranasally to ovalbumin (OVA) antigen with Alternaria extract as an adjuvant. Type 2 memory was analyzed by parabiosis model, flow cytometry with in vivo antibody labeling, and intranasal OVA recall challenge. Gene-deficient mice were used to analyze the mechanisms.. In the parabiosis model, mice previously exposed intranasally to OVA with Alternaria showed more robust antigen-specific immune responses and airway inflammation than mice with circulating OVA-specific T cells. After a single airway exposure to OVA with Alternaria, CD69

Topics: Allergens; Animals; Asthma; Cytokines; Disease Models, Animal; Inflammation; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Asthma is often exacerbated by airway infection, and some patients with severe asthma may be unresponsive to conventional corticosteroid treatment. Src family kinases (SFKs) were recently implicated in the inflammatory responses of mice induced by allergen and bacterial toxin lipopolysaccharide (LPS). Therefore, we examined the effects of dasatinib (DAS), a Src inhibitor, on airway inflammation in mice induced by ovalbumin (OVA) and LPS. Male A/J mice were sensitized to OVA Day -14 and -7, challenged with intranasal OVA on Day 0, 2, 4, 6 and 8, and on Day 10, mice were also challenged with OVA via inhalation. Mice were treated intranasally with DAS or fluticasone propionate (FP), a glucocorticoid, twice daily for 3 d starting 1 d after OVA inhalation. Moreover, some mice were also administrated LPS 2 h after DAS or FP treatment to model of asthma exacerbation. One day after the last intervention, lung tissue and bronchoalveolar lavage fluid (BALF) were collected. DAS attenuated the accumulation of inflammatory cells and cytokines/chemokines in BALF induced by both OVA and OVA+LPS, while FP did not reduce accumulations induced by OVA+LPS. Therefore, targeting SFKs may be a superior therapeutic approach for asthma exacerbation by infection.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Cytokines; Dasatinib; Disease Models, Animal; Fluticasone; Lipopolysaccharides; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin

Asthma is characterized by chronic inflammation and airway remodeling. However, limited study is conducted on the gene expression profiles of ovalbumin (OVA) induced asthma in mice. Here, we explored the gene expression profiles in lung tissues from mice with OVA-induced asthma using microarray and bioinformatics analysis.. For establishment of OVA-induced asthma model, mice first received intraperitoneal sensitization with OVA on day 0, 7 and 14, followed by atomizing inhalation of OVA 3 times a week for 8 weeks. The lung tissues were collected and subjected to microarray analysis, bioinformatics analysis and expression validation.. Microarray data of lung tissues suggested that 3754 lncRNAs and 2976 mRNAs were differentially expressed in lung tissues between control and asthmatic mice, including 1647 up-regulated and 2106 down-regulated lncRNAs, and 1201 up-regulated and 1766 down-regulated mRNAs. GO analysis displayed that the up-regulated genes were enriched in inflammatory response, leukocyte migration involved in inflammatory response, and Notch signaling pathway. KEGG pathway analysis indicated that the enriched pathway terms of the up-regulated gene included Toll-like receptor signaling pathway and Th17 cell differentiation signaling pathway. Additionally, based on the previously published literatures on asthma and inflammation, we screened out down-regulated genes, such as Smg7, Sumo2, and Stat5a, and up-regulated genes, such as Myl9, Fos and Tlr4. According to the mRNA-lncRNA co-expression network, we selected lncRNAs associated with above genes, including the down-regulated lncRNAs of NONMMUT032848, NONMMUT008873, NONMMUT009478, and NONMMUT006807, and the up-regulated lncRNAs of NONMMUT052633, NONMMUT05340 and NONMMUT042325. The expression changes of the above genes were validated in lung tissues by real-time quantitaive PCR and Western blot.. Overall, we performed gene microarray on lung samples from OVA-induced asthmatic mice and summarized core mRNAs and their related lncRNAs. This study may provide evidence for further research on the therapeutic targets of asthma.

Topics: Animals; Asthma; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Long Noncoding; Transcriptome

Apolipoprotein E (ApoE) is a corticosteroid-unresponsive gene that negatively regulates ovalbumin (OVA) -induced allergic airway inflammation in mice with acute asthma. However, whether ApoE negatively regulates airway remodeling in mice with OVA-induced chronic asthma remains unknown. This study aimed to investigate the effects of ApoE on OVA-induced chronic asthma in a murine model. ApoE knockout (ApoE

Topics: Airway Remodeling; Animals; Apolipoproteins; Apolipoproteins E; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Aluminum oxide nanoparticles (Al

Topics: Aluminum Oxide; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Humans; Lung; Metal Nanoparticles; Mice; Mice, Inbred BALB C; Myeloid Differentiation Factor 88; NF-kappa B; NF-KappaB Inhibitor alpha; Ovalbumin; Phosphorylation; Toll-Like Receptor 4

Asthma poses a significant threat to public health, with an estimated burden of over 334 million people worldwide. Available treatments are often inadequate. We developed a thermo-sensitive hydrogel vaccine containing allergen and FK506 that induced immune tolerance via intranasal administration to treat experimental allergic asthma. The hydrogel delivery system was formulated based on Poloxamer 407 (P407), Carbopol 974P NF, and Polyoxyl 15 hydroxystearate (Kolliphor HS15, HS15). It flowed freely at room temperature and rapidly formed a hydrogel in the nasal cavity once the temperature rose over 33 °C. Ovalbumin and FK506 were slowly released from the hydrogel form and their mucosal residence time was significantly prolonged compared to the liquid formulation. In both an OVA-induced asthma model and an HDM-induced asthma model, the vaccines formulated in hydrogel gave lower levels of eosinophilic inflammation, and airway remodeling. The reduction of lung function was ameliorated, and Foxp3-expressing CD4 + Treg cells were significantly higher. The frequency of Foxp3 + Tregs in lung-draining lymph nodes (dLNs) was correlated with the amelioration. Depletion of Foxp3 + Treg cells abolished the beneficial effects of the allergen/FK506 hydrogel vaccinations. Thus, the allergen/FK506 hydrogel formulation has the potential to be a delivery system for therapeutic allergy vaccines to induce immune tolerance.

Topics: Allergens; Asthma; Disease Models, Animal; Forkhead Transcription Factors; Humans; Hydrogels; Immunotherapy; Ovalbumin; T-Lymphocytes, Regulatory; Tacrolimus; Vaccines

Patients with severe asthma respond poorly to corticosteroids, and their care accounts for more than 60% of the total costs attributed to asthma. Neutrophils form neutrophil extracellular traps (NETs), which play a crucial role in severe asthma. Statins have shown anti-inflammatory effects by reducing NETosis. In this study, we investigate if simvastatin can attenuate severe asthma by reducing NETosis and the underlying mechanism.. Mice were concomitantly sensitized with ovalbumin (OVA), house dust mite (HDM), and lipopolysaccharide (LPS) during sensitization to establish a mouse model of severe asthma with neutrophil predominant inflammation (OVA+LPS mice) and treated with or without simvastatin. In inflammatory response, proportions of Th2, Th17, and Treg cells in lung tissue were detected by flow cytometry, and the levels of cytokines, dsDNA, and MPO-DNA in bronchoalveolar lavage fluid (BALF) were analyzed by ELISA. Citrullinated histone H3 (CitH3) and peptidyl arginine deiminase 4 (PAD4) in lung tissue were determined by Western blot and immunofluorescence imaging. PAD4 mRNA was determined by quantitative PCR (qPCR). HL-60 cells were differentiated into neutrophil-like cells by 1.25% DMSO. The neutrophil-like cells were treated with or without LPS, and simvastatin was then stimulated with PMA. CitH3 and PAD4 expressions were determined.. Sensitization with OVA, HDM, and LPS resulted in neutrophilic inflammation and the formation of NETs in the lungs. Simvastatin treatment reduced the inflammation score, cytokine levels, total cells, and neutrophil counts in the BALF and reduced proportions of Th2 and Th17 but increased Treg cells in lungs of OVA+LPS mice. Simvastatin-treated OVA+LPS mice show reduced NET formation in BALF and lung tissue compared to control mice. Adoptive transfer of neutrophils was sufficient to restore NETosis and neutrophilic inflammation in simvastatin-treated OVA+LPS mice. Simvastatin reduced PAD4 mRNA and protein expression in lung tissues and neutrophils isolated from lungs of OVA+LPS mice and consequent NET formation. In vitro, simvastatin reduced LPS-induced PAD4 upregulation and NETosis in HL-60-differentiated neutrophil-like cells. Furthermore, PAD4-overexpressed lentiviral transduction was sufficient to restore PAD4 protein expression and NETosis in simvastatin-treated HL-60-differentiated neutrophil-like cells.. Simvastatin reduces Th17-mediated neutrophilic inflammation and airway hyperreactivity by reducing PAD4 expression and inhibiting NETosis in a mouse model of severe asthma. Severe asthmatic patients with high levels of circulating NETs or sputum NETs may show improved responses to statin treatment.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; DNA; Extracellular Traps; Histones; Inflammation; Lipopolysaccharides; Lung; Mice; Neutrophils; Ovalbumin; RNA, Messenger; Simvastatin

Intrauterine growth restriction (IUGR) is strongly correlated with an increased risk of asthma later in life. Farm dust protects mice from developing house dust mite-induced asthma, and loss of ubiquitin modifying enzyme A20 in lung epithelium would abolish this protective effect. However, the mechanisms of A20 in the development of asthma following IUGR remains unknown.. An IUGR rat model induced by maternal nutrient restriction was used for investigating the role of A20 in the response characteristics of IUGR rats to ovalbumin (OVA) challenge. The ubiquitination of proteins and N6-methyladenosine (m6A) modifications were used to further assess the potential mechanism of A20.. IUGR can reduce the expression of A20 protein in lung tissue of newborn rats and continue until 10 weeks after birth. OVA challenging can increase the expression of A20 protein in lung tissue of IUGR rats, but its level was still significantly lower than the control OVA group. The differentially ubiquitinated proteins in lung tissues were also observed in IUGR and normal newborn rats. Furthermore, this ubiquitination phenomenon continued from the newborn to adulthood. In the detected RNA methylations, m6A abundance of the motif GGACA was the highest. The higher abundances of m6A modification of A20 mRNA from IUGR were negatively correlated with the trend of A20 protein levels.. These findings indicate A20 as a key regulator during the development of asthma following IUGR, providing further insight into the prevention of asthma induced by environmental factors.

Topics: Animals; Asthma; Female; Fetal Growth Retardation; Lung; Ovalbumin; Rats; Ubiquitin

Stigmasterol has significant anti-arthritis and anti-inflammatory effects, but its role in immune and inflammatory diseases is still unclear.. The potential advantages of stigmasterol in asthma were explored in IL-13-induced BEAS-2B cells and asthmatic mice.. The optimal target of stigmasterol was confirmed in asthma. After detecting the cytotoxicity of stigmasterol in BEAS-2B cells, 10 μg/mL and 20 μg/mL stigmasterol were incubated with the BEAS-2B cell model for 48 h, and anti-inflammation and antioxidative stress were verified. Asthmatic mice were induced by OVA and received 100 mg/kg stigmasterol for 7 consecutive days. After 28 days, lung tissues and BAL fluid were collected for the following study. To further verify the role of NK1-R, 0.1 μM WIN62577 (NK1-R specific antagonist), and 1 μM recombinant human NK1-R protein were applied.. NK1-R was the potential target of stigmasterol. When the concentration of stigmasterol is 20 μg/mL, the survival rate of BEAS-2B cells is about 98.4%, which is non-toxic. Stigmasterol exerted anti-inflammation and antioxidant stress in a dose-dependent manner and decreased NK1-R expression in IL-13-induced BEAS-2B. Meanwhile,. The protective effect of stigmaterol on asthma and its underlying mechanism have been discussed in depth, providing a theoretical basis and more possibilities for its treatment of asthma.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Humans; Inflammation; Interleukin-13; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Neurokinin-1; Respiratory Hypersensitivity; Stigmasterol

Asthma is a chronic respiratory disease with an increasing incidence every year. microRNAs (miRNAs) have been demonstrated to have implications for asthma. However, limited information is available regarding the effect of miR-124-3p on this disease. Therefore, this study aimed to explore the possible effects of miR-124-3p and S100A4 on inflammation and epithelial-mesenchymal transition (EMT) in asthma using mouse models.. Ovalbumin was used to induce asthmatic mouse models. Lung injury in mouse models was assessed, and the bronchoalveolar lavage fluid of mice was collected to determine the number of eosinophilic granulocytes and assess inflammation. The expression levels of miR-124-3p, S100A4, E-cadherin, N-cadherin, Snail1, vimentin, and TGF-β1/Smad2 signaling pathway-related proteins were measured using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. In vitro experiments, cells were transfected with miR-124-3p mimics or inhibitors to test the expression of S100A4 by RT-qPCR and western blot analysis, and the mutual binding of miR-124-3p and S100A4 was validated by dual-luciferase reporter gene assay.. Overexpression of miR-124-3p or inhibition of S100A4 expression attenuated bronchial mucus secretion and collagenous fibers and suppressed inflammatory cell infiltration. Additionally, upon miR-124-3p overexpression or S100A4 suppression, eosinophilic granulocytes were decreased, interleukin-4 (IL-4) and IL-13 expression levels were reduced in the bronchoalveolar lavage fluid, serum total IgE level was reduced, and the TGF-β1/Smad2 signaling pathway was suppressed. Mechanically, a dual-luciferase reporter gene assay verified the binding relationship between miR-124-3p and S100A4.. miR-124-3p can negatively target S100A4 to attenuate inflammation in asthmatic mouse models by suppressing the EMT process and the TGF-β/smad2 signaling pathway.

Topics: Animals; Asthma; Inflammation; Mice; MicroRNAs; Ovalbumin; S100 Calcium-Binding Protein A4; Transforming Growth Factor beta1

Obesity increases the severity of airway hyperresponsiveness (AHR) in individuals with asthma, but the mechanism is not well elucidated. G-protein coupled receptor 40 (GPR40) has been found to induce airway smooth muscle contraction after activated by long-chain fatty acids (LC-FFAs), suggesting a close correlation between GPR40 and AHR in obese. In this study, C57BL/6 mice were fed a high-fat diet (HFD) to induce obesity with or without ovalbumin (OVA) sensitization, the regulatory effects of GPR40 on AHR, inflammatory cells infiltration, and the expression of Th1/Th2 cytokines were evaluated by using a small-molecule antagonist of GPR40, DC260126. We found that the free fatty acids (FFAs) level and GPR40 expression were greatly elevated in the pulmonary tissues of obese asthmatic mice. DC260126 greatly reduced methacholine-induced AHR, ameliorated pulmonary pathological changes and decreased inflammatory cell infiltration in the airways in obese asthma. In addition, DC260126 could down-regulate the levels of Th2 cytokines (IL-4, IL-5, and IL-13) and pro-inflammatory cytokines (IL-1β, TNF-α), but elevated Th1 cytokine (IFN-γ) expression. In vitro, DC260126 could remarkedly reduce oleic acid (OA)-induced cell proliferation and migration in HASM cells. Mechanistically, the effects that DC260126 alleviated obese asthma was correlated with the down-regulation of GTP-RhoA and Rho-associated coiled-coil-forming protein kinase 1 (ROCK1). Herein, we proved that targeting of GPR40 with its antagonist helped to mitigate multiple parameters of obese asthma effectively.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Obese; Obesity; Ovalbumin; Receptors, G-Protein-Coupled; Respiratory Hypersensitivity; Signal Transduction

Shaoyao-Gancao Tang (SGT) is a traditional Chinese medicine formulation. It has been used to treat kinds of pain and to alleviate asthma in clinic. However, the mechanism of action is not known.. To investigate the anti-asthma effect of SGT involving modulation of the T-helper type 1 (Th1) Th1/Th2 ratio in the gut-lung axis and alteration of the gut microbiota (GM) in rats with ovalbumin (OVA)-induced asthma.. The main constituents of SGT were analyzed by high-performance liquid chromatography (HPLC). A model of asthma was established in rats by OVA-induced allergen challenge. Rats suffering from asthma (RSAs) were treated with SGT (2.5, 5.0 and 10.0 g/kg), dexamethasone (1 mg/kg) or physiologic saline for 4 weeks. The level of immunoglobulin (Ig)E in bronchoalveolar lavage fluid (BALF) and serum was determined by enzyme-linked immunosorbent assay. Histology of lung and colon tissues was investigated using staining (hematoxylin and eosin and periodic acid-Schiff). The Th1/Th2 ratio and levels of cytokines (interferon (IFN)-γ and interleukin (IL)-4) in the lung and colon were detected by immunohistochemistry. The GM in fresh feces was analyzed by 16 S rRNA gene sequencing.. Twelve main constituents (gallic acid, albiflorin, paeoniflorin, liquiritin apioside, liquiritin, benzoic acid, isoliquiritin apioside, isoliquiritin, liquiritigenin, glycyrrhizic acid, isoliquiritigenin and glycyrrhetinic acid) of SGT were simultaneously determined by HPLC. SGT treatment (5.0 and 10.0 g/kg) was found to reduce the IgE level (a vital marker of hyper-responsiveness) in BALF and serum, improve typical morphological changes (inflammatory-cell infiltration and goblet cell metaplasia) in the lung and colon, alleviate airway remodeling (including bronchiostenosis and basement membrane-thickening) in the lung, significantly decrease the IL-4 level and increase the IFN-γ level in the lung and colon, which led to restoration of the IFN-γ/IL-4 ratio. The dysbiosis and dysfunction of GM in RSAs were modulated by SGT. The abundance of bacteria of the genera Ethanoligenens and Harryflintia was increased in RSAs and was decreased upon SGT treatment. The abundance of Family_XIII_AD3011_group was decreased in RSAs and increased upon SGT treatment. Moreover, SGT therapy increased the abundance of bacteria of the genera Ruminococcaceae_UCG-005 and Candidatus_Sacchrimonas, and decreased that of Ruminococcus_2 and Alistipes.. SGT ameliorated rats with OVA-induced asthma via regulation of the Th1/Th2 ratio in the lung and gut, and modulated the GM.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Gastrointestinal Microbiome; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Rats; Th2 Cells

Corticosteroid resistance, progressive lung function decline, and frequent asthma exacerbations are the hallmarks of neutrophilic asthma (NA). However, the potential contributors and their mechanisms of NA aggravation have not yet been fully clarified. This study was conducted to assess the precise mechanism and inflammatory effects of endocrine-disrupting chemicals using mono-n-butyl phthalate (MnBP) on an NA model. BALB/c mice from normal control and LPS/OVA-induced NA groups were treated with or without MnBP. The effects of MnBP on the airway epithelial cells (AECs), macrophages (Mφ), and neutrophils were investigated in vitro and in vivo. NA mice exposed to MnBP had significantly increased airway hyperresponsiveness, total and neutrophil cell counts in the bronchoalveolar lavage fluid, and the percentage of M1Mφ in the lung tissues compared to those non-exposed to MnBP. In in vitro study, MnBP induced the human neutrophil activation to release neutrophil DNA extracellular traps, Mφ polarizing toward M1Mφ, and AEC damage. Treatment with hydroxychloroquine (an autophagy inhibitor) reduced the effects of MnBP in vivo and in vitro. The results of our study suggest that MnBP exposure may increase the risk of neutrophilic inflammation in severe asthma and autophagy pathway-targeted therapeutics can help control MnBP-induced harmful effects in asthma.

Topics: Animals; Asthma; Autophagy; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Respiratory Hypersensitivity

Asthma often presents with a daily rhythm; however, the underlying mechanisms remain unclear. Circadian rhythm genes have been proposed to regulate inflammation and mucin expression. Here, ovalbumin (OVA)-induced mice and serum shock human bronchial epidermal cells (16HBE) were used in in vivo and in vitro models, respectively. We constructed a brain and muscle ARNT-like 1 (BMAL1) knockdown 16HBE cell line to analyze the effects of rhythmic fluctuations on mucin expression. Serum immunoglobulin E (IgE) and circadian rhythm genes in asthmatic mice showed rhythmic fluctuation amplitude. Mucin (MUC) 1 and MUC5AC expression was increased in the lung tissue of the asthmatic mice. MUC1 expression was negatively correlated with that of the circadian rhythm genes, particularly BMAL1 (r = -0.546, P = 0.006). There was also a negative correlation between BMAL1 and MUC1 expression (r = -0.507, P = 0.002) in the serum shock 16HBE cells. BMAL1 knockdown negated the rhythmic fluctuation amplitude of MUC1 expression and upregulated MUC1 expression in the 16HBE cells. These results indicate that the key circadian rhythm gene, BMAL1, causes periodic changes in airway MUC1 expression in OVA-induced asthmatic mice. Targeting BMAL1 to regulate periodic changes in MUC1 expression may, therefore, improve asthma treatments.

Topics: Animals; ARNTL Transcription Factors; Asthma; Brain; Humans; Mice; Mucin-1; Mucins; Muscles; Ovalbumin

Asthma is an important pulmonary disease associated with T helper lymphocyte (Th)2 dominant immune response, which can initiate allergic and inflammatory reactions. Interleukin (IL)-10 is the main immune suppressor cytokine, and mesenchymal stem cells (MSCs) have an immune-modulatory potential that can be transduced with the expression of the IL-10 gene to control pathophysiology of allergic asthma. Bone marrow's MSCs were isolated and transduced with the expression vector that contains the expressible IL-10 gene. Then, allergic asthma mouse model was produced and treated with manipulated MSCs. Methacholine challenge test; measurement of IL-4, IL-5, IL-8, IL-13, IL-25, and IL-33; and total and ovalbumin (OVA)-specific immunoglobulin (Ig)E levels were done. Hyperplasia of the goblet cell, secretion of mucus, and peribronchiolar and perivascular eosinophilic inflammation were evaluated in lung pathological sections. IL-25, IL-33, and total IgE levels; AHR; eosinophilic inflammation; hyperplasia of the goblet cell; and secretion of mucus could be controlled in M, MV, and MV-10 groups, and the control in the MV-10 group was strong compared to M and MV groups. MSCs have immune-modulatory capacity that can control allergic asthma pathophysiology, and this effect can be strengthened and reinforced by the expression of IL-10 gene.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Eosinophilia; Hyperplasia; Immunoglobulin E; Inflammation; Interleukin-10; Interleukin-33; Lung; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Ovalbumin

Topics: Animals; Asthma; Cadmium; Endoribonucleases; Hyperplasia; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Protein Serine-Threonine Kinases

Glycolipids with TLR4 agonistic properties can serve either as therapeutic agents or as vaccine adjuvants by stimulating the development of proinflammatory responses. Translating them to the clinical setting is hampered by synthetic difficulties, the lack of stability in biological media, and/or a suboptimal profile of balanced immune mediator secretion. Here, we show that replacement of the sugar fragment by an sp

Topics: Adjuvants, Immunologic; Adjuvants, Pharmaceutic; Animals; Asthma; CD8-Positive T-Lymphocytes; Cysteine; Cytokines; Disease Models, Animal; Immunotherapy; Mice; Mice, Inbred BALB C; Ovalbumin; Serine; Th2 Cells; Toll-Like Receptor 4

Overproduction of pro-inflammatory cytokines and its-mediated immune cell infiltration play a crucial role in asthma progression. In this study, we investigated the role of ginsenoside Rh1 (Rh1) in ovalbumin (OVA)/lipopolysaccharide (LPS)-induced allergic asthma both in vitro and in vivo.. The phorbol ester (PMA) and LPS were used to induce inflammation in lung airway cells and macrophage activation, respectively. Western blotting, quantitative reverse transcription-PCR, and immunofluorescence (IF) assays were performed to elucidate the underlying molecular mechanisms. To evaluating the effects of Rh1 in vivo, OVA and LPS were used to establish allergic asthma models.. Rh1 significantly suppressed PMA-induced lung inflammation and macrophage activation by suppressing pro-inflammatory cytokines (TNF-α, IL-1β, MCP-1), ICMA-1, and matrix metallopeptidase 9 (MMP9) in A549 cells. Rh1 abolished the PMA-induced inflammation by suppressing MAPK, Akt, and NF-κB p65. Pretreatment with Rh1 blocked PMA-mediated translocation of NF-κB, a key marker of pro-inflammatory cytokine release, into the nucleus. Similar to PMA-induced lung inflammation, Rh1 suppressed LPS-induced macrophage activation by suppressing NF-κB p65 activation and inducible nitric oxide synthase protein and mRNA expression. Consistent with in vitro data, LPS injection enhanced the number of immune cells induced by OVA in bronchoalveolar lavage fluid, whereas 20 mg/kg Rh1 significantly decreased OVA/LPS-mediated immune cell induction. In addition, Rh1 inhibited eosinophil, macrophage, and neutrophil maturation through by IL-4 and OVA-specific IgE production.. Rh1 protects against OVA/LPS-induced allergic asthma by suppressing immune cell infiltration by blocking the activation of MAPK, Akt, and NF-κB signaling pathways.

Topics: Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Humans; Inflammation; Lipopolysaccharides; Lung; NF-kappa B; Ovalbumin; Pneumonia; Proto-Oncogene Proteins c-akt; Signal Transduction

To investigate the role of the costimulatory molecule CD226 in asthma pathogenesis, we produced a CD4

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Cytokines; Disease Models, Animal; Gastrointestinal Microbiome; Interleukin-10; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin

Dibutyl phthalate (DBP), used as a plasticizer, is of wide concern as an environmental pollutant since it has certain immunotoxicity. Although there is growing evidence supporting a link between DBP exposure and allergic airway inflammation, there is less information concerned with whether the ferroptosis pathway is involved in DBP-aggravated allergic asthma in ovalbumin (OVA)-sensitized mice. This study aimed to investigate the role and underlying mechanisms of ferroptosis in DBP-exposed allergic asthmatic mice. Balb/c mice were orally exposed to 40 mg/kg

Topics: Animals; Asthma; Dibutyl Phthalate; Ferroptosis; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Reactive Oxygen Species

Polysaccharides, as one of the main types of bioactive components of Cordyceps militaris, have anti-allergic asthma effects. Herein, an ovalbumin-induced allergic asthma mouse model was established to assess the potential mechanisms of the separated and purified Cordyceps militaris polysaccharide (CMP). CMP is an α-pyranose with a molecular weight of 15.94 kDa that consists of Glc, Man, Gal, Xyl, Ara and GlcA in a molar ratio of 81.25:21.96:13.88:3.92:3.58:1.00. CMP improved inflammatory cytokine levels, alleviated the histopathological changes in the lung and intestinal tissues, regulated the expression of mRNA and proteins related to oxidative stress and inflammatory pathways, reversed gut dysbiosis at the phylum and family levels and improved microbiota function in allergic asthma mice. Moreover, it was found that the levels of inflammatory cytokines in lung tissue of mice were significantly correlated with some intestinal microbial communities. Overall, CMP improved oxidative stress and the inflammatory response in allergic asthma mice by regulating the Nrf2/HO-1 and NF-κB signaling pathways, which may be closely correlation with maintaining the stability of the gut microbiota.

Topics: Animals; Asthma; Cordyceps; Cytokines; Gastrointestinal Microbiome; Mice; NF-E2-Related Factor 2; NF-kappa B; Ovalbumin; Polysaccharides; Signal Transduction

Topics: Animals; Anti-Inflammatory Agents; Asthma; Autophagy; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity

Topics: Alveolitis, Extrinsic Allergic; Animals; Asthma; Dexamethasone; Inflammation; Macrophages; Male; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Phenotype; Pneumonia; Rats; Rats, Wistar

Sensitive skin, a common pathophysiological feature of allergic diseases, is defined as an unpleasant sensation in response to stimuli that normally should not provoke such sensations. However, the relationship between allergic inflammation and hypersensitive skin in the trigeminal system remains to be elucidated. To explore whether bronchial allergic inflammation affects facial skin and primary sensory neurons, we used an ovalbumin (OVA)-induced asthma mouse model. Significant mechanical hypersensitivity was observed in the facial skin of mice with pulmonary inflammation induced by OVA sensitization compared to mice treated with adjuvant or vehicle as controls. The skin of OVA-treated mice showed an increased number of nerve fibers, especially rich intraepithelial nerves, compared to controls. Transient receptor potential channel vanilloid 1 (TRPV1)-immunoreactive nerves were enriched in the skin of OVA-treated mice. Moreover, epithelial TRPV1 expression was higher in OVA-treated mice than in controls. Trigeminal ganglia of OVA-treated mice displayed larger numbers of activated microglia/macrophages and satellite glia. In addition, more TRPV1 immunoreactive neurons were found in the trigeminal ganglia of OVA-treated mice than in controls. Mechanical hypersensitivity was suppressed in OVA-treated Trpv1-deficient mice, while topical skin application of a TRPV1 antagonist before behavioral testing reduced the reaction induced by mechanical stimulation. Our findings reveal that mice with allergic inflammation of the bronchi had mechanical hypersensitivity in the facial skin that may have resulted from TRPV1-mediated neuronal plasticity and glial activation in the trigeminal ganglion.

Topics: Animals; Antineoplastic Agents; Asthma; Inflammation; Mice; Ovalbumin; Skin; TRPV Cation Channels

Topics: Airway Remodeling; Animals; Asthma; Cytokines; Disease Models, Animal; Inflammation; Interleukin-33; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin

This study aimed to investigate the effects of sulfur dioxide (SO2) derivatives on asthma induced by ovalbumin (OVA). Sprague Dawley rats were sensitized to and challenged with OVA and SO2 derivatives (NaHSO3 and Na2SO3, 1:3 M/M) to establish 28-day (short-term) and 42-day (long-term) asthma models. Exposure to SO2 derivatives aggravated asthma and hence, promoted lung injury in OVA-induced asthma. In addition, it upregulated the protein expression of TRPV1 and downregulated the expression of tight junctions (TJs). These changes were dose-dependent and were more pronounced in the presence of a high concentration of SO2 derivatives. In vitro, SO2 derivatives also increased the calcium influx and TRPV1 protein expression, and decreased TJ expression. Besides, no significant difference in the TJ expression was found between the WT and TRPV1-/- mice. The underlying mechanism might be related to regulating the effects of TRPV1 and TJs.

Topics: Animals; Asthma; Disease Models, Animal; Lung; Mice; Ovalbumin; Rats; Rats, Sprague-Dawley; Sulfur Dioxide; Tight Junctions; TRPV Cation Channels

Asthma is a chronic airway inflammatory disease. Current treatment of mainstream medications has significant side effects. There is growing evidence that the refractoriness of asthma is closely related to common changes in the lung and intestine. The lungs and intestines, as sites of frequent gas exchange in the body, are widely populated with gas signaling molecules NO and CO, which constitute NO-CO metabolism and may be relevant to the pathogenesis of asthma in the lung and intestine. The Chinese herbal formula Tingli Dazao Xiefei Decoction (TD) is commonly used in clinical practice to treat asthma with good efficacy, but there are few systematic evaluations of the efficacy of asthma on NO-CO metabolism, and the mode of action of its improving effect on the lung and intestine is unclear.. To investigate the effect of TD on the lung and intestine of asthmatic rats based on NO-CO metabolism.. In vivo, we established a rat asthma model by intraperitoneal injection of sensitizing solution with OVA atomization, followed by intervention by gavage administration of TD. We simultaneously examined alterations in basal function, pathology, NO-CO metabolism, inflammation and immune cell homeostasis in the lungs and intestines of asthmatic rats, and detected changes in intestinal flora by macrogenome sequencing technology, with a view to multi-angle evaluation of the treatment effects of TD on asthmatic rats. In vitro, lung cells BEAS-2B and intestinal cells NCM-460 were used to establish a model of lung injury causing intestinal injury using LPS and co-culture chambers, and lung cells or intestinal cells TD-containing serum was administered to intervene. Changes in inflammatory, NO-CO metabolism-related, cell barrier-related and oxidative stress indicators were measured in lung cells and intestinal cells to evaluate TD on intestinal injury by way of amelioration and in-depth mechanism.. In vivo, our results showed significant basal functional impairment in the lung and intestine of asthmatic rats, and an inflammatory response, immune cell imbalance and intestinal flora disturbance elicited by NO-CO metabolic disorders were observed (P < 0.05 or 0.01). The administration of TD was shown to deliver a multidimensional amelioration of the impairment induced by NO-CO metabolic disorders (P < 0.05 or 0.01). In vitro, the results showed that LPS-induced lung cells BEAS-2B injury could cause NO-CO metabolic disorder-induced inflammatory response, cell permeability damage and oxidative stress damage in intestinal cells NCM-460 (P < 0.01). The ameliorative effect on intestinal cells NCM-460 could only be exerted when TD-containing serum interfered with lung cells BEAS-2B (P < 0.01), suggesting that the intestinal ameliorative effect of TD may be exerted indirectly through the lung.. TD can ameliorate NO-CO metabolism in the lung and thus achieve the indirectly amelioration of NO-CO metabolism in the intestine, ultimately achieving co-regulation of lung and intestinal inflammation, immune imbalance, cellular barrier damage, oxidative stress and intestinal bacterial disorders in asthma in vivo and in vitro. Targeting lung and intestinal NO-CO metabolic disorders in asthma may be a new therapeutic idea and strategy for asthma.

Topics: Animals; Asthma; Disease Models, Animal; Inflammation; Intestinal Diseases; Intestines; Lipopolysaccharides; Lung; Metabolic Diseases; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Rats

Arisaema cum Bile (Dan Nanxing in Chinese, DNX) have been employed to treat allergic asthma. However, the active components and its mechanisms remain unknown. Therefore, the systematic pharmacology approach-experimental validation was performed in this study. Each 5, 6, and 10 compounds of DNX were obtained by HPLC analysis, TCMSP, and literature report, respectively. A total of 379 targets on all these compounds were acquired from Swiss Target Prediction, and 1973 targets on allergic asthma were predicated. The KEGG enrichment analysis was performed. Furthermore, a rat model of allergic asthma was established and DNX (450 mg/kg, p.o.) was given for 2 weeks. DNX treatment prevented OVA-induced pathological changes in lung cell of irregular arrange and necrotic bronchial epithelial. It also decreased inflammatory cytokines IL-4, IL-5, and IL-13 of serum and BALF, and increased IL-12 and IFN-γ. The main MAPK signaling pathway predicted by KEGG enrichment was verified, as indicated by the decreased protein expression of JNK (p < 0.05 & p < 0.01), ERK (p < 0.05), and p38 MAPK (p < 0.01) in lung tissue. These findings indicated that DNX attenuated OVA-induced allergic asthma mainly by decreasing the MAPK signaling pathway.

Topics: Animals; Arisaema; Asthma; Bile; Cytokines; Disease Models, Animal; Mice; Mice, Inbred BALB C; Molecular Structure; Network Pharmacology; Ovalbumin; Rats

Asthma is a heterogeneous, chronic respiratory disease characterized by airway inflammation and remodeling. Phosphodiesterase (PDE) inhibitors represent one of the intensively studied groups of potential anti-asthmatic agents due to their affecting both airway inflammation and remodeling. However, the effect of inhaled pan-PDE inhibitors on allergen induced asthma has not been reported to date. In this study we investigated the impact of two, representative strong pan-PDE inhibitors from the group of 7,8-disubstituted derivatives of 1,3-dimethyl-3,7-dihydro-1H-purine-2,6-dione: compound 38 and 145, on airway inflammation and remodeling in murine model of ovalbumin (OVA)-challenged allergic asthma. Female Balb/c mice were sensitized and challenged with OVA, 38 and 145 were administrated by inhalation, before each OVA challenge. The inhaled pan-PDE inhibitors markedly reduced the OVA-induced airway inflammatory cell infiltration, eosinophil recruitment, Th2 cytokine level in bronchoalveolar lavage fluid, as well as both, total and OVA-specific IgE levels in plasma. In addition, inhaled 38 and 145 decreased many typical features of airway remodeling, including goblet cell metaplasia, mucus hypersecretion, collagen overproduction and deposition, as well as Tgfb1, VEGF, and α-SMA expression in airways of allergen challenged mice. We also demonstrated that both 38 and 145 alleviate airway inflammation and remodelling by inhibition of the TGF-β/Smad signaling pathway activated in OVA-challenged mice. Taken together, these results suggest that the investigated pan-PDE inhibitors administered by inhalation are dual acting agents targeting both airway inflammation and remodeling in OVA-challenged allergic asthma and may represent promising, anti-asthmatic drug candidates.

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphodiesterase Inhibitors

One of the inflammatory diseases of the respiratory system is asthma. Teucrium polium (TP) has anti-inflammatory and anti-allergic properties and its anti-asthmatic effects have not been investigated yet. RORγt is an inflammatory transcription factor for Th17 differentiation. By secreting IL-17, Th17 leads to neutrophilic inflammation in the lungs. As an anti-inflammatory cytokine, IL-10 reduces the dissemination of inflammatory elements in the airways.. To evaluate the effect of TP extract in asthma treatment.. Thirty female Balb/c mice were distributed into 5 groups (n=6) including the control, treated with ovalbumin (OVA), and OVA+ various doses of TP (50, 150, and 300 mg/kg). All groups except the control group were sensitized to OVA solution on days 0, 7, and 14 by subcutaneous injection. The challenge was performed on days 18 to 21 by the inhalation of 1% OVA and the treatment was done with TP extract in the treatment groups, half an hour before the challenge. On day 22, the serum and spleen samples were collected to determine IL-10 serum levels and RORγt gene expression, respectively.. In the treatment groups, the expression of RORγt significantly decreased when using OVA+ Tp extract (150 mg/kg and 300 mg/kg), and IL-10 serum levels significantly increased when using OVA+ TP extract (150 mg/kg) compared with the OVA group.. It is possible that TP extract can be effective in improving asthma by reducing inflammation.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Inflammation; Interleukin-10; Lung; Mice; Mice, Inbred BALB C; Nuclear Receptor Subfamily 1, Group F, Member 3; Ovalbumin; Teucrium

Particulate matter (PM) is a major component of air pollution from emissions from anthropogenic and natural sources and is a serious problem worldwide due to its adverse effects on human health. Increased particulate air pollution increases respiratory disease-related mortality and morbidity. However, the impact of PM with an aerodynamic diameter of ≤ 2.5 μm (PM2.5) on combined allergic rhinitis and asthma syndrome (CARAS) remains to be elucidated. Accordingly, in the present study, we investigated the effect of PM2.5 in an ovalbumin (OVA)-induced CARAS mouse model with a focus on NF-κB signaling.. We established an OVA-induced mouse model of CARAS to determine the effects of exposure to PM2.5. BALB/c mice were randomly divided into four groups: (1) naive, (2) PM2.5, (3) CARAS, and (4) CARAS/PM2.5. Mice were systemically sensitized with OVA and challenged with inhalation of ultrasonically nebulized 5% OVA three times by intranasal instillation of OVA in each nostril for 7 consecutive days. Mice in the PM2.5 and CARAS/PM2.5 groups were then exposed to PM2.5 by intranasal instillation of PM2.5 for several days. We then examined the impacts of PM2.5 exposure on histopathology and NF-κB signaling in our OVA-induced CARAS mouse model.. Our results demonstrate that PM2.5 can aggravate OVA-induced CARAS.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Humans; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Particulate Matter; Rhinitis, Allergic; Signal Transduction

To investigate the role of neferine in ovalbumin (OVA)-induced asthma, and to reveal the possible mechanism.. Neferine relieves asthma-induced inflammatory reaction, airway resistance, and lung injury by inhibiting MAPK signaling pathways. This could serve neferine as a novel therapeutic candidate for treating asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Immunoglobulin E; Inflammation; Interleukin-13; Interleukin-5; Interleukin-6; Lung; Lung Injury; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Ovalbumin; Tumor Necrosis Factor-alpha

Asthma is a common chronic inflammatory disease of the airways with no known cure. Lipid mediators (LMs) are a kind of inflammatory signaling molecules which are believed to be involved in the development of asthma.. Anti-inflammatory effect of SXCF and RosA was assessed using OVA-induced asthma model mice by UPLC-MS/MS method.. Overall, RosA played a critical role in anti-asthma treatment. In total, 90% of LMs species that were significantly regulated by SXCF were covered. On the most important LMs associated with asthma, RosA equivalent induced similar effects as SXCF did. It is believed that some constituents in SXCF could neutralize RosA excessive impacts on LMs.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Chromatography, Liquid; Cytokines; Disease Models, Animal; Hyssopus Plant; Lipids; Mice; Mice, Inbred BALB C; Ovalbumin; Rosmarinic Acid; Tandem Mass Spectrometry

Allergies are increasing worldwide. The presence of atopic diseases in the mother propagates the onset of allergic diseases in the offspring with a considerably stronger penetrance than atopic diseases of the father. Such observation challenges genetic predispositions as the sole cause of allergic diseases. Epidemiological studies suggest that caregiver stress in the perinatal period may predispose offspring to asthma. Only one group has studied the link between prenatal stress and neonatal asthma susceptibility in a murine model.. We aimed to study if the neonatal increased risk of developing allergic lung inflammation persists after puberty and if there are sex differences in susceptibility.. Pregnant BALB/c mice were subjected to a single restraint stress exposure at day 15 of gestation. Pups were separated by gender and subjected to a well-known sub-optimal asthma model after puberty.. Adult mice born to stressed dams were more susceptible to developing allergic pulmonary inflammation since an increase in the number of eosinophils in bronchoalveolar lavage (BAL), a greater peribronchial and perivascular infiltrate, a higher proportion of mucus-producing cells, and increased IL-4 and IL-5 levels in BAL were detected compared to control mice. These effects were more profound in females than males. Moreover, only females from stressed dams showed an increase in IgE levels.. Increased litter susceptibility to develop allergic lung inflammation induced by maternal stress persists after puberty and is more potent in females than in male mice.

Topics: Animals; Asthma; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Pregnancy

Asthma is an inflammatory reaction mediated by type 2 helper T (Th2) cells and is known to increase eosinophil levels. Our previous study showed that stress-related asthma can cause neutrophilic and eosinophilic airway inflammation by suppressing immune tolerance. However, the mechanism of stress-induced neutrophilic and eosinophilic airway inflammation remains unclear. Therefore, to elucidate the cause of neutrophilic and eosinophilic inflammation, we investigated the immune response during the induction of airway inflammation. In addition, we focused on the relationship between immune response modulation immediately after stress exposure and the development of airway inflammation.. Asthmatic mice were induced by three phases using female BALB/c mice. During the first phase, the mice were made to inhale ovalbumin (OVA) to induce immune tolerance before sensitization. Some mice were exposed to restraint stress during the induction of immune tolerance. In the second phase, the mice were sensitized with OVA/alum intraperitoneal injections. In the final phase, onset of asthma was induced through OVA exposure. Asthma development was evaluated based on airway inflammation and T-cell differentiation. Microarray and qPCR analyses were used to enumerate candidate factors to investigate the starting point of immunological modification immediately after stress exposure. Furthermore, we focused on interleukin-1β (IL-1β), which initiates these immune modifications, and performed experiments using its receptor blocker interleukin-1 receptor antagonist (IL-1RA).. Stress exposure during immune tolerance induction increased eosinophil and neutrophil airway infiltration. This inflammation was associated with decreased T regulatory cell levels and increased Th2 and Th17 levels in bronchial lymph node cells. Microarray and qPCR analyses showed that the initiation of Th17 differentiation might be triggered by stress exposure during tolerance induction. IL-1RA administration during stress exposure suppressed neutrophilic and eosinophilic airway inflammation via Th17 reduction and Treg increase.. Our results show that psychological stress causes both eosinophilic and neutrophilic inflammatory responses due to the breakdown of immune tolerance. Furthermore, stress-induced inflammation can be abolished using IL-1RA.

Topics: Animals; Asthma; Disease Models, Animal; Female; Immune Tolerance; Immunity; Inflammation; Interleukin 1 Receptor Antagonist Protein; Interleukin-1beta; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Stress, Psychological; Th17 Cells; Th2 Cells

Fixed airflow limitation (FAO), prevalent in patients with severe or difficult-to-treat asthma, is mainly caused by airway remodeling. Airway remodeling is initiated by inflammation and involves subsequent pathological changes. Glycyl-l-histidyl-l-lysine (GHK) is a matrikine with anti-inflammatory and antioxidant effects, naturally existing in human tissue. At present, the GHK level in human plasma and whether it is related to airway remodeling of asthma remain unclear. This study was conducted to determine how GHK is involved in airway remodeling in asthma. Our result showed that the plasma GHK levels of patients with asthma were significantly lower than those of age-matched healthy controls. In asthma patients, plasma GHK levels display a moderate correlation with FEF

Topics: Airway Remodeling; Animals; Asthma; Disease Models, Animal; Epithelial Cells; Humans; Lysine; Mice; Ovalbumin; Sirtuin 1; Transforming Growth Factor beta1

Combined allergic rhinitis and asthma syndrome (CARAS) causes chronic respiratory inflammation in allergic individuals. Long-term exposure to particulate matter 2.5 (PM2.5; particles 2.5 µm or less in diameter) can aggravate respiratory damage. Bergapten (5-methoxysporalen) is a furocoumarin mostly found in bergamot essential oil and has significant antioxidant, anticancer, and anti-inflammatory activity. This study created a model in which CARAS was exacerbated by PM2.5 exposure, in BALB/c mice and explored the potential of bergapten as a therapeutic agent. The bergapten medication increased ovalbumin (OVA)-specific immunoglobulin (Ig) G2a level in serum and decreased OVA-specific IgE and IgG1 expression. Clinical nasal symptoms diminished significantly, with weakened inflammatory reaction in both the nasal mucosa and lungs. Furthermore, bergapten controlled the T helper (Th)1 to Th2 ratio by increasing cytokines associated with Th1-like interleukin (IL)-12 and interferon gamma and decreasing the Th2 cytokines IL-4, IL-5, and IL-13. Factors closely related to the balance between regulatory T cells and Th17 (such as IL-10, IL-17, Forkhead box protein P3, and retinoic-related orphan receptor gamma) were also regulated. Notably, pro-inflammatory cytokines IL-6, IL-1β, and tumor necrosis factor-alpha were reduced by bergapten, which suppressed the activation of both the signal transducer and activator of transcription 3 signaling pathway and the mitogen-activated protein kinase signaling pathway. Therefore, bergapten might have potential as a therapeutic agent for CARAS.

Topics: 5-Methoxypsoralen; Animals; Asthma; Cytokines; Disease Models, Animal; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Rhinitis, Allergic; STAT3 Transcription Factor; T-Lymphocytes, Regulatory

Pinellia ternata (Thunb.) Breit. (PT) has been demonstrated to be effective against the allergic airway inflammation (AAI) in clinical practices, especially in cold asthma (CA). Until now, the active ingredients, protective effect, and possible mechanism of PT against CA remain unknown.. The aim of this investigation was to examine the therapeutic impact and elucidate the underlying mechanism of PT on the AAI of CA.. The compositions of PT water extract were determined via the UPLC-Q-TOF-MS/MS. The ovalbumin (OVA) and cold-water baths were used to induce CA in female mice. Morphological characteristic observations, expectorant effect, bronchial hyperreactivity (BHR), excessive mucus secretion, and inflammatory factors were used to uncover the treatment effect of PT water extract. In addition, the mucin 5AC (MUC5AC) mRNA and protein levels and the aquaporin 5 (AQP5) mRNA and protein levels were detected via qRT-PCR, immunohistochemistry (IHC), and western blotting. Moreover, the protein expressions associated with the TLR4, NF-κB, and NLRP3 signaling pathway were monitored by western blot analysis.. Thirty-eight compounds were identified from PT water extract. PT showed significant therapeutic effects on mice with cold asthma in terms of expectorant activity, histopathological changes, airway inflammation, mucus secretion, and hyperreactivity. PT exhibited good anti-inflammatory effects in vitro and in vivo. The expression levels of MUC5AC mRNA and protein decreased significantly, while AQP5 expression levels increased significantly in the lung tissues of mice after administration with PT as compared to mice induced by CA. Furthermore, the protein expressions of TLR4, p-iκB, p-p65, IL-1β, IL-18, NLRP3, cleaved caspase-1, and ASC were markedly reduced following PT treatment.. PT attenuated the AAI of CA by modulating Th1- and Th2-type cytokines. PT could inhibit the TLR4-medicated NF-kB signaling pathway and activate the NLRP3 inflammasome to reduce CA. This study provides an alternative therapeutic agent of the AAI of CA after administration with PT.

Topics: Animals; Asthma; Expectorants; Female; Inflammation; Lung; Mice; NF-kappa B; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Pinellia; RNA, Messenger; Signal Transduction; Tandem Mass Spectrometry; Toll-Like Receptor 4

Pi-Pa-Run-Fei-Tang (PPRFT) is an empirical TCM prescription for treating asthma. However, the underlying mechanisms of PPRFT in asthma treatment have yet to be elucidated. Recent advances have revealed that some natural components could ameliorate asthma injury by affecting host metabolism. Untargeted metabolomics can be used to better understand the biological mechanisms underlying asthma development and identify early biomarkers that can help advance treatment.. The aim of this study was to verification the efficacy of PPRFT in the treatment of asthma and to preliminarily explore its mechanism.. A mouse asthma model was built by OVA induction. Inflammatory cell in BALF was counted. The level of IL-6, IL-1β, and TNF-α in BALF were measured. The levels of IgE in the serum and EPO, NO, SOD, GSH-Px, and MDA in the lung tissue were measured. Furthermore, pathological damage to the lung tissues was detected to evaluate the protective effects of PPRFT. The serum metabolomic profiles of PPRFT in asthmatic mice were determined by GC-MS. The regulatory effects on mechanism pathways of PPRFT in asthmatic mice were explored via immunohistochemical staining and western blotting analysis.. PPRFT displayed lung-protective effects through decreasing oxidative stress, airway inflammation, and lung tissue damage in OVA-induced mice, which was demonstrated by decreasing inflammatory cell levels, IL-6, IL-1β, and TNF-α levels in BALF, and IgE levels in serum, decreasing EPO, NO, and MDA levels in lung tissue, elevating SOD and GSH-Px levels in lung tissue and lung histopathological changes. In addition, PPRFT could regulate the imbalance in Th17/Treg cell ratios, suppress RORγt, and increase the expression of IL-10 and Foxp3 in the lung. Moreover, PPRFT treatment led to decreased expression of IL-6, p-JAK2/Jak2, p-STAT3/STAT3, IL-17, NF-κB, p-AKT/AKT, and p-PI3K/PI3K. Serum metabolomics analysis revealed that 35 metabolites were significantly different among different groups. Pathway enrichment analysis indicated that 31 pathways were involved. Moreover, correlation analysis and metabolic pathway analysis identified three key metabolic pathways: galactose metabolism; tricarboxylic acid cycle; and glycine, serine, and threonine metabolism.. This research indicated that PPRFT treatment not only attenuates the clinical symptoms of asthma but is also involved in regulating serum metabolism. The anti-asthmatic activity of PPRFT may be associated with the regulatory effects of IL-6/JAK2/STAT3/IL-17 and PI3K/AKT/NF-κB mechanistic pathways.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Immunoglobulin E; Interleukin-17; Interleukin-6; Lung; Lung Injury; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; Superoxide Dismutase; T-Lymphocytes, Regulatory; Tumor Necrosis Factor-alpha

There is now sufficient evidence to support an inverse association between helminth infection and secreted products with allergic/autoimmune disorders. Accordingly, several experimental studies have shown that Echinococcus granulosus infection and hydatid cyst compounds are able to suppress immune responses in allergic airway inflammation. This is the first study on effects of somatic antigens of E. granulosus on chronic allergic airway inflammation in BALB/c mice. Mice in OVA group were intraperitoneally (IP) sensitized with OVA/Alum. Subsequently, were challenged by nebulizing of OVA 1%. The treatment groups received somatic antigens of protoscoleces on the specified days. Mice in PBS group were received PBS in both sensitization and challenge. The effects of somatic products on development of chronic allergic airway inflammation were evaluated by examining histopathological changes, the recruitment of inflammatory cells in the bronchoalveolar lavage, cytokines production in the homogenized lung tissue, and total antioxidant capacity in serum. Our findings show that the co-administration of somatic antigens of protoscoleces simultaneously with the development of asthma intensifies allergic airway inflammation. The identification of effective components involved in exacerbation of allergic airway inflammation manifestations will be a crucial approach to understanding the mechanism of these interactions.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Disease Progression; Echinococcosis; Echinococcus granulosus; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Asthma is a chronic inflammatory disease of the airways with a high prevalence. Asthma has a complex pathophysiology and about 5-10% of patients are not fully responsive to the currently available treatments. The aim of this study is to investigate the involvement of NF-κB in the effects of fenofibrate on a mouse model of allergic asthma.. A total of 49 BALB/c mice were randomly distributed into 7 groups (n = 7). Allergic asthma model was created by administering i.p. injections of ovalbumin on days 0, 14 and 21, followed by provocation with inhaled ovalbumin on days 28, 29 and 30. Fenofibrate was orally given in 3 different doses; 1, 10 and 30 mg/kg through days 21-30 of the experiment. On day 31, pulmonary function test using whole body plethysmography was performed. The mice were sacrificed 24 h later. Blood samples were obtained, and serum of each sample was separated for IgE determination. Bronchoalveolar lavage fluid (BALF) and lung tissues were collected to measure IL-5 and IL-13 levels. Nuclear extracts of lung tissues were employed to assess nuclear factor kappa B (NF-κB) p65 binding activity.. Enhanced Pause (Penh) values were significantly increased in ovalbumin-sensitized and challenged mice (p < 0.01). Administration of fenofibrate (10 and 30 mg/kg) resulted in improved pulmonary function as shown by significantly lower Penh values (p < 0.01). Interleukin (IL) - 5 and IL-13 levels in BALF and lung tissues and immunoglobulin E (IgE) levels in serum were significantly elevated in the allergic mice. IL-5 levels in the lung tissues of mice treated with 1 mg/kg fenofibrate (FEN1) group were significantly reduced (p < 0.01). BALF and lung tissue IL-5 and IL-13 levels in mice treated with 10 and 30 mg/kg fenofibrate, FEN10 and FEN30, respectively, were significantly diminished when compared to the ovalbumin-treated (OVA) group, whereas treatment with 1 mg/kg fenofibrate resulted in insignificant changes. IgE levels in the serum of FEN30 group mice have shown a prominent reduction (p < 0.01). NF-κB p65 binding activity was higher in mice sensitized and challenged with ovalbumin (p < 0.01). NF-κB p65 binding activity was significantly reduced in allergic mice treated with 30 mg/kg (p < 0.01) fenofibrate.. In this study, we showed that administration of 10 and 30 mg/kg fenofibrate effectively attenuated airway hyperresponsiveness and inflammation in a mouse model of allergic asthma, possibly through inhibition of NF-κB binding activity.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Fenofibrate; Hypersensitivity; Immunoglobulin E; Interleukin-13; Interleukin-5; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Long noncoding RNAs (lncRNAs) are involved in gene transcription and pathophysiological processes of human diseases. Multiple lncRNAs have been shown to play important roles in the occurrence and development of asthma. This study aimed to explore the role of a novel lncRNA, lncRNA-AK007111, in asthma. Overexpression of lncRNA-AK007111 was induced in a mouse model of asthma via viral transfection, followed by the collection of alveolar lavage fluid and lung tissue for the detection of relevant inflammatory factors and pathological analysis of lung sections. Pulmonary resistance and respiratory dynamic compliance were measured using an animal pulmonary function analyzer. The number of mast cells sensitized by immunofluorescence was detected at the cellular level. The degree of degranulation of lncRNA-AK007111 after its knockdown was determined by detecting the level of β-hexosaminidase that was released and quantifying IL-6 and TNF-α using ELISA in a model of RBL-2H3 cells activated by immunoglobulin E plus antigen. Finally, we observed the migration ability of mast cells under a microscope. The results showed that in ovalbumin-sensitized mice, the upregulation of lncRNA-AK007111 promoted the infiltration of inflammatory cells in lung tissue, increased the number of total cells, eosinophils, and mast cells, upregulated IL-5 and IL-6 levels, and increased airway hyper-reactivity. Downregulation of lncRNA-AK007111 decreased the degranulation ability of IgE/Ag-activated mast cells and inhibited the expression of IL-6 and TNF-α; moreover, the migration ability of mast cells was significantly weakened. In conclusion, our study revealed that lncRNA-AK007111 plays an important role in asthma by modulating mast cell-related functions.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humans; Immunoglobulin E; Inflammation; Interleukin-6; Lung; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Long Noncoding; Tumor Necrosis Factor-alpha

Reduction of histone deacetylase (HDAC) 2 expression and activity may contribute to amplified inflammation in patients with severe asthma. Connective tissue growth factor (CTGF) is a key mediator of airway fibrosis in severe asthma. However, the role of the HDAC2/Sin3A/methyl-CpG-binding protein (MeCP) 2 corepressor complex in the regulation of CTGF expression in lung fibroblasts remains unclear.. The role of the HDAC2/Sin3A/MeCP2 corepressor complex in endothelin (ET)-1-stimulated CTGF production in human lung fibroblasts (WI-38) was investigated. We also evaluated the expression of HDAC2, Sin3A and MeCP2 in the lung of ovalbumin-induced airway fibrosis model.. HDAC2 suppressed ET-1-induced CTGF expression in WI-38 cells. ET-1 treatment reduced HDAC2 activity and increased H3 acetylation in a time-dependent manner. Furthermore, overexpression of HDAC2 inhibited ET-1-induced H3 acetylation. Inhibition of c-Jun N-terminal kinase, extracellular signal-regulated kinase, or p38 attenuated ET-1-induced H3 acetylation by suppressing HDAC2 phosphorylation and reducing HDAC2 activity. Overexpression of both Sin3A and MeCP2 attenuated ET-1-induced CTGF expression and H3 acetylation. ET-1 induced the disruption of the HDAC2/Sin3A/MeCP2 corepressor complex and then prompted the dissociation of HDAC2, Sin3A, and MeCP2 from the CTGF promoter region. Overexpression of HDAC2, Sin3A, or MeCP2 attenuated ET-1-stimulated AP-1-luciferase activity. Moreover, Sin3A- or MeCP2-suppressed ET-1-induced H3 acetylation and AP-1-luciferase activity were reversed by transfection of HDAC2 siRNA. In an ovalbumin-induced airway fibrosis model, the protein levels of HDAC2 and Sin3A were lower than in the control group; however, no significant difference in MeCP2 expression was observed. The ratio of phospho-HDAC2/HDAC2 and H3 acetylation in the lung tissue were higher in this model than in the control group. Overall, without stimulation, the HDAC2/Sin3A/MeCP2 corepressor complex inhibits CTGF expression by regulating H3 deacetylation in the CTGF promoter region in human lung fibroblasts. With ET-1 stimulation, the HDAC2/Sin3A/MeCP2 corepressor complex is disrupted and dissociated from the CTGF promoter region; this is followed by AP-1 activation and the eventual initiation of CTGF production.. The HDAC2/Sin3A/MeCP2 corepressor complex is an endogenous inhibitor of CTGF in lung fibroblasts. Additionally, HDAC2 and Sin3A may be of greater importance than MeCP2 in the pathogenesis of airway fibrosis.

Topics: Asthma; Co-Repressor Proteins; Connective Tissue Growth Factor; Endothelin-1; Fibroblasts; Histone Deacetylase 2; Humans; Luciferases; Lung; Ovalbumin; Pulmonary Fibrosis; Transcription Factor AP-1

Endocrine disrupting chemicals have been known to contribute to the aggravation of inflammatory diseases including asthma. We aimed to investigate the effects of mono-n-butyl phthalate (MnBP) which is one of the representing phthalates, and its antagonist in an eosinophilic asthma mouse model. BALB/c mice were sensitized by intraperitoneal injection of ovalbumin (OVA) with alum and followed by three nebulized OVA challenges. MnBP was administered through drinking water administration throughout the study period, and its antagonist, apigenin, was orally treated for 14 days before OVA challenges. Mice were assessed for airway hyperresponsiveness (AHR), differential cell count and type 2 cytokines in bronchoalveolar lavage fluid were measured in vivo. The expression of the aryl hydrocarbon receptor was markedly increased when MnBP was administered. MnBP treatment increased AHR, airway inflammatory cells (including eosinophils), and type 2 cytokines following OVA challenge compared to vehicle-treated mice. However, apigenin treatment reduced all asthma features, such as AHR, airway inflammation, type 2 cytokines, and the expression of the aryl hydrocarbon receptor in MnBP-augmented eosinophilic asthma. Our study suggests that MnBP exposure may increase the risk of eosinophilic inflammation, and apigenin treatment may be a potential therapy for asthma exacerbated by endocrine-disrupting chemicals.

Topics: Animals; Apigenin; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Aryl Hydrocarbon

Histone deacetylase inhibitors (HDACi) are novel class of drugs as they are involved in post translational modification of several proteins involved in signaling pathways related to asthma. HDACi have been reported to elicit protective effects on asthma but the signaling pathways associated with it have not been investigated much. Recently, we have demonstrated that intranasal administrations of Pan-HDAC inhibitors, sodium butyrate and curcumin, which have effectively reduced asthma severity via HDAC1 inhibition in Ovalbumin induced mouse model. Present study aimed to investigate possible pathways by which curcumin and sodium butyrate may minimize asthma pathogenesis via HDAC 1 inhibition. Balb/c mice were exposed (sensitized and challenged) with Ovalbumin to establish allergic asthma model followed by pretreatment of curcumin (5 mg/kg) and sodium butyrate (50 mg/kg) through intranasal route. Effects of curcumin and sodium butyrate on HIF-1α/VEGF signaling through activation of PI3K/Akt axis has been investigated using protein expressions followed by chromatin immunoprecipitation of BCL2 and CCL2 against HDAC1. Molecular docking analysis was also performed to investigate effects of curcumin and butyrate on mucus hypersecretion, goblet cell hyperplasia and airway hyperresponsiveness. Augmented expressions of HDAC-1, HIF-1α, VEGF, p-Akt and p-PI3K were observed in asthmatic group which was suppressed in both the treatments. NRF-2 level was significantly restored by curcumin and butyrate treatments. Protein expressions of p-p38, IL-5 and mRNA expressions of GATA-3 were also reduced in curcumin and butyrate treatment groups. Our findings suggest that curcumin and sodium butyrate may attenuate airway inflammation via down regulation of p-Akt/p-PI3K/HIF-1α/VEGF axis.

Topics: Animals; Antioxidants; Asthma; Butyric Acid; Curcumin; Histone Deacetylase Inhibitors; Mice; Molecular Docking Simulation; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Vascular Endothelial Growth Factor A

Childhood asthma is a major global health concern. ADP-ribosylation factor 6 (ARF6) is a low-molecular-weight GTPase; however, its role in childhood asthma remains unclear.. Ovalbumin (OVA)-challenged neonatal mice and transforming growth factor-β1 (TGF-β1)-induced BEAS-2B cells were used as. Upon OVA stimulation, ARF6 expression was upregulated in the lung tissue. Neonatal mice administered SehinH3 (an ARF6 inhibitor) exhibited improved pulmonary pathological injury, along with reduced inflammatory cell infiltration in the lungs and cytokine release in bronchial alveolar lavage fluid and serum (interleukin [IL]-3, IL-5, IL-13, IgE, and OVA-specific IgE). SehinH3 treatment restrained epithelial - mesenchymal transition (EMT) in the lungs of asthmatic mice, as evidenced by increased E-cadherin and decreased N-cadherin and α-smooth muscle actin expression. Different TGF-β1 exposures to BEAS-2B cells induced a time- and dose-dependent increase in ARF6 expression. Our study showed that ARF6 is associated with childhood asthma progression and may be positively regulated by E2F8. These results provide insight into the pathogenesis and treatment of childhood asthma.

Topics: ADP-Ribosylation Factor 6; Animals; Asthma; Disease Models, Animal; E2F Transcription Factors; Epithelial-Mesenchymal Transition; Immunoglobulin E; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Transforming Growth Factor beta1

Asthma is a persistent respiratory ailment that displays periodicity and is linked to the equilibrium of T cells. Several compounds obtained from Chinese herbal medicines display beneficial impacts on T cell regulation and the attenuation of inflammatory mediator synthesis. Schisandrin A, an active lignan derived from the Schisandra fruit, exhibits anti-inflammatory characteristics. In the present study, the network analysis conducted revealed that the nuclear factor-kappaB (NF-κB) signaling pathway is likely a prominent contributor to the anti-asthmatic effects of schisandrin A. In addition, it has been established that the inhibition of cyclooxygenase 2 (COX-2/PTGS2) is likely a significant factor in this process. The results of in vitro experiments have substantiated that schisandrin A can effectively lower the expression of COX-2 and inducible nitric oxide synthase (iNOS) in 16 HBE cells and RAW264.7 cells in a manner that is dependent on the dosage administered. It was able to effectively reduce the activation of the NF-κB signaling pathway while simultaneously improving the injury to the epithelial barrier function. Furthermore, an investigation utilizing immune infiltration as a metric revealed an inequity in Th1/Th2 cells and a surge in Th2 cytokines in asthma patients. In the OVA-induced asthma mice model, it was observed that schisandrin A treatment effectively suppressed inflammatory cell infiltration, reduced the Th2 cell ratio, inhibited mucus secretion, and prevented airway remodeling. To summarize, the administration of schisandrin A has been found to effectively alleviate the symptoms of asthma by impeding the production of inflammation, which includes reducing the Th2 cell ratio and improving the integrity of the epithelial barrier function. These findings offer valuable insights into the potential therapeutic applications of schisandrin A for the treatment of asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cyclooxygenase 2; Cytokines; Disease Models, Animal; Inflammation; Lignans; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Topics: Alkaloids; Animals; Anti-Allergic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Immunoglobulin E; Interleukin-13; Interleukin-6; Lung; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, IgE

The aim of the study was to discuss whether the anti-asthmatic effect of quercetin is related to periostin and the downstream molecular pathway of quercetin's anti-asthmatic effect.. We constructed asthmatic mice, sensitized by ovalbumin, and administrated different treatments into mice according to the experimental design. In this study, we mainly observed the inflammatory response, airway fibrosis, and airway hyperresponsiveness in asthmatic mice. Pathological stains (H&E, PAS, and Masson) were performed. We also detected the inflammation factors and fibrosis-related cytokines by enzyme-linked immunosorbent serologic assay. In addition, we also explored the level of periostin by enzyme-linked immunosorbent serologic assay and Western blot. At the same time, TGF-β1/Smad pathway was also determined by Western blot.. A high expression of periostin was found in asthmatic mice, and quercetin decreases periostin content in bronchoalveolar lavage fluid. Quercetin and OC-20 inhibit airway inflammation response, airway fibrosis, and airway hyperreactivity. Quercetin downregulated TGF-β1/Smad pathway in the lung tissues of asthmatic mice. Anti-asthma role of quercetin is related to periostin. Then deeper mechanical study revealed that inhibiting TGF-β1 could improve asthmatic symptoms, and quercetin exerted the protective effect on asthmatic mice through inhibition of TGF-β1/Smad pathway.. Quercetin provided a protective role against asthma via periostin, manifested by mild inflammatory infiltration, reduced goblet cell proliferation, and reduced airway fibrosis. TGF-β1/Smad pathway is an important transduction system, participating in the protective effect of quercetin on asthma.

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Fibrosis; Immunosorbents; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pulmonary Fibrosis; Quercetin; Transforming Growth Factor beta1

Tylophora indica (Burm. f.) Merr is a climbing perennial plant reported in Indian traditional system of medicine for its use in allergy and asthma. However, only few scientific studies have been performed in the past to validate its antiasthmatic potential.. The present study deals with investigation of airway smooth muscle relaxant and antiasthmatic potential of extract and subsequent fractions prepared from T. indica.. The most active fraction of T. indica leaves selected through bio-guided activity was subjected to liquid chromatography-mass spectrometry (LC-MS) analysis for chemical profiling. The binding affinity of identified compounds in fraction towards M. F-2 was found most effective in bioassay-guided fractionation. Characterization by LC-MS analysis revealed presence of five major bioactive compounds in F-2 that showed good docking interactions with M. These results confirm the traditional use of T. indica as an antiasthmatic agent which are evidenced through ex vivo, in silico and in vivo studies.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Guinea Pigs; Molecular Docking Simulation; Muscle, Smooth; Ovalbumin; Trachea; Tylophora

Asthma is a common illness with chronic airway inflammation. C1q/tumor necrosis factor (TNF)-related protein 3 (CTRP3) plays a vital role ininflammatory response, but its effect on asthma is imprecise. Herein, we analyzed the functions of CTRP3 in asthma.. The BALB/c mice were randomized into four groups: control, ovalbumin (OVA), OVA+vector, and OVA+CTRP3. The asthmatic mice model was established by OVA stimulation. Overexpression of CTRP3 was implemented by the transfection of corresponding adeno-associated virus 6 (AAV6). The contents of CTRP3, E-cadherin, N-cadherin, smooth muscle alpha-actin (α-SMA), phosphorylated (p)-p65/p65, transforming growth factor-beta 1 (TGFβ1), and p-Smad3/Smad3 were determined by Western blot analysis. The quantity of total cells, eosinophils, neutrophils, and lymphocytes in bronchoalveolar lavage fluid (BALF) was assessed by using a hemocytometer. The contents of tumor necrosis factor-α and interleukin-1β in BALF were examined by enzyme-linked immunesorbent serologic assay. The lung function indicators and airway resistance (AWR) were measured. The bronchial and alveolar structures were evaluated by hematoxylin and eosin staining and sirius red staining.. The CTRP3 was downregulated in mice of OVA groups; however, AAV6-CTRP3 treatment markedly upregulated the expression of CTRP3. Upregulation of CTRP3 diminished asthmatic airway inflammation by decreasing the number of inflammatory cells and the contents of proinflammatory factors. CTRP3 markedly lessened AWR and improved lung function in OVA-stimulated mice. Histological analysis found that CTRP3 alleviated OVA-induced airway remodeling in mice. Moreover, CTRP3 modulated NF-κB and TGFβ1/Smad3 pathways in OVA-stimulated mice.. CTRP3 alleviated airway inflammation and remodeling in OVA-induced asthmatic mice via regulating NF-κB and TGFβ1/Smad3 pathways.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Pediatric asthma is a common chronic disease of childhood with airway inflammation. Cyclic adenosine monophosphate response element binding protein (CREB) plays a significant role in the transcription of proinflammatory genes, but its role in pediatric asthma has remained unclear. Herein, we investigated the functions of CREB in pediatric asthma.. Eosinophils were purified from the peripheral blood of interleukin 5 (IL5) transgenic (IL5T) neonatal mice. The contents of CREB, long-chain fatty-acid-CoA ligase 4, transferrin receptor protein 1, ferritin heavy chain 1, and glutathione peroxidase 4 in eosinophils were examined by Western blot analysis. The viability of eosinophils, and the mean fluorescence intensity of Siglec F, C-C motif chemokine receptor 3 (CCR3), and reactive oxygen species were examined by flow cytometry. The concentration of iron in eosinophils was assessed by a commercial kit. The contents of malondialdehyde, glutathione, glutathione peroxidase, IL-5, and IL-4 were discovered by enzyme-linked-immunosorbent serologic assay. The C57BL/6 mice were randomly divided into four groups: sham, ovalbumin (OVA), OVA+Ad-shNC, and OVA+Ad-shCREB. The bronchial and alveolar structures were evaluated by hematoxylin and eosin staining. Leukocytes and eosinophils in the blood were measured using a HEMAVET 950.. The abundance of CREB in eosinophils was enhanced by CREB overexpression vector transfection, but reduced by short hairpin (sh)CREB transfection. Downregulation of CREB triggered the cell death of eosinophils. Knockdown of CREB could obviously contribute to ferroptosis of eosinophils. In addition, downregulation of CREB facilitated dexamethasone (DXMS, a type of glucocorticoid)-induced eosinophils death. Moreover, we established an asthma mouse model by OVA treatment. The CREB was upregulated in OVA group mice, but Ad-shCREB treatment obviously downregulated CREB level. Downregulation of CREB diminished OVA-induced asthmatic airway inflammation by reducing the number of inflammatory cells and the levels of proinflammatory factors. Downregulated CREB enhanced the anti-inflammatory effect of DXMS in OVA-induced mice.. Inhibition of CREB promoted the effect of glucocorticoids on airway inflammation in pediatric asthma through promoting ferroptosis of eosinophils.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Ferroptosis; Glucocorticoids; Inflammation; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin

Asthma is a common lung disease with increasing incidence and prevalence globally, thereby imposing a substantial global health and economic burden. Recently, studies have shown that Mitsugumin 53 (MG53) exhibits multiple biological functions and plays a protective role in a variety of diseases. However, the role of MG53 in asthma remained unknown; hence, in the present study we aimed to explore the functioning of MG53 in asthma.. Using ovalbumin and aluminum hydroxide adjuvant, an OVA-induced asthmatic animal model was constructed and administered with MG53. After establishing mice model, inflammatory cell counts and the levels of type 2 inflammatory cytokines were examined and histological staining of lung tissues were performed. The levels of key factors associated with the nuclear factor-κB (NF-κB) pathway were detected.. Asthmatic mice displayed a remarkable accumulation of white blood cells, neutrophils, macrophages, lymphocytes, and eosinophils in bronchoalveolar lavage fluid, compared to control mice. MG53 treatment lowered the number of these inflammatory cells in asthmatic mice. The level of type 2 cytokines in asthmatic mice was higher than that in control mice, and was lessened by MG53 intervention. In asthmatic mice, airway resistance was elevated, which was reduced by MG53 treatment. In addition, inflammatory cell infiltration and mucus secretion were aggravated in the lung tissues of asthmatic mice, and both were attenuated by MG53 intervention. The levels of phosphorylated p65 and phosphorylated inhibitor of nuclear factor kappa-B kinase were elevated in asthmatic mice, but were downregulated by MG53 supplement.. The aggravated airway inflammation was observed in asthmatic mice; however, MG53 treatment suppressed airway inflammation by targeting the NF-κB pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Lung; Membrane Proteins; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Allergic asthma, one of the most common types of asthma, is thought to be highly susceptible to respiratory viral infections; however, its pathological mechanism needs to be elucidated. Recent studies have found impaired T-cell function in asthmatic mice. Therefore, we aimed to investigate the way by which asthma induction affects T-cell exhaustion in the lungs and assess the relationship between T-cell exhaustion and influenza viral infection.. Chronic allergic asthma mice were induced by intranasal injection of ovalbumin for 6 weeks and asthmatic features and T cell populations in lung or airway were assessed. To determine the influenza virus susceptibility, control and asthma mice were challenged with the human influenza virus strain A/Puerto Rico/8/1934 H1N1 and evaluated the survival rate, lung damage, and virus titer.. Six weeks of OVA sensitization and challenge successfully induced chronic allergic asthma in a mouse model showing significant increase of sera IgE level and broncho-pathological features. A significant decrease in interferon-γ-producing T-cell populations and an increase in exhausted T-cell populations in the lungs of OVA-induced asthmatic mice were observed. Asthmatic mice were more susceptible to influenza virus infection than control mice showing lower survival rate and higher virus titer in lung, and a positive correlation existed between T-cell exhaustion in the lung and virus titer.. Asthma induction in mice results in the exhaustion of T-cell immunity, which may contribute to the defective capacity of viral protection. This study demonstrates a correlation between asthma conditions and viral susceptibility by investigating the functional characteristics of T-cells in asthma. Our results provide insights into the development of strategies to overcome the dangers of respiratory viral disease in patients with asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humans; Influenza A Virus, H1N1 Subtype; Influenza, Human; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; T-Cell Exhaustion

Allergic asthma is caused by chronic inflammation and hyper-responsiveness of the airway and is thought to be mediated by adaptive T helper type 2 (Th2)-driven immunity. However, recent studies have demonstrated that neuropeptide calcitonin gene-related peptide (CGRP)-mediated activation of group 2 innate lymphoid cells (ILC2s) may contribute to the development of asthma pathogenesis. Here, we investigated the therapeutic effects of the systemic administration of rimegepant, a CGRP receptor antagonist, on allergic asthma. Hyperplasia of CGRP-immunoreactive pulmonary neuroendocrine cells (PNECs) was observed in ovalbumin (OVA)-induced asthmatic mice. Concomitant with this, we observed an increase in the content of total lung CGRP. Upon antigen challenge, the concentration of plasma CGRP was transiently upregulated, whereas CGRP immunoreactivity within PNECs was intensively downregulated, suggesting that PNECs were the most likely source of CGRP. When rimegepant was administered according to CGRP kinetics, it suppressed asthma phenotypes, including airway hyper-responsiveness, infiltration of inflammatory cells in bronchoalveolar lavage fluid (BALF), hyperplasia of mucus-producing cells, and production of the Th2 cytokine IL-5. Moreover, we observed a decrease in the number of ILC2s and their capacity for IL-5 release in the presence of IL-33 in rimegepant-treated mice. In the allergic asthma model, rimegepant suppressed the activation of ILC2s mediated by PNEC-derived CGRP and subsequently impaired adaptive Th2-driven immunity, which ameliorated asthmatic phenotypes. Thus, an anti-CGRP signal strategy to target ILC2 will be a novel and attractive approach for treating allergic asthma that is refractory to other treatments.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Calcitonin Gene-Related Peptide; Calcitonin Gene-Related Peptide Receptor Antagonists; Cytokines; Down-Regulation; Hyperplasia; Immunity, Innate; Interleukin-5; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin

There is evidence that IL-22 and IL-17 participate in the pathogenesis of allergic asthma. To investigate the role of IL-22, we used IL-22 deficient mice (IL-22 KO) sensitized and challenged with ovalbumin (OVA) and compared with wild type (WT) animals exposed to OVA. IL-22 KO animals exposed to OVA showed a decreased number and frequency of eosinophils, IL-5 and IL-13 in the airways, reduced mucus production and pulmonary inflammation. In addition, IL-22 KO animals exhibited a decreased percentage and number of lung CD11c

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Comorbidity; Disease Models, Animal; Eosinophils; Interleukin-17; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Astragalus propinquus; Immunoglobulin E; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts

Asthma is a chronic inflammatory disorder in airways with typical pathologic features of airflow limitation, airway inflammation and remodeling. Icariside II (IS), derived from herbal medicine Herba Epimedii, exerts an anti-inflammatory property. However, underlying mechanisms with specifically targeted molecular expression by IS in asthma have not been fully understood, and whether IS could inhibit remodeling and EMT still remains unclear.. The study aimed to clarify therapeutic efficacy of IS for attenuating airway inflammation and remodeling in asthma, and illustrate IS-regulated specific pathway and target proteins through TMT-based quantitative proteomics.. Murine model of chronic asthma was constructed with ovalbumin (OVA) sensitization and then challenge for 8 weeks. Pulmonary function, leukocyte count in bronchoalveolar lavage fluid (BALF), lung histopathology, inflammatory and fibrotic cytokines, and markers of epithelial-mesenchymal transition (EMT) were evaluated. TMT-based quantitative proteomics were performed on lung tissues to explore IS-regulated proteins.. IS contributed to alleviative airway hyperresponsiveness (AHR) evidenced by declined R. The study demonstrated IS could ameliorate AHR, airway inflammation, remodeling and EMT in OVA-induced chronic asthma mice. Our research was the first to reveal that inhibition of LAMP2, CTSD and CTSS expression in autophagy contributed to the therapeutic efficacy of IS to asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Proteomics

Asthma is a chronic inflammatory disease with a high morbidity rate in children and significantly impacts their healthy growth. It is reported that Th2 cell-mediated airway inflammation and activated oxidative stress are involved in the pathogenesis of asthma. S14G-humanin (HNG) is a derivative of Humanin with higher activity. The present study proposes to explore the potential treating property of HNG on asthma. An asthma model was constructed in mice using ovalbumin (OVA), the mice were treated with 2.5 mg/kg and 5 mg/kg HNG for 16 days. Dramatically increased lung weight index, elevated number of monocytes, eosinophils, and neutrophils, promoted production of Th2 cytokines including interleukin-4 (IL-4), interleukin-5 (IL-5), and interleukin-13 (IL-13), and severe histological pathology were observed in OVA-challenged mice, all of which were extremely alleviated by 2.5 mg/kg and 5 mg/kg HNG. Furthermore, the increased malondialdehyde (MDA) level and declined superoxide dismutase (SOD) activity in OVA-challenged mice were abolished by 2.5 mg/kg and 5 mg/kg HNG. Lastly, the upregulated TLR4, p-NF-κB p65, and early growth response 1 (Egr-1) in lung tissues of OVA-challenged mice were pronouncedly downregulated by 2.5 mg/kg and 5 mg/kg HNG. Collectively, our data suggested that HNG ameliorated airway inflammation in asthma partially due to NF-κB and Egr-1-mediated responses.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Toll-Like Receptor 4

Obesity-related asthma is a kind of nonallergic asthma with excessive neutrophil infiltration in the airways. However, the underlying mechanisms have been poorly elucidated. Among the adipokines related to obesity, leptin is related to the inflammatory response. However, little is understood about how leptin acts on the leptin receptor (obR) in neutrophilic airway inflammation in obesity-associated asthma. We explored the inflammatory effects of leptin/obR signaling in an obesity-related neutrophilic airway inflammation mouse model.. We established a neutrophilic airway inflammation mouse model using lipopolysaccharide (LPS)/ovalbumin (OVA) sensitization and OVA challenge (LPS + OVA/OVA) in lean, obese, or db/db (obR deficiency) female mice. Histopathological, bronchoalveolar lavage fluid (BALF) inflammatory cell, and lung inflammatory cytokine analyses were used to analyze airway inflammation severity. Western blotting, flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR), and enzyme-linked immunosorbent assay (ELISA) were used to evaluate the underlying mechanisms. In vitro bone marrow-derived macrophage (BMDM) and bone marrow-derived neutrophil experiments were performed.. We found that the serum leptin level was higher in obese than in lean female mice. Compared to LPS/OVA + OVA-treated lean female mice, LPS/OVA + OVA-treated obese female mice had higher peribronchial inflammation levels, neutrophil counts, Th1/Th17-related inflammatory cytokine levels, M1 macrophage polarization levels, and long isoform obR activation, which could be decreased by the obR blockade (Allo-Aca) or obR deficiency, suggesting a critical role of leptin/obR signaling in the pathogenesis of obesity-related neutrophilic airway inflammation in female mice. In in vitro experiments, leptin synergized with LPS/IFN-γ to promote the phosphorylation of the long isoform obR and JNK/STAT3/AKT signaling pathway members to increase M1 macrophage polarization, which was reversed by Allo-Aca. Moreover, leptin/obR-mediated M1 macrophage activity significantly elevated CXCL2 production and neutrophil recruitment by regulating the JNK/STAT3/AKT pathways. In clinical studies, obese patients with asthma had higher serum leptin levels and M1 macrophage polarization levels in induced sputum than non-obese patients with asthma. Serum leptin levels were positively correlated with M1 macrophage polarization levels in patients with asthma.. Our results demonstrate leptin/obR signaling plays an important role in the pathogenesis of obesity-related neutrophilic airway inflammation in females by promoting M1 macrophage polarization.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Inflammation; Leptin; Lipopolysaccharides; Lung; Macrophages; Mice; Mice, Inbred BALB C; Obesity; Ovalbumin; Proto-Oncogene Proteins c-akt; Receptors, Leptin; Signal Transduction

The transient receptor potential (TRP) channels regulate physiological and pathological processes. Changes in their activity and sensitivity may be involved in the pathophysiology of asthma. The present study investigates the effect of an inhaled TRPV4 channel blocker HC-067047 in an experimental guinea pig model of ovalbumin-induced allergic asthma. We monitored the effect of 50 nM, 100 nM, and 150 nM HC-067047 concentrations on airway defense reflexes in vivo and tracheal smooth muscle contractility in vitro. The anti-inflammatory action of HC-067047 was investigated by analysis of chronic inflammation markers from lung homogenates. The results suggest that HC-067047 can suppress airway defense reflexes in vivo and acetylcholine-induced contractility in vitro. Immunological analysis revealed that TRPV4 channel blockade leads to a decrease in the levels of inflammatory cytokines. An effect on airway defence reflexes and airway inflammation was observed using tested concentrations (50 mM, 100 mM, 150 mM) of HC-067047. The effects of HC-067047 on both airway defense reflexes and inflammation underline the role of TRPV4 channels in asthma and uncover therapeutic targets for developing innovative drugs in asthma therapy.

Topics: Animals; Asthma; Disease Models, Animal; Guinea Pigs; Inflammation; Lung; Muscle, Smooth; Ovalbumin; TRPV Cation Channels

Increasing attention has been paid to the potential impact of microplastics (MPs) pollution on human health. MPs and phthalates coexist in the environment, however, the effects of exposure to MPs alone or to a combination of di-(2-ethylhexyl) phthalate (DEHP) and MPs on allergic asthma are unclear. This study investigates the effects of exposure to polystyrene microplastics (PS-MPs) or co-exposure with DEHP, on allergic asthma, and the underlying molecular mechanisms. We established an allergic asthma model using ovalbumin, and mice were exposed to PS-MPs (5 mg/kg bw/day) alone, or combined with DEHP (0.5, 5 mg/kg bw/day), for 28 days. The results showed that in the presence of ovalbumin (OVA) sensitization, exposure to PS-MPs alone slightly affected airway inflammation, and airway hyperresponsiveness, while co-exposure to PS-MPs and DEHP caused more significant damage. Co-exposure also induced more oxidative stress and Th2 immune responses, and activation of the TRPA1 and p38 MAPK pathways. The aggravation of asthmatic symptoms induced by co-exposure to PS-MPs and DEHP were inhibited by blocking TRPA1 ion channel or p38 MAPK pathway. The results demonstrated that co-exposure to PS-MPs and DEHP exacerbates allergic asthma, by exacerbating oxidative stress and inflammatory responses, and activating the TRPA1-p38 MAPK pathway.

Topics: Animals; Asthma; Diethylhexyl Phthalate; Mice; Microplastics; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Plastics; Polystyrenes; TRPA1 Cation Channel

Previous studies have reported a correlation between the dysregulation of intestinal microbiota and the occurrence of asthma. This study aimed to investigate the effect of probiotic Lactobacillus rhamnosus 76 (LR76) on ovalbumin (OVA)-allergic mice and the mechanism of LR76 affecting mucus secretion in asthma. OVA-allergic mice were supplemented with LR76, and 16HBE cells induced by interleukin-13 (IL-13) were treated with LR76 supernatant (LR76-s) to observe the effect of LR76. In OVA-sensitized mice, LR76 alleviated the inflammatory cell infiltration in lung tissue and reduced the inflammatory cell counts of BALF. The expression level of mRNA, including Il4, Il5, Il13, Il25, Tgfb1, Il10, and Ifng, was decreased in the lung tissue of mice in the LR76 group compared with the OVA group. MUC5AC expression was down-regulated, while SCGB1A1 was up-regulated in the lung tissue of OVA-allergic mice after being supplemented with LR76 and in 16HBE cells induced by IL-13 after incubating with LR76-s. LR76 and LR76-s down-regulated the expression of proteins, including STAT6, p-STAT6, and SPDEF, and mRNA of STAT6 and SPDEF. In conclusion, LR76 alleviated airway inflammation and Th2 response in OVA-allergic mice and improved the mucus secretion of mouse lung tissue and 16HBE cells in the asthma model by down-regulating STAT6/SPDEF pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humans; Hypersensitivity; Inflammation; Interleukin-13; Lacticaseibacillus rhamnosus; Lung; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; RNA, Messenger; Transcription Factors

Asthma is a common chronic allergic disease that affects a significant percentage of the world's population. Niosomes are nanoparticles consisting of non-ionic surfactants that can be used for drug delivery. This research was designed to investigate the impacts of inhalation of simple and niosomal forms of myrtenol against adverse consequences of asthma in rats. Asthma induction was performed via injection of ovalbumin, followed by its inhalation. Niosomes were created by a heating protocol, and their physicochemical features were evaluated. Forty-nine male Wistar rats were allotted into 7 groups (n=7 each): Control (CTL), vacant niosome (VN), Asthma, Asthma+VN, Asthma+SM (simple myrtenol), Asthma+NM (niosomal myrtenol), and Asthma+B (budesonide). Lung remodeling, serum immunoglobulin E (IgE), inflammatory  and cytokines, and antioxidant factors in the lung tissue and bronchoalveolar fluid (BALF), as well as), were evaluated. The results showed that myrtenol-loaded niosomes had appropriate encapsulation efficiency, kinetic release, size, and zeta potential. The thickness of the epithelial cell layer in the lungs, as well as cell infiltration, fibrosis, IgE, reactive oxygen species, interleukin (IL)-6, and tumor nuclear factor alpha (TNF-α) levels, decreased significantly. In contrast, superoxide dismutase and glutathione peroxide activity increased significantly in the serum and BALF of the treated groups. The niosomal form of myrtenol revealed a higher efficacy than simple myrtenol and was similar to budesonide in ameliorating asthma indices.  Inhalation of simple and niosomal forms of myrtenol improved the detrimental changes in the asthmatic lung. The niosomal form induced more prominent anti-asthmatic effects comparable to those of budesonide.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Budesonide; Cytokines; Disease Models, Animal; Immunoglobulin E; Interleukin-6; Liposomes; Lung; Male; Ovalbumin; Rats; Rats, Wistar

Combined allergic rhinitis and asthma syndrome (CARAS) is an allergic airway inflammatory disorder orchestrated by the type 2 immune response. The close gut-lung relationship has been described, however, the effect of gut-modulating agents such as probiotics in allergic airway disorder is unclear. Thus, the goal of this study was to evaluate theLimosilactobacillus fermentumsupplementation in animals with CARAS. Therefore, BALB/c mice were ovalbumin (OVA) -sensitized and -challenged after being supplemented withL. fermentum. Animals, previously probiotic supplemented, showed a decrease (p < 0.05) of inflammatory cell migration, mainly eosinophil, into the nasal (NALF) and the bronchoalveolar (BALF) fluids as well as reduction of the allergic signs such as sneezing, nasal rubbings, and nasal hyperreactivity induced by histamine as compared with non-supplemented animals. In the systemic context,L. fermentumreduced eosinophilia and the serum levels of OVA-specific IgE. The altered histological aspects of nasal and lung tissues of animals with CARAS were effectively ameliorated byL. fermentum. In the BALF, the immunomodulatory effect was due to the decreasing of type 2 and 3 cytokines (IL-4, IL-13, IL-5 and IL-17A) dependent on type 1 (IFN-γ) and Treg (IL-10) cytokine increasing. Indeed,L. fermentumimproved the FOXP3 activation. Additionally, these effects correlate with the amplification of the gut response as increasing short-chain fatty acids (SCFAs) levels, gut epithelium barrier (ZO-1) maintenance, and colon tissue integrity. These data pointed out that animals' probiotic supplemented presented immunomodulatory responses in CARAS experimental model by activating the intracellular transduction signal underlying the IL-10 gene transcription.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Forkhead Transcription Factors; Immunity; Interleukin-10; Limosilactobacillus fermentum; Mice; Mice, Inbred BALB C; Ovalbumin; Rhinitis, Allergic

As particulate matter (PM) poses an increasing risk, research on its correlation with diseases is active. However, researchers often use their own PM, making it difficult to determine its components. To address this, we investigated the effects of PM with known constituents on BEAS-2B cells, examining cytokine levels, reactive oxygen species ROS production, DNA damage, and MAPK phosphorylation. Additionally, we evaluated the effects of PM on normal and OVA-induced asthmatic mice by measuring organ weight, cytokine levels, and inflammatory cells in bronchoalveolar lavage fluid, and examining histological changes. PM markedly increased levels of IL-6, GM-CSF, TNF-α, ROS, nitric oxide, and DNA damage, while surprisingly reducing IL-8 and MCP-1. Moreover, PM increased MAPK phosphorylation and inhibited mTOR and AKT phosphorylation. In vivo, lung and spleen weights, IgE, OVA-specific IgE, IL-4, IL-13, total cells, macrophages, lymphocytes, mucus generation, and LC3II were higher in the asthma group. PM treatment in asthmatic mice increased lung weight and macrophage infiltration, but decreased IL-4 and IL-13 in BALF. Meanwhile, PM treatment in the Nor group increased total cells, macrophages, lymphocytes, and mucus generation. Our study suggests that PM may induce and exacerbate lung disease by causing immune imbalance via the MAPK and autophagy pathways, resulting in decreased lung function due to increased smooth muscle thickness and mucus generation.

Topics: Animals; Asthma; Autophagy; Bronchoalveolar Lavage Fluid; Cytokines; Immunoglobulin E; Inflammation; Interleukin-13; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Reactive Oxygen Species

Asthma is a chronic inflammatory lung disease that causes respiratory difficulties. Black ginseng extract (BGE) has preventative effects on respiratory inflammatory diseases such as asthma. However, the pharmacological mechanisms behind the anti-asthmatic activity of BGE remain unknown. To investigate the anti-asthmatic mechanism of BGE, phorbol 12-myristate 13-acetate plus ionomycin (PMA/Iono)-stimulated mouse EL4 cells and ovalbumin (OVA)-induced mice with allergic airway inflammation were used. Immune cells (eosinophils/macrophages), interleukin (IL)-4, -5, -13, and serum immunoglobulin E (IgE) levels were measured using an enzyme-linked immunosorbent assay. Inflammatory cell recruitment and mucus secretion in the lung tissue were estimated. Protein expression was analyzed via Western blotting, including that of inducible nitric oxide synthase (iNOS) and the activation of protein kinase C theta (PKCθ) and its downstream signaling molecules. BGE decreased T helper (Th)2 cytokines, serum IgE, mucus secretion, and iNOS expression in mice with allergic airway inflammation, thereby providing a protective effect. Moreover, BGE and its major ginsenosides inhibited the production of Th2 cytokines in PMA/Iono-stimulated EL4 cells. In EL4 cells, these outcomes were accompanied by the inactivation of PKCθ and its downstream transcription factors, such as nuclear factor of activated T cells (NFAT), nuclear factor kappa B (NF-κB), activator of transcription 6 (STAT6), and GATA binding protein 3 (GATA3), which are involved in allergic airway inflammation. BGE also inhibited the activation of PKCθ and the abovementioned transcriptional factors in the lung tissue of mice with allergic airway inflammation. These results highlight the potential of BGE as a useful therapeutic and preventative agent for allergic airway inflammatory diseases such as allergic asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Inflammation; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Panax; Signal Transduction

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Fallopia japonica; Inflammation; Interleukin-33; Male; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Rhinitis, Allergic; Signal Transduction

Evidence suggest that the renin-angiotensin system (RAS) is activated in people with asthma, although its pathophysiological role is unclear. Angiotensin-converting enzyme 2 (ACE2) is the major enzyme that converts angiotensin II to angiotensin 1-7 (Ang-1-7), and is also known as a receptor of SARS-CoV-2. The current study was conducted to identify the change in RAS-related gene expression in airways of a murine asthma model.. The ovalbumin (OA)-sensitized mice were repeatedly challenged with aerosolized OA to induce asthmatic reaction. Twenty-four hours after the last antigen challenge, the main bronchial smooth muscle (BSM) tissues were isolated.. The KEGG pathway analysis of differentially expressed genes in our published microarray data revealed a significant change in the RAS pathway in the antigen-challenged mice. Quantitative RT-PCR analyses showed significant increases in the angiotensin II-generating enzymes (Klk1, Klk1b3 and Klk1b8) and a significant decrease in Ace2. Surprisingly, ELISA analyses revealed a significant increase in Ang-1-7 levels in bronchoalveolar lavage (BAL) fluids of the antigen-challenged animals, while no significant change in angiotensin II was observed. Application of Ang-1-7 to the isolated BSMs had no effect on their isometrical tension.. The expression of Ace2 was downregulated in the BSMs of OA-challenged mice, while Klk1, Klk1b3 and Klk1b8 were upregulated. Despite the downregulation of ACE2, the level of its enzymatic product, Ang-1-7, was increased in the inflamed airways, suggesting the existence of an unknown ACE2-independent pathway for Ang-1-7 production. The functional role of Ang-1-7 in the airways remains unclear.

Topics: Angiotensin II; Angiotensin-Converting Enzyme 2; Animals; Asthma; COVID-19; Down-Regulation; Gene Expression; Mice; Ovalbumin; Renin-Angiotensin System; SARS-CoV-2

Topics: Animals; Antioxidants; Asthma; Inflammation; Kelch-Like ECH-Associated Protein 1; Mice; Molecular Docking Simulation; NF-E2-Related Factor 2; Ovalbumin; Signal Transduction

Asthma is a chronic inflammatory disease around the world. Extracellular adenosine triphosphate works as a dangerous signal in responding to cellular stress, irritation, or inflammation. It has also been reported its association with the pathogenicity in asthma, with increased level in lungs of asthmatics. Pannexin-1 is one of the routes that contributes to the release of adenosine triphosphate form intracellular to extracellular. The aim of this study was to apply pannexin-1 peptide antagonist

Topics: Adenosine Triphosphate; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Connexins; Cytokines; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Nerve Tissue Proteins; Ovalbumin

Human rhinoviruses are known to predispose infants to asthma development during childhood and are often associated with exacerbations in asthma patients. MYADM epithelial expression has been shown to associate with asthma severity. The goal of this study was to determine if MYADM expression patterns were altered in asthma and/or rhinovirus infection and if increased MYADM expression is associated with increased asthma-associated factors.. Utilizing H1HeLa cells and differentiated primary human airway epithelial cells (AECs), we measured the expression of MYADM and inflammatory genes by qRT-PCR in the presence or absence of RV-1B infection or poly I:C treatment and with siRNA knockdown of MYADM. Expression of MYADM in the asthmatic lung was determined in the ovalbumin (ova)-challenged murine model.. MYADM expression was upregulated in the lungs from ova-treated mice and in particular on the subsurface vesicle membrane in airway epithelial cells. Upon infection with RV-1B, human AECs grown at an air-liquid interface had increased the MYADM expression predominantly detected in ciliated cells. We found that the presence of MYADM was required for expression of several inflammatory genes both in a resting state and after RV-1B or poly I:C treatments.. Our studies show that in a mouse model of asthma and during RV-1B infection of primary human AECs, increased MYADM expression is observed. In the mouse model of asthma, MYADM expression was predominantly on the luminal side of airway epithelial cells. Additionally, MYADM expression was strongly associated with increases in inflammatory genes, which may contribute to more severe asthma and RV-linked asthma exacerbations.

Topics: Animals; Antigens, Differentiation; Asthma; Disease Models, Animal; Enterovirus Infections; Humans; Infant; Mice; Ovalbumin; Poly I-C; Rhinovirus

To explore the mechanism of moxibustion in the treatment of asthmatic inflammation from the point of short-chain fatty acids (SCFAs) in rats with asthma.. A total of 48 SD rats (half male and half female) were randomly divided into 4 groups： normal, model, lung treatment and joint-treatment of lung and intestine (joint-treatment), with 12 rats in each group. The asthma model was made by subcutaneous (bilateral back and inguinal regions) and intraperitoneal injection of mixture solution of ovalbumin and aluminium hydroxide gel (on day 1 and 8) and followed by inhalation of atomized 1% ovalbumin (20 min from day 15, once daily for one week). Moxibustion was applied to bilateral "Feishu" (BL13) for rats of the lung treatment group or bilateral "Feishu" (BL13) and "Tianshu" (ST25) for rats of the joint treatment group. One hour after the intervention, the rats in the later three groups were separately given atomized 1% ovalbumin solution inhalation for 20 min. The treatment was conducted for 30 min, once daily for 14 consecutive days. At the end of the intervention, the percentage of inflammatory cells in blood was detected by biochemical method and histopathological changes of the lung were observed after H.E. staining. The inflammatory cells in the bronchoalveolar lavage fluid (BALF) were counted after Wright-Giemsa staining. The mRNA expressions of interleukin (IL)-4, IL-5, IL-13, IL-17, IL-33, leukotriene (LT), thymic stromal lymphopoietin (TSLP) and prostaglandin D2 (PGD2) were detected by real-time PCR, and the contents of SCFAs in rats' feces were detected by gas chromatography-mass spectrometry.. Relevant to the normal group, the model group had an obvious increase in the percentages of neutrophils, lymphocytes and eosinophils in the blood, the percentages of neutrophils and eosinophils in the BALF, and in the expression levels of PGD2, TSLP, LT, IL-4, IL-5, IL-13, IL-17 and IL-33 mRNAs in the lung tissues (. Joint treatment of asthma from the lung and intestine can better regulate the contents of intestinal SCFAs and alleviate the inflammatory response of asthmatic model rats, thus, intestinal SCFAs may be involved in the process of moxibustion in improving inflammatory response.

Topics: Acupuncture Points; Animals; Asthma; Fatty Acids, Volatile; Female; Interleukin-13; Interleukin-17; Interleukin-33; Interleukin-4; Interleukin-5; Intestines; Isobutyrates; Lung; Male; Moxibustion; Ovalbumin; Pneumonia; Propionates; Prostaglandin D2; Rats; Rats, Sprague-Dawley

Excretory/secretory products (ESPs) derived from helminths have been reported to effectively control allergic inflammation, which have better therapeutic prospects than live parasite infections. However, it remains unknown whether ESPs from schistosome eggs can protect against allergies, despite reports alleging that schistosome infection could alleviate disordered allergic inflammation.. In the present study, we investigated the protective effects of ESPs from Schistosoma japonicum eggs (ESP-SJE) on asthmatic inflammation. Firstly, we successfully established an allergic airway inflammation model in mice by alum-adjuvanted ovalbumin (OVA) sensitization and challenge. ESP-SJE were administered intraperitoneally on days -1 and 13 (before sensitization), on day 20 (before challenge), and on days 21-24 (challenge phase).. The results showed that ESP-SJE treatment significantly reduced the infiltration of inflammatory cells, especially eosinophils into the lung tissue, inhibited the production of the total and OVA-specific IgE during OVA-sensitized and -challenged phases, respectively, and suppressed the secretion of Th2-type inflammatory cytokines (IL-4). Additionally, ESP-SJE treatment significantly upregulated the regulatory T cells (Tregs) in the lung tissue during OVA challenge. Furthermore, using liquid chromatography-mass spectrometry analysis and Treg induction experiments in vitro, we might identify nine potential therapeutic proteins against allergic inflammation in ESP-SJE. The targets of these candidate proteins included glutathione S-transferase, egg protein CP422 precursor, tubulin alpha-2/alpha-4 chain, actin-2, T-complex protein 1 subunit beta, histone H₄, whey acidic protein core region, and molecular chaperone HtpG.. Taken together, the results discussed herein demonstrated that ESP-SJE could significantly alleviate OVA-induced asthmatic inflammation in a murine model, which might be mediated by the upregulation of Treg in lung tissues that may be induced by the potential modulatory proteins. Therefore, potential proteins in ESP-SJE might be the best candidates to be tested for therapeutic application of asthma, thus pointing out to a possible new therapy for allergic airway inflammation.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Egg Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Schistosoma japonicum

Pingchuan formula is a traditional Chinese herbal prescription for asthma, but its components and underlying mechanisms remain unclear. Here, we evaluated its anti-asthmatic actvity and regulatory effects on the gut microbiota in mice based on the traditional Chinese medicine Zang-Fu theory, which proposed the exterior-interior relationship between the lung and the large intestine.. Mouse model withovalbumin (OVA)-induced asthma was used to assess the protective effect of the water extract of Pingchuan formula (PC). The chemical compounds of PC and mouse serum metabolites were identified by Ultraperformance liquid chromatography-Q Exactive HF-X spectrometry. Gut microbiota was evaluated by 16 S rRNA gene sequencing. The gut microbiota was depleted with a broad-spectrum antibiotic mixture (Abx) to explore whether it plays a role in the protective effects of PC.. PC mainly contains phenols, flavonoids, alkaloids, carboxylic acids, and their derivatives. PC attenuated OVA-induced asthma in mice by alleviating inflammatory infiltration, indicated by decreased levels of IL-18, IL-6, IL-4, and Eotaxin in lung tissues. PC treatment altered the serum metabolites and affected the pyrimidine pathway. In addition, our results showed that acacetin and abscisic acid were the key serum metabolites PC treatment changed the composition of gut microbiota by increasing the relative abundance of Clostridia_UCG_014 and Akkermansia while decreasing Blautia, Barnesiella, and Clostridium_Ⅲ at the genus level. Importantly, the Abx treatment partly abolished the anti-asthmatic effect of PC.. We demonstrated that PC could alleviate OVA-induced asthma in mice and protect against inflammatory infiltration in lungs via modulating the serum metabolites and gut microbiota, thereby providing a new reference for the therapeutic effect of PC.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Drugs, Chinese Herbal; Gastrointestinal Microbiome; Mice; Ovalbumin

Secretion of translationally controlled tumor protein (TCTP) was found in body fluids during the late phase of allergic reactions, implicating TCTP in allergic diseases. Furthermore, blocking TCTP has been shown to be helpful in treating asthma and allergies in animal models. The objectives of this study were to produce anti-TCTP monoclonal antibodies (mAbs), test their ability to inhibit the cytokine-like function of dimeric TCTP (dTCTP) in vitro and to assess their therapeutic effects in a murine model of ovalbumin (OVA)-induced airway inflammation. We first verified the inhibitory effects of 4 anti-TCTP mAbs on dTCTP-induced secretion of IL-8 in BEAS-2B cells. To investigate the anti-inflammatory effect of anti-TCTP mAbs on allergic airway inflammation, we treated OVA-sensitized mice with anti-TCTP mAbs before OVA challenge. The changes in bronchoalveolar lavage fluid (BALF) cells, IL-4, IL-5, and IL-13 levels in both BALF and lung homogenates, plasma levels of OVA-specific IgE, and lung tissues were analyzed. We found that JEW-M449 anti-TCTP mAb bound to the flexible loop of TCTP and significantly inhibited dTCTP-induced IL-8 release, making it the most effective inhibitor in our study. We also found that treatment with JEW-M449 significantly reduced the infiltration of inflammatory cells and suppressed the OVA-induced upregulation of type 2 cytokines in both BALF and lung homogenates in a dose-dependent manner. In addition, JEW-M449 significantly attenuated the degree of goblet cell hyperplasia and mucus secretion. Our results demonstrate that specific targeting of the flexible loop of TCTP is a potent strategy for treating airway inflammatory diseases.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Inflammation; Interleukin-8; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Tumor Protein, Translationally-Controlled 1

Asthma is a respiratory inflammatory disease, and nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) is involved in the progression of respiratory diseases. However, the role of NOX4 in asthma remains unclear. In the present study, we aimed to explore the effects of NOX4 on airway remodeling and inflammation. NOX4 expression was measured using immunocytochemistry (IHC), western blot, and real-time PCR (qPCR). Lung tissues were stained using the H&E assay. ELISA was used to examine the levels of airway remodeling-related indicators, and qPCR was used to detect airway inflammatory factors. The results indicated that NOX4 is highly expressed in lung tissues, bronchoalveolar lavage fluid (BALF), and serum of OVA-treated mice. Inhibition of NOX4 alleviated OVA-induced airway remodeling and inflammation. Similarly, TGF-β1 was also upregulated in BALF and serum OVA-induced mice. Inhibition of TGF-β1 signaling also improved airway remodeling and inflammation induced by OVA. Moreover, the downregulation of NOX4 inactivated the TGF-β1-Smad2/3 pathway, and TGF-β1 decreased Smad2/3 expression. Moreover, inhibition of the TGF-β1 was enhanced, while TGF-β1 reversed the effects on airway remodeling and inflammation induced by NOX4 inhibition. Taken together, the downregulation of NOX4 improves airway remodeling and inflammation via inactivation of the TGF-β1-Smad2/3 pathway in asthma mice, suggesting that NOX4 may be a therapeutic target for asthma.

Topics: Airway Remodeling; Animals; Asthma; Down-Regulation; Inflammation; Mice; NADPH Oxidase 4; Ovalbumin; Transforming Growth Factor beta1

Neutrophilic airway inflammation is a challenge in asthma management and is associated with poor patient prognosis. Mucin 1 (MUC1), which contains a cytoplasmic tail (MUC1-CT), has been found to mediate glucocorticoid sensitivity in asthma; however, its role in modulating neutrophilic airway inflammation in asthma remains unknown.. Human-induced sputum cells were collected from healthy participants (n = 12), patients with mild-to-moderate asthma (n = 34), and those with severe asthma (n = 18). In vitro human lung bronchial 1 epithelial cell line (BEAS-2B) was transfected with small interfering RNA against MUC1 (MUC1-siRNA) and then stimulated by lipopolysaccharide (LPS), where some cells were pretreated with a TLR4 inhibitor (TAK-242). In vivo mouse model of asthmatic neutrophil airway inflammation was induced by ovalbumin (OVA)/LPS. Some groups were intraperitoneally injected with MUC1-CT inhibitor (GO-203) and/or TAK-242 .. The mRNA expression of MUC1 was downregulated in the induced sputum of patients with asthma and correlated with asthmatic neutrophilic airway inflammation. The mRNA expressions of TLR4, MyD88, nucleotide-binding oligomerization domain-like pyrin domain-containing protein 3 (NLRP3), caspase-1, interleukin (IL)-18, and IL-1β in induced sputum cells of patients with asthma were upregulated and related to the mRNA expression of MUC1. LPS activated the TLR4 pathway and NLRP3-mediated pyroptosis in BEAS-2B cells in vitro, which were significantly aggravated after MUC1-siRNA transfection. Furthermore, MUCl-CT interacted with TLR4, and the interaction between TLR4 and MyD88 was significantly increased after MUCl-siRNA transfection. Moreover, TAK-242 ameliorated TLR4/MyD88/nuclear factor kappa B (NF-κB) pathway activation, NLRP3 inflammasome-mediated pyroptosis, and neutrophilic inflammation exacerbated by MUC1 downregulation. GO-203 exacerbated TLR4/MyD88/NF-κB pathway activation in vivo, and NLRP3 inflammasome-mediated pyroptosis reduced in a mouse model of asthmatic neutrophil airway inflammation induced by OVA/LPS; these pathological changes were partially alleviated after TAK-242 application.. This study revealed that MUC1 downregulation plays an important role in asthmatic neutrophilic airway inflammation. MUC1-CT reduces NLRP3 inflammasome-mediated pyroptosis by inhibiting the activation of the TLR4/MyD88/NF-κB pathway, thereby attenuating neutrophil airway inflammation in patients with asthma.

Topics: Animals; Asthma; Humans; Inflammasomes; Inflammation; Lipopolysaccharides; Mice; Mucin-1; Myeloid Differentiation Factor 88; NF-kappa B; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Pyroptosis; RNA, Messenger; RNA, Small Interfering; Signal Transduction; Toll-Like Receptor 4

Despite recent investigations on the role of chitinase in asthma, its role in obesity-induced asthma has not been evaluated. Therefore, we investigated the roles of chitin, chitinase-1, and a chitinase-1 inhibitor (compound X, CPX) in a murine model.. We assigned C57BL/6 mice to the ovalbumin (OVA) model or obesity model group. In the OVA model, mice received intraperitoneal OVA twice within a 2-week interval and intranasal OVA for 3 consecutive days. Additionally, chitin was intranasally administered for 3 consecutive days, and CPX was intraperitoneally injected three times over 5 days. In the obesity model, a high-fat diet (HFD) was maintained for 13 weeks, and CPX was intraperitoneally injected eight times over 4 weeks.. In the OVA model, chitin aggravated OVA-induced airway hyper-responsiveness (AHR), increased bronchoalveolar lavage fluid (BALF) cell proliferation, increased fibrosis, and increased the levels of various inflammatory cytokines (including chitinase-1, TGF-β, TNF-α, IL-1 β, IL-6, IL-4, and IL-13). CPX treatment significantly ameliorated these effects. In the obesity model, HFD significantly increased AHR, BALF cell proliferation, fibrosis, and the levels of various inflammatory cytokines. Particularly, compared to the control group, the mRNA expression of chitinase, chitinase-like molecules, and other molecules associated with inflammation and the immune system was significantly upregulated in the HFD and HFD/OVA groups. Immunofluorescence analysis also showed increased chitinase-1 expression in these groups. CPX significantly ameliorated all these effects in this model.. This study showed that CPX can be an effective therapeutic agent in asthma, especially, obesity-induced and -aggravated asthma to protect against the progression to airway remodeling and fibrosis.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Chitin; Cytokines; Disease Models, Animal; Fibrosis; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Obesity; Ovalbumin

Allergic asthma is associated with chronic airway inflammation and progressive airway remodelling. The sclerotium of Lignosus rhinocerotis (Cooke) Ryvarden (Tiger Milk mushroom) is used traditionally to treat various illnesses, including asthma in Southeast Asia. This study was carried out to evaluate the effect of L. rhinocerotis extract (LRE) on airway inflammation and remodelling in a chronic model of asthma. The present study investigated the therapeutic effects of LRE on airway inflammation and remodelling in prolonged allergen challenged model in allergic asthma. Female Balb/C mice were sensitised using ovalbumin (OVA) on day 0 and 7, followed by OVA-challenged (3 times/week) for 2, 6 and 10 weeks. LRE (125, 250, 500 mg/kg) were administered by oral gavage one hour after every challenge. One group of mice were left untreated after the final challenge for two weeks. LRE suppressed inflammatory cells and Th2 cytokines (IL-4, IL-5 and IL-13) in BALF and reduced IgE level in the serum. LRE also attenuated eosinophils infiltration and goblet cell hyperplasia in the lung tissues; as well as ameliorated airway remodelling by reducing smooth muscle thickness and reducing the expressions of TGF-β1 and Activin A positive cell in the lung tissues. LRE attenuated airway inflammation and remodelling in the prolonged allergen challenge of allergic asthma model. These findings suggest the therapeutic potential of LRE as an alternative for the management of allergic asthma.

Topics: Airway Remodeling; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Transforming Growth Factor beta1

Angiotensin (Ang)-(1-7) can reduce airway inflammation and airway remodeling in allergic asthma. Autophagy-related 5 (ATG5) has attracted wide attentions in asthma. However, the effects of Ang-(1-7) on ATG5-mediated autophagy in allergic asthma are unclear.. In this study, human bronchial epithelial cell (BEAS-2B) and human bronchial smooth muscle cell (HBSMC) were treated with different dose of Ang-(1-7) to observe changes of cell viability. Changes of ATG5 protein expression were measured in 10 ng/mL of interleukin (IL)-13-treated cells. Transfection of ATG5 small interference RNA (siRNA) or ATG5 cDNA in cells was used to analyze the effects of ATG5 on secretion of cytokines in the IL-13-treated cells. The effects of Ang-(1-7) were compared to the effects of ATG5 siRNA transfection or ATG5 cDNA transfection in the IL-13-treated cells. In wild-type (WT) mice and ATG5 knockout (ATG5. The results showed that ATG5 protein level was decreased with Ang-(1-7) dose administration in the IL-13-treated BEAS-2B and IL13-treated HBSMC. Ang-(1-7) played similar results to ATG5 siRNA that it suppressed the secretion of IL-25 and IL-13 in the IL-13-treated BEAS-2B cells, and inhibited the expression of transforming growth factor (TGF)-β1 and α-smooth muscle actin (α-SMA) protein in the IL-13-treated HBSMC cells. ATG5 cDNA treatment significantly increased the secretion of IL-25 and IL-13 and expression of TGF-β1 and α-SMA protein in IL-13-treated cells. Ang-(1-7) treatment suppressed the effects of ATG5 cDNA in the IL-13-treated cells. In OVA-induced WT mice, Ang-(1-7) treatment suppressed airway inflammation, remodeling and autophagy. ATG5 knockout also suppressed the airway inflammation, remodeling and autophagy.. Ang-(1-7) treatment suppressed airway inflammation and remodeling in allergic asthma through inhibiting ATG5, providing an underlying mechanism of Ang-(1-7) for allergic asthma treatment.

Topics: Airway Remodeling; Animals; Asthma; Autophagy-Related Protein 5; Disease Models, Animal; DNA, Complementary; Fibrosis; Humans; Inflammation; Interleukin-13; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Small Interfering; Transforming Growth Factor beta1

Airway remodeling is associated with severity and treatment insensitivity in asthma. This study aimed to investigate the effects of G protein-coupled receptor 120 (GPR120) stimulation on alleviating allergic inflammation and remodeling of airway epithelium.. Ovalbumin (OVA)-challenged BALB/c mice and type-2-cytokine (IL-4 and IL-13)-exposed 16HBE human bronchial epithelial cells were treated with GSK137647A, a selective GPR120 agonist. Markers of allergic inflammation and airway remodeling were determined.. GSK137647A attenuated inflammation and mucus secretion in airway epithelium of OVA-challenged mice. Stimulation of GPR120 in 16HBE suppressed expression of asthma-associated cytokines and cytokine-induced expression of pathogenic mucin-MUC5AC. These effects were abolished by co-treatment with AH7614, a GPR120 antagonist. Moreover, GPR120 stimulation in 16HBE cells reduced expression of fibrotic markers including fibronectin protein and ACTA2 mRNA and inhibited epithelial barrier leakage induced by type-2 inflammation via rescuing expression of zonula occludens-1 protein. Furthermore, GPR120 stimulation prevented the cytokine-induced airway epithelial remodeling via suppression of STAT6 and Akt phosphorylation.. Our findings suggest that GPR120 activation alleviates allergic inflammation and remodeling of airway epithelium partly through inhibition of STAT6 and Akt. GPR120 may represent a novel therapeutic target for diseases associated with remodeling of airway epithelium, including asthma.

Topics: Airway Remodeling; Animals; Asthma; Cytokines; Disease Models, Animal; Humans; Inflammation; Interleukin-13; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Proto-Oncogene Proteins c-akt; Receptors, G-Protein-Coupled; Signal Transduction; STAT6 Transcription Factor

Bronchoconstriction, along with inflammation and hyperresponsiveness is the characteristic feature associated with asthma, contributing to variable airflow obstruction, which manifests shortness of breath, cough and wheeze, etc. Histone deacetylases 8 (HDAC8) is the member of class I HDAC family and known to regulate microtubule integrity and muscle contraction. Therefore, we aimed to investigate the effects of HDAC8 inhibition in murine model of asthma using Pan-HDAC inhibitor curcumin (CUR) and HDAC8-specific inhibitor PCI-34051 (PCI), alone and in combination.. To develop asthmatic mouse model, Balb/c mice were sensitized and challenged with ovalbumin (OVA). CUR (10 mg/kg, pre, post, alone and combined treatment) and PCI (0.5 mg/kg), were administered through intranasal (i.n) route, an hour before OVA aerosol challenge. Effects of HDAC8 inhibition by CUR and PCI pretreatments were evaluated in terms of inflammation, oxidative stress and fibrosis markers. Efficacy of curcumin post-treatment (CUR(p)) was also evaluated simultaneously.. Inflammatory cell recruitment, oxidative stress (reactive oxygen species, nitric oxide), histamine and Immunoglobulin E (IgE) levels and expression of fibrosis markers including hydroxyproline, matrix metalloproteinases-9 and alpha smooth muscle actin (MMP-9 and α-SMA) were significantly reduced by CUR, CUR(p), PCI-alone and combined treatments. Protein expressions of HDAC8, Nuclear factor-κB (NF-κB) accompanied by MAPKs (mitogen-activated protein kinases) were significantly reduced by the treatments. Structural alterations were examined by histopathological analysis and linked with the fibrotic changes.. Present study indicates protective effects of HDAC8 inhibition in asthma using HDAC8 using CUR and PCI alone or in combination, attenuates airway inflammation, fibrosis and remodeling; hence, bronchoconstriction was accompanied through modulation of MAP kinase pathway.

Topics: Animals; Asthma; Curcumin; Disease Models, Animal; Fibrosis; Inflammation; Lung; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; Ovalbumin

To investigate the effects of corilagin on inflammation and collagen deposition in ovalbumin (OVA)-induced asthma mouse model and uncover the mechanism.. We constructed a mouse model of OVA-induced asthma. Enzyme-linked-immunosorbent serologic assays were conducted to detect the effects of corilagin on cytokines and Immunoglobulin E (IgE) production. Hematoxylin and eosin staining was used to show pathological features in lung tissues. Masson trichrome assay was used to examine collagen deposition. In addition, the lung function was detected by mouse lung function apparatus. Immunoblot was used to confirm the mechanism.. Corilagin alleviates OVA-induced cytokine and IgE production. In addition, corilagin alleviates OVA-induced pathological changes and collagen deposition in lung tissues. Corilagin also suppressed airway resistance and lung function in mice. Mechanically, corilagin activated the adenosine monophosphate-activated protein kinase (AMPK) pathway in lung tissues.. Corilagin attenuates airway inflammation and collagen deposition in OVA-induced asthmatic mice via AMPK pathway.

Topics: AMP-Activated Protein Kinases; Animals; Asthma; Bronchoalveolar Lavage Fluid; Collagen; Cytokines; Disease Models, Animal; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Asthma, a chronic airway inflammatory disease, has a complicated pathogenesis and limited therapeutic treatment. Evidence shows that the intestinal microbiota exhibits crucial functional interaction with asthma syndrome. Liubao tea (LBT), a type of postfermented tea in China, positively modulates gut microbiota. However, the potential benefits of LBT extract (LBTE) for allergic asthma are still not understood. Herein, the anti-inflammatory effects of LBTE and its modulation of the gut microbiota of asthmatic mice induced by ovalbumin were explored. The results demonstrate that LBTE significantly inhibited airway hyper-responsiveness and restrained the proliferation of proinflammatory cytokines and inflammatory cells associated with allergic asthma. Additionally, LBTE suppressed inflammatory infiltration, mucus secretion, and excessive goblet cell production by downregulating the gene expression of inflammatory indicators. Interestingly, fecal microbiota transplantation results further implied that the modulation of LBTE on gut microbiota played an essential role in alleviating airway inflammatory symptoms of allergic asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Gastrointestinal Microbiome; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Tea

Interleukin-17A (IL-17A) levels are elevated in patients with asthma. Ferroptosis has been identified as the non-apoptotic cell death type associated with asthma. Data regarding the relation of ferroptosis with asthma and the effect of IL-17A on modulating ferroptosis in asthma remain largely unclear. The present work focused on investigating the role of IL-17A in allergic asthma-related ferroptosis and its associated molecular mechanisms using public datasets, clinical samples, human bronchial epithelial cells, and an allergic asthma mouse model. We found that IL-17A was significantly upregulated within serum in asthma cases. Adding IL-17A significantly increased ferroptosis within human bronchial epithelial cells (BEAS-2B). In ovalbumin (OVA)-induced allergic asthmatic mice, IL-17A regulated and activated lipid peroxidation induced ferroptosis, whereas IL-17A knockdown effectively inhibited ferroptosis in vivo by protection of airway epithelial cells via the xCT-GSH-GPX4 antioxidant system and reduced airway inflammation. Mouse mRNA sequencing results indicated that the tumor necrosis factor (TNF) pathway was the differential KEGG pathway in the OVA group compared to healthy controls and the OVA group compared to the IL-17A knockout OVA group. We further used N-acetylcysteine (TNF inhibitor) to inhibit the TNF signaling pathway, which was found to protect BEAS-2B cells from IL-17A induced lipid peroxidation and ferroptosis damage. Our findings reveal a novel mechanism for the suppression of ferroptosis in airway epithelial cells, which may represent a new strategy for the use of IL-17A inhibitors against allergic asthma.

Topics: Animals; Asthma; Disease Models, Animal; Epithelial Cells; Ferroptosis; Humans; Inflammation; Interleukin-17; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Gekko gecko is used as a traditional medicine for various diseases including respiratory disorders in northeast Asian countries, mainly Korea, Japan, and China.. Allergic asthma is a chronic respiratory disease caused by an inappropriate immune response. Due to the recent spread of coronavirus disease 2019, interest in the treatment of pulmonary disorders has rapidly increased. In this study, we investigated the anti-asthmatic effects of G. gecko extract (GGE) using an established mouse model of ovalbumin-induced asthma.. To evaluate the anti-asthmatic effects of GGE, we evaluated histological changes and the responses of inflammatory mediators related to allergic airway inflammation. Furthermore, we investigated the regulatory effects of GGE on type 2 helper T (Th2) cell activation.. Administration of GGE attenuated asthmatic phenotypes, including inflammatory cell infiltration, mucus production, and expression of Th2 cytokines. Furthermore, GGE treatment reduced Th2 cell activation and differentiation.. These results indicate that GGE alleviates allergic airway inflammation by regulating Th2 cell activation and differentiation.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; COVID-19; Cytokines; Female; Flow Cytometry; Immunoglobulin E; Inflammation Mediators; Lung; Medicine, East Asian Traditional; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Pandemics; Plant Extracts; Th2 Cells; Tryptamines

Microbial interventions against allergic asthma have robust epidemiologic underpinnings and the potential to recalibrate disease-inducing immune responses. Oral administration of OM-85, a standardized lysate of human airways bacteria, is widely used empirically to prevent respiratory infections and a clinical trial is testing its ability to prevent asthma in high-risk children. We previously showed that intranasal administration of microbial products from farm environments abrogates experimental allergic asthma.. We sought to investigate whether direct administration of OM-85 to the airway compartment protects against experimental allergic asthma; and to identify protective cellular and molecular mechanisms activated through this natural route.. Different strains of mice sensitized and challenged with ovalbumin or Alternaria received OM-85 intranasally, and cardinal cellular and molecular asthma phenotypes were measured. Airway transfer experiments assessed whether OM-85-treated dendritic cells protect allergen-sensitized, OM-85-naive mice against asthma.. Airway OM-85 administration suppressed allergic asthma in all models acting on multiple innate and adaptive immune targets: the airway epithelium/IL-33/ILC2 axis, lung allergen-induced type 2 responses, and dendritic cells whose Myd88/Trif-dependent tolerogenic reprogramming was sufficient to transfer OM-85-induced asthma protection.. We provide the first demonstration that administering a standardized bacterial lysate to the airway compartment protects from experimental allergic asthma by engaging multiple immune pathways. Because protection required a cumulative dose 27- to 46-fold lower than the one reportedly active through the oral route, the efficacy of intranasal OM-85 administration may reflect its direct access to the airway mucosal networks controlling the initiation and development of allergic asthma.

Topics: Allergens; Animals; Asthma; Cell Extracts; Dendritic Cells; Disease Models, Animal; Epithelium; Humans; Immunity, Innate; Interleukin-33; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin

The administration of L-glutamine (Gln) suppresses allergic airway inflammation via the rapid upregulation of MAPK phosphatase (MKP)-1, which functions as a negative regulator of inflammation by deactivating p38 and JNK mitogen-activated protein kinases (MAPKs). However, the role of endogenous Gln remains to be elucidated. Therefore, we investigated the mechanism by which endogenous Gln regulates MKP-1 induction and allergic airway inflammation in an ovalbumin-based murine asthma model.. We depleted endogenous Gln levels using L-γ-glutamyl-p-nitroanilide (GPNA), an inhibitor of the Gln transporter ASCT2 and glutamine synthetase small interfering siRNA. Lentivirus expressing MKP-1 was injected to achieve overexpression of MKP-1. Asthmatic phenotypes were assessed using our previously developed ovalbumin-based murine model, which is suitable for examining sequential asthmatic events, including neutrophil infiltration. Gln levels were analyzed using a Gln assay kit.. GPNA or glutamine synthetase siRNA successfully depleted endogenous Gln levels. Importantly, homeostatic MKP-1 induction did not occur at all, which resulted in prolonged p38 MAPK and cytosolic phospholipase A. Gln deficiency leads to the impairment of MKP-1 induction and activation of p38 MAPK and cPLA

Topics: Animals; Asthma; Dual Specificity Phosphatase 1; Glutamate-Ammonia Ligase; Glutamine; Humans; Inflammation; Lung; Mice; Ovalbumin; p38 Mitogen-Activated Protein Kinases; RNA, Small Interfering

Sphagneticola trilobata (L.) Pruski is used in traditional medicine in Brazil for inflammatory diseases treatment including asthma. The diterpene kaurenoic acid (KA) is one of its active compounds, but whether KA activity could explain the traditional use of S. trilobata in asthma is unknown.. Investigate KA effect and mechanisms in asthma.. Experimental asthma was induced by ovalbumin immunization and challenge in male Swiss mice. KA (0.1-10 mg/kg, gavage) was administered 1 h before the ovalbumin challenge. Total leukocytes, eosinophil, and mast cell were counted in bronchoalveolar lavage fluid (BALF), and lung histopathology was performed. Lung mRNA expression of Th2 and regulatory T cells markers, and BALF type 2 cytokine production were quantitated. NFκB activation and oxidative stress-related components in pulmonary tissue were measured.. KA inhibited the migration of total leukocytes and eosinophils to BALF, reduced lung histopathology (inflammatory cells and mast cells), mRNA expression of IL-33/ST2, STAT6/GATA-3 and NFκB activation in the lung, and reduced IL-33, IL-4, IL-5 production in the BALF. KA also reduced the mRNA expression of iNOS and gp91. KA prevents antigen-induced asthma by down-regulating Th2 and NFκB/cytokine-related pathways, and up-regulating Nrf2 and regulatory T cells' markers. Thus, explaining the ethnopharmacological use of S. trilobata for the treatment of lung inflammatory diseases.

Topics: Animals; Asteraceae; Asthma; Cytokines; Disease Models, Animal; Diterpenes; Dose-Response Relationship, Drug; GATA3 Transcription Factor; Male; Mice; NF-E2-Related Factor 2; NF-kappa B; Ovalbumin; STAT6 Transcription Factor; Th2 Cells

Asthma being an inflammatory disease of the airways lead to structural alterations in lungs which often results in the severity of the disease. Curcumin, diferuloylmethane, is well known for its medicinal properties but its anti-inflammatory potential via Histone deacetylase inhibition (HDACi) has not been revealed yet. Therefore, we have explored here, anti-inflammatory and anti-fibrotic potential of intranasal curcumin via HDAC inhibition and compared its potential with Sodium butyrate (SoB), a known histone deacetylase inhibitor of Class I and II series. Anti-inflammatory potential of SoB, has been investigated in cancer but not been studied in asthma before.. In present study, ovalbumin (OVA) was used to sensitize Balb/c mice and later exposed to (1%) OVA aerosol. Curcumin (5 mg/kg) and Sodium butyrate (50 mg/kg) was administered through intranasal route an hour before OVA aerosol challenge. Efficacies of SoB and Curcumin as HDAC inhibitors were evaluated in terms of different inflammatory parameters like, total inflammatory cell count, reactive oxygen species (ROS), histamine release, nitric oxide and serum IgE levels. Inflammatory cell recruitment was analyzed by H&E staining and structural alterations were revealed by Masson's Trichrome staining of lung sections.. Enhanced Matrix Metalloproteinase-2 and 9 (MMP-2 and MMP-9) activities were observed in bronchoalveolar lavage fluid (BALF) of asthmatic mice by gelatin zymography which was inhibited in both treatment groups. Protein expressions of MMP-9, HDAC 1, H3acK9 and NF-kB p65 were modulated in intranasal curcumin and SoB pretreatment groups.. This is the first report where intranasal curcumin inhibited asthma severity via affecting HDAC 1 (H3acK9) leading to NF-kB suppression in mouse model of allergic asthma.

Topics: Administration, Intranasal; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Butyric Acid; Curcumin; Disease Models, Animal; Fibrosis; Histone Deacetylase Inhibitors; Immunoglobulin E; Inflammation; Lung; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; Ovalbumin

Previous studies have shown Interleukin (IL)-17A as an important contributor to the development of severe asthma, which is mainly characterized by neutrophilic inflammation and less response to corticosteroids. Consequently, the IL-17A-neutrophil axis could be a potential therapeutic target. Previously, we constructed a recombinant. DO11.10 mice were divided into four groups: phosphate buffered saline (PBS), asthma, rMS and MS. This murine model of neutrophilic asthma was established with ovalbumin (OVA) challenge, whereby PBS, rMS and MS were administered intranasally. Anti-inflammatory effects on inflammatory cell infiltration and expression of inflammatory mediators in bronchoalveolar lavage fluid (BALF) were evaluated, along with histopathological changes in lung tissues.. A sustained high-titer IL-17A autoantibody was detected in sera of the rMS group. Compared to the asthma group, the number of neutrophils, IL-17A, CXCL-1 levels and MPO activity in the rMS group were all significantly reduced (. rMS ameliorated airway inflammation in mice with neutrophilic asthma caused by inducing IL-17A autoantibody and regulating the IL-17A-neutrophil axis, thus offering a possible novel treatment for neutrophilic asthma.

Topics: Adrenal Cortex Hormones; Animals; Anti-Inflammatory Agents; Asthma; Disease Models, Animal; Inflammation; Inflammation Mediators; Interleukin-17; Mice; Mice, Inbred BALB C; Mycobacterium smegmatis; Ovalbumin; Phosphates

Childhood asthma is the most universal chronic disease, with significant cases reported. Despite the current progress in treatment, prognosis remains poor and the existing drugs cause serious side effects. This investigation explored the mechanisms and use of miR-335-5p on childhood asthma therapy. MiR-335-5p and ATG5 expression was analyzed in clinical plasma samples through RT-qPCR. Airway smooth muscle cells (ASMCs) were cultured, and transfected with miR-335-5p mimic, miR-335-5p inhibitor, and pcDNA3.1-ATG5, or co-transfected with miR-335-5p mimic + pcDNA3.1-ATG5. Asthma cell models were constructed through TGF-β1, and animal models through ovalbumin (OVA). Monocyte-macrophage infiltration in bronchoalveolar lavage fluid (BALF) was determined by May-Grunwald-Giemsa staining, and collagen in lung tissue was assessed via Masson staining. Relationship between miR-335-5p and ATG5 was detected by dual-luciferase assay. Cell proliferation was detected by MTT assay. MiR-335-5p and ATG5 RNA expression was determined by RT-qPCR. Collagen I, collagen III, α-SMA, ATG5, LC3I/II, Beclin-1, and p62 protein expression levels in ASMCs were detected by western blot. MiR-335-5p expression was low, but ATG5 expression was high in childhood asthma. Versus OVA+ mimic NC group, the number of eosinophil and collagen in OVA+ miR-335-5p mimic group were reduced. In contrast to TGF-β1 + mimic NC group, TGF-β1 + miR-335-5p mimic group reduced inflammatory, airway fibrosis, and autophagy in ASMCs. ATG5 was miR-335-5p target. Overexpressing ATG5 significantly reversed the inhibitory effects of miR-335-5p on inflammatory response, fibrosis, and autophagy in ASMCs. Overall, the study concludes that MiR-335-5p alleviate inflammatory response, airway fibrosis, and autophagy in childhood asthma through targeted regulation of ATG5.

Topics: Animals; Asthma; Autophagy; Autophagy-Related Protein 5; Bronchoalveolar Lavage Fluid; Cell Proliferation; Cells, Cultured; Child; Disease Models, Animal; Female; Humans; Male; MicroRNAs; Myocytes, Smooth Muscle; Ovalbumin; Respiratory System; Signal Transduction; Transforming Growth Factor beta

Asthma is a chronic disease closely related to airway inflammation. It has been proven that type 2 innate lymphoid cells (ILC2s) play an essential role in airway inflammation in asthma. Furthermore, there is growing evidence that Follistatin-like 1 (FSTL1) can participate in various inflammatory reactions mediated by the JAK/STAT signaling pathway, among others. Therefore, we put forward a new hypothesis: FSTL1 promotes asthmatic airway inflammation by activating ILC2. This study generated an ovalbumin-sensitized asthma model in C57BL/6 and Fstl1

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Follistatin; Immunity, Innate; Inflammation; Lung; Lymphocytes; Mice; Mice, Inbred C57BL; Ovalbumin

Phlomis umbrosa Turczaninow has been used as a tradition herbal medicine for treating various inflammatory diseases.. In present study, we explored the effects of P. umbrosa on asthma induced by ovalbumin (OVA) and elucidated the mechanism via in vivo verification and network pharmacology prediction.. The animals were intraperitoneally injected OVA on day 1 and 14, followed by OVA inhalation on days 21, 22, and 23. The animals were daily treated P. umbrosa extract (PUE, 20 and 40 mg/kg) by oral gavage from day 18 to day 23.. PUE significantly decreased airway hyperresponsiveness, eosinophilia, and the production of inflammatory cytokines and OVA specific immunoglobulin E in animals with asthma, along with a reduction in airway inflammation and mucus secretion in lung tissue. In network analysis, antiasthmatic effects of PUE were closely related with suppression of mitogen-activated protein kinases and matrix metalloproteinases (MMPs). Consistent with the results from network analysis, PUE suppressed the phosphorylation of ERK and p65, which was accompanied by a decline in MMP-9 expression.. Administration of PUE effectively reduced allergic responses in asthmatic mice, which was associated with the suppressed phosphorylation of ERK and p65, and expression of MMP-9. These results indicate that PUE has therapeutic potential to treat allergic asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Disease Models, Animal; Dose-Response Relationship, Drug; Extracellular Signal-Regulated MAP Kinases; Female; Inflammation; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; Network Pharmacology; Ovalbumin; Phlomis; Phosphorylation; Plant Extracts; Respiratory Hypersensitivity; Transcription Factor RelA

Jia-Wei Bu-Shen-Yi-Qi formula (JWBSYQF), a Chinese herbal formula, is a commonly used prescription for treating asthma patients. However, the targeted proteins associated with JWBSYQF treatment remain unknown.. Present study aims to evaluate the therapeutic efficacy of JWBSYQF and identify the targeted proteins in addition to functional pathways.. The ovalbumin (OVA)-induced murine asthma model was established to explore the therapeutic effect of JWBSYQF treatment. Proteomic profiling and quantifications were performed using data-independent acquisition (DIA) methods. Differentially expressed proteins (DEPs) were validated via western blot (WB) and immunohistochemistry (IHC).. A murine asthma model was made by OVA sensitization and challenge, and JWBSYQF (2.25, 4.50, 9,00 g/kg body weight) or dexamethasone (1 mg/ kg body weight) were administered orally. Airway hyperresponsiveness (AHR) to methacholine (Mch), inflammatory cell counts and classification in bronchoalveolar lavage fluid (BALF), lung histopathology, and cytokine levels were measured. Furthermore, DIA proteomic analyses were performed to explore the DEPs targeted by JWBSYQF and were further validated by WB and IHC.. Our results exhibited that JWBSYQF attenuated AHR which was mirrored by decreased airway resistance and increased lung compliance. In addition, JWBSYQF-treated mice showed reduced inflammatory score, mucus hypersecretion, as well as reduced the number of BALF leukocytes along with decreased content of BALF Th2 inflammatory cytokines (IL-4, IL-5, IL-13) and serum IgE. Proteomics analysis identified 704 DEPs between the asthmatic mice and control group (MOD vs CON), and 120 DEPs between the JWBSYQF-treatment and the asthmatic mice (JWB-M vs MOD). A total of 33 overlapped DEPs were identified among the three groups. Pathway enrichment analysis showed that DEPs were significantly enriched in IL-17 signaling pathway, in which DEPs, Lcn2, TGF-β1, Gngt2, and Ppp2r5e were common DEPs between three experimental groups. WB and IHC results further validated expressional levels and tendency of these proteins. Our results also showed that JWBSYQF affects mitogen activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) signaling pathways, that are activated by IL-17 signaling.. The present study suggested that JWBSYQF could attenuate AHR and airway inflammation in OVA-induced asthmatic mice. In addition, proteomics analysis revealed that suppression of IL-17 signaling pathways contributes to the therapeutic effects of JWBSYQF.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Interleukin-17; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Proteomics; Signal Transduction

Asthma characterized by airway remodeling is a multiple pulmonary disease, which is associated with various physiological processes including inflammation reaction, immune response, oxidative stress and autophagy.. This study aimed to investigate whether these processes are modulated by the total glucosides of Paeonia lactiflora Pall (TGP), and its active compound paeoniflorin (PF) with anti-inflammatory and immune-regulatory effects could alleviate ovalbumin (OVA)-induced mouse asthma.. In vivo, models of mouse asthma were established by intraperitoneally with a mixture of OVA and aluminum hydroxide, plus a single nasal injected with OVA to female C57BL/6 mice. The results were observed with PET imaging, TEM, RT-PCR, western blotting. In vitro, CD4. TGP, either in its crude or processed form, and PF effectively ameliorated lung injury in mice induced by OVA, regulated immune/inflammatory response by inhibiting the release of pro-inflammatory cytokines, thereby decreasing Th2 cell proportion, inhibited oxidative stress by recovering mitochondrial membrane potential and regulating metabolic activity in dose-dependent manner. Moreover, PF could inhibit autophagy by regulating mitochondrial function. In addition, the therapeutic effects of TGP and PF on pulmonary injury in asthmatic mice were not affected by processing.. PF may be a valuable agent in ameliorating inflammation and immune response in asthmatic mice, and the possible mechanism involved in this response rang may from oxidative stress to autophagy.

Topics: Animals; Asthma; Autophagy; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Glucosides; Immunity; Inflammation; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Monoterpenes; Ovalbumin; Oxidative Stress

Maternal gestational exposures to traffic and urban air pollutant particulates have been linked to increased risk and/or worsening asthma in children; however, mechanisms underlying this vertical transmission are not entirely understood. It was postulated that gestational particle exposure might affect the ability to elicit specialized proresolving mediator (SPM) responses upon allergen encounter in neonates. Lipidomic profiling of 50 SPMs was performed in lungs of neonates born to mice exposed to concentrated urban air particles (CAP), diesel exhaust particles (DEP), or less immunotoxic titanium dioxide particles (TiO2). While asthma-like phenotypes were induced with identical eosinophilia intensity across neonates of all particle-exposed mothers, levels of LXA4, HEPE and HETE isoforms, and HDoHe were only decreased by CAP and DEP only but not by TiO2. However, RvE2 and RvD1 were inhibited by all particles. In contrast, isomers of Maresin1 and Protectin D1 were variably elevated by CAP and DEP, whereas Protectin DX, PGE2, and TxB2 were increased in all groups. Only Protectin D1/DX, MaR1(n-3,DPA), 5(S),15(S)-DiHETE, PGE2, and RvE3 correlated with eosinophilia but the majority of other analytes, elevated or inhibited, showed no marked correlation with inflammation intensity. Evidence indicates that gestational particle exposure leads to both particle-specific and nonspecific effects on the SPM network.

Topics: Allergens; Animals; Animals, Newborn; Asthma; Disease Susceptibility; Eosinophilia; Female; Inflammation Mediators; Inhalation Exposure; Lung; Maternal Exposure; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Titanium; Vehicle Emissions

Airway remodelling is a critical feature of chronic lung diseases. Epithelial-mesenchymal transition (EMT) represents an important source of myofibroblasts, contributing to airway remodelling. Here, we investigated the sphingosine-1-phosphate (S1P) role in EMT and its involvement in asthma-related airway dysfunction.. A549 cells were used to assess the S1P effect on EMT and its interaction with TGF-β signalling. To assess the S1P role in vivo and its impact on lung function, two experimental models of asthma were used by exposing BALB/c mice to subcutaneous administration of either S1P or ovalbumin (OVA).. Following incubation with TGF-β or S1P, A549 acquire a fibroblast-like morphology associated with an increase of mesenchymal markers and down-regulation of the epithelial. These effects are reversed by treatment with the TGF-β receptor antagonist LY2109761. Systemic administration of S1P to BALB/c mice induces asthma-like disease characterized by mucous cell metaplasia and increased levels of TGF-β, IL-33 and FGF-2 within the lung. The bronchi harvested from S1P-treated mice display bronchial hyperresponsiveness associated with overexpression of the mesenchymal and fibrosis markers and reduction of the epithelial.The S1P-induced switch from the epithelial toward the mesenchymal pattern correlates to a significant increase of lung resistance and fibroblast activation. TGF-β blockade, in S1P-treated mice, abrogates these effects. Finally, inhibition of sphingosine kinases by SK1-II in OVA-sensitized mice, abrogates EMT, pulmonary TGF-β up-regulation, fibroblasts recruitment and airway hyperresponsiveness.. Targeting S1P/TGF-β axis may hold promise as a feasible therapeutic target to control airway dysfunction in asthma.

Topics: Airway Remodeling; Animals; Asthma; Epithelial Cells; Epithelial-Mesenchymal Transition; Lysophospholipids; Mice; Mice, Inbred BALB C; Ovalbumin; Sphingosine; Transforming Growth Factor beta; Transforming Growth Factor beta1

Salidroside (Sal) a bioactive component extracted from Rhodiola rosea is remarkable for its anti-asthmatic effects. The study aimed to explore the molecular mechanism of Sal in airway inflammation and remodeling in asthmatic mice and provide a novel theoretical basis for asthma treatment.. An asthmatic mouse model was established via ovalbumin (OVA) treatment, followed by injection of Sal and transfection of miR-323-3p-mimic and sh- suppressor of cytokine signaling 5 (SOCS5). Expressions of miR-323-3p, SOCS5 mRNA, collagen (COL)-I, and COL-III were detected via reverse transcription quantitative polymerase chain reaction. SOCS5 protein level was detected via Western blot. Levels of IgE, IL-13, IL-4, and IL-5 were detected via enzyme-linked immunosorbent assay. Inflammatory cell infiltration was observed via hematoxylin-eosin staining. Collagen disposition was observed via Masson staining. Resistance index (RI) of airway hyperresponsiveness, and the number of total cells, inflammatory cells (eosinophil, macrophage, neutrophil, and lymphocyte) in bronchoalveolar lavage fluid (BALF) were observed. The binding relationship between miR-323-3p and SOCS5 was predicted through the RNA22 website and verified via dual-luciferase reporter assay.. miR-323-3p was highly expressed in OVA-treated mice. Sal treatment reduced inflammatory cell infiltration, COL disposition, miR-323-3p expression, and IgE, IL-13, IL-4, IL-5, COL-I, and COL-III levels, RI value, and the number of total cells and inflammatory cells in BALF. miR-323-3p inhibited SOCS5 transcription. miR-323-3p overexpression or SOCS5 downregulation reversed the protecting role of Sal in asthmatic mice.. Sal inhibited miR-323-3p expression to promote SOCS5 transcription, thereby attenuating airway inflammation and remodeling in asthmatic mice.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Glucosides; Inflammation; Lung; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Phenols; Signal Transduction; Suppressor of Cytokine Signaling Proteins

RNA-binding protein HuR (

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Allergens; Animals; Anti-Inflammatory Agents; Asthma; Benzopyrans; Cells, Cultured; Disease Models, Animal; ELAV-Like Protein 1; Humans; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Middle Aged; Ovalbumin; Pyrrolidines; Th2 Cells; Young Adult

Although exposure to ambient particulate matter (PM) is linked to asthma, the health effects of co-existing vapor-phase organic pollutants (vapor) and their combined effects with PM on this disease are poorly understood. We used a murine asthma model to test the hypothesis that exposure to vapor would enhance allergic sensitization and this effect would be further strengthened by co-existing PM. We found that vapor and PM each individually exerted adjuvant effects on OVA sensitization. Co-exposure to vapor and PM during sensitization further enhanced allergic lung inflammation and OVA-specific antibody production which was accompanied by pulmonary cytokine/chemokine milieu that favored T-helper 2 immunity (i.e. increased IL-4, downregulation of Il12a and Ifng, and upregulation of Ccl11 and Ccl8). TNFα, IL-6, Ccl12, Cxcl1 and detoxification/antioxidant enzyme responses in the lung were pollutant-dependent. Inhibition of lipopolysaccharide-induced IL-12 secretion from primary antigen-presenting dendritic cells correlated positively with vapor's oxidant potential. In conclusion, concurrent exposure to vapor and PM led to significantly exaggerated adjuvant effects on allergic lung inflammation which were more potent than that of each pollutant type alone. These findings suggest that the effects of multi-component air pollution on asthma may be significantly underestimated if research only focuses on a single air pollutant (e.g., PM).

Topics: Animals; Asthma; Cytokines; Down-Regulation; Drug Interactions; Female; Gene Expression Regulation; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Particle Size; Particulate Matter; RNA, Messenger; Th2 Cells; Up-Regulation; Volatile Organic Compounds

Although lung macrophages are directly exposed to external stimuli, their exact immunologic roles in asthma are still largely unknown. The aim of this study was to investigate the anti-asthmatic effect of Acinetobacter lwoffii in terms of lung macrophage modulation.. Six-week-old female BALB/c mice were sensitized and challenged with ovalbumin (OVA) with or without intranasal administration of A. lwoffii during the sensitization period. Airway hyperresponsiveness and inflammation were evaluated. Using flow cytometry, macrophages were subclassified according to their activation status. In the in vitro study, a murine alveolar macrophage cell line (MH-S) treated with or without A. lwoffii before IL-13 stimulation were analysed by quantitative RT-PCR.. In a murine asthma model, the number of inflammatory cells, including macrophages and eosinophils, decreased in mice treated with A. lwoffii (A. lwoffii/OVA group) compared with untreated mice (OVA group). The enhanced expression of MHCII in macrophages in the OVA group was decreased by A. lwoffii treatment. M2 macrophage subtypes were significantly altered. A. lwoffii treatment decreased CD11b. Intranasal A. lwoffii exposure suppresses asthma development by suppressing the type 2 response via modulating lung macrophage activation, shifting M2a and M2c macrophages to M2b macrophages.

Topics: Acinetobacter; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Humans; Immunity, Innate; Inflammation; Lung; Lymphocytes; Macrophage Activation; Mice; Mice, Inbred BALB C; Ovalbumin

Bronchial asthma (BA) is one of the most common chronic inflammatory disease of airways. There are huge experimental data indicating that Th2-cytokines IL-4 and IL-13 play a key role in BA pathogenesis. They are implicated in the IgE synthesis, eosinophil infiltration to the lungs and in the development of airway hyperreactivity (AHR), that makes these cytokines the promising targets. Neutralization of IL-4 and IL-13 or its common receptor chain (IL-4Rα) by monoclonal antibodies substantially reduce asthma symptoms. RNA interference provides a novel method for regulation of gene expression by siRNA molecules. In this study we evaluated whether the siRNA targeted to IL-4 and IL-13 reduce BA symptoms in mice model. Experimental BA was induced in BALB/c mice by sensitization to ovalbumin allergen followed by intranasal challenge. The intranasal delivery of siRNAs targeted to IL-4 and IL-13 inhibited the lung expression of these cytokines by more than 50% that led to the attenuation of AHR and pulmonary inflammation; the quantity of eosinophils in lungs which are one of the major inflammatory cells involved in allergic asthma pathogenesis decreased by more than 50% after siRNA treatment. These data support the possibility of a dual IL-4 and IL-13 inhibition by locally delivered siRNAs which in turn leads to the suppression of allergen-induced pulmonary inflammation and AHR.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Eosinophils; Immunoglobulin E; Inflammation; Interleukin-13; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Small Interfering

Gerberae Piloselloidis Herba is widely used to treat cough and asthma in China. However, its effects on allergic asthma as related to its chemical compositions have not been fully elucidated, and there is a scarcity of methods to determine multi-component contents for quality control. In this study, protective effects of Gerberae Piloselloidis Herba on ovalbumin-induced asthma models were investigated, while qualitative and quantitative analyses of multiple constituents in Gerberae Piloselloidis Herba were conducted by using an ultrahigh-performance liquid chromatography-Q Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry and an ultrahigh-performance liquid chromatography-photodiode array detection. The results showed that Gerberae Piloselloidis Herba could significantly mitigate asthma symptoms, reduce eosinophils counts in the bronchoalveolar lavage fluid, as well as decrease IgE, IL-5, and IL-13 concentration, and inflammatory cellular infiltration in lung tissues. A total of 51 compounds were tentatively identified, in which the content of 10 representative compounds was determined in 24 batches of Gerberae Piloselloidis Herba by using an ultrahigh-performance liquid chromatography method with good linearity, precision, repeatability, accuracy, and stability. This research presents a comprehensive strategy combining biological activity evaluation with chemical profiling, providing a useful and comprehensive reference for further application and quality control of Gerberae Piloselloidis Herba.

Topics: Animals; Asthma; Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Mass Spectrometry; Mice; Ovalbumin

A certain population of asthma patients is resistant to steroid therapy, whereas the mechanisms remain unclear. One of characteristic features of steroid-resistant asthma patients is severe airway eosinophilia based on type-2 inflammation. Aims of this study were: 1) to develop a murine model of steroid-resistant asthma, 2) to elucidate that predominant cellular source of a type-2 cytokine, IL-5 was group 2 innate lymphoid cells (ILC2s), 3) to analyze pathogenic alteration of ILC2s in the severe asthma, and 4) to evaluate therapeutic potential of anti-IL-5 monoclonal antibody (mAb) on the steroid-resistant asthma. Ovalbumin (OVA)-sensitized BALB/c mice were intratracheally challenged with OVA at 5 or 500 μg/animal 4 times. Development of airway eosinophilia and remodeling in 5-μg OVA model were significantly suppressed by 1 mg/kg dexamethasone, whereas those in 500-μg OVA model were relatively insensitive to the dose of dexamethasone. ILC2s isolated from the lung of the steroid-insensitive model (500-μg OVA) produced significantly larger amounts of IL-5 in response to IL-33/TSLP than ILC2s from the steroid-sensitive model (5-μg OVA). Interestingly, TSLP receptor expression on ILC2s was up-regulated in the steroid-insensitive model. Treatment with anti-IL-5 mAb in combination with dexamethasone significantly suppressed the airway remodeling of the steroid-insensitive model. In conclusion, multiple intratracheal administration of a high dose of antigen induced steroid-insensitive asthma in sensitized mice. IL-5 was mainly produced from ILC2s, phenotype of which had been pathogenically altered probably through the up-regulation of TSLP receptors. IL-5 blockage could be a useful therapeutic strategy for steroid-resistant asthma.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Immunity, Innate; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Steroids

Topics: Administration, Oral; Animals; Asthma; Bronchoalveolar Lavage Fluid; Inflammation; Lactobacillus plantarum; Mice; Mice, Inbred BALB C; Ovalbumin

Asian sand dust (ASD), which mainly originates in China and Mongolia in the spring and blows into Korea, can exacerbate respiratory and immunological diseases. This study aims to observe effects of co-exposure to ASD on ovalbumin (OVA)-induced asthmatic lung inflammation and of treatment with a phosphodiesterase 7 (PDE7) inhibitor in a mouse model. The challenge with OVA increased airway hyperresponsiveness (AHR) and inflammatory cell infiltration into the lung tissue. Interleukin (IL)-13, tumor necrosis factor-alpha, monocyte-protein-1, mucin, and antigen-specific IgE and IgG1 production increased in mouse serum. The co-exposure of ASD significantly exacerbated these effects in this asthma model. Notably, the administration of a PDE7 inhibitor, BRL-50481 (BRL), significantly reduced AHR, infiltration of inflammatory cells into the lungs, and the levels of type 2 T helper cell-related cytokines, antigen-specific immunoglobulins, and mucin. Thus, the administration of BRL ameliorated OVA-induced allergic asthmatic responses exacerbated by co-exposure to ASD. This study suggests that PDE7 inhibition can be a therapeutic strategy for inflammatory lung diseases and asthma via the regulation of T lymphocytes and reduction of IL-13, and, consequently, mucin production.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cyclic Nucleotide Phosphodiesterases, Type 7; Cytokines; Disease Models, Animal; Dust; Fluorescent Antibody Technique; Inhalation Exposure; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Sand

The present study aims to investigate the effects of CCR3 gene knockout in bone marrow cells (CCR3-KO) on the mouse model of combined allergic rhinitis and asthma syndrome (CARAS). It was found that CCR3-KO significantly reduced eosinophil (EOS) migration into the nasal (NALF) and bronchoalveolar (BALF) cavities of mice, and decreased Th2 cytokines (such as, IL-4, IL-5 and IL-13) levels in nasal mucosa and lung tissues. In addition, histological analysis showed that the damage degree of nasal mucosa structure in ovalbumin (OVA) modulated CCR3-KO mice was significantly less than that in OVA modulated Wild type (WT) mice, with reduced inflammatory cell infiltration and nasal mucus secretion. The infiltration of inflammatory cells in lung tissue was significantly reduced, and the proliferation of lung smooth muscle layer and extracellular matrix (ECM) production were decreased. Symptom analysis showed that CCR3-KO can reduced allergic rhinitis (AR) signals as nose scratching and sneezing. It was also found CCR3-KO reduce OVA-induced weight loss. The results showed that CCR3-KO could reduce the symptoms of allergic inflammation in CARAS mice by reducing airway inflammatory cell infiltration and down-regulating the expression of Th2 cytokines, and CCR3 gene could be used as a target gene for the treatment of CARAS.

Topics: Allergens; Animals; Asthma; Bone Marrow Cells; Bronchoalveolar Lavage Fluid; Cytokines; Eosinophils; Immunoglobulin E; Lung; Mice, Inbred C57BL; Mice, Knockout; Nasal Lavage Fluid; Nasal Mucosa; Ovalbumin; Receptors, CCR3; Rhinitis, Allergic; Syndrome; Th2 Cells

The inflammasome NOD-like receptor (NLR) family, the pyrin domain containing 3 (NLRP3) is closely associated with exacerbation of asthma as endotoxin (lipopolysaccharide, LPS) is one of its activators present in the environment. Present study is undertaken to investigate anti-inflammatory effects of a well known phytochemical, curcumin, which might regulate LPS exposed asthma exacerbations by modulating NLRP3 activation if given through intranasal route. Balb/c mice were sensitized with intraperitoneal injection of OVA (Ovalbumin; 100 μg of OVA with alum) from day 1 to 8 and exposed to LPS with 1% OVA aerosol from day 9 to 15. LPS (0.1 μg) was given an hour before sensitization and OVA-aerosol challenge. Significant decrease in inflammatory cell recruitment and restoration of structural changes in lungs, alterations in mRNA and protein expressions of TLR-4, NF-κB, NLRP3, Caspase-1, IL-1β, MMP-9, IL-5 and IL-17 in intranasal curcumin alone and corticosteroid combined pretreatment group.

Topics: Administration, Intranasal; Animals; Asthma; Caspase 1; Curcumin; Dexamethasone; Inflammasomes; Interleukin-1beta; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Transcription Factor RelA

We have previously showed rTgPI-1 tolerogenic adjuvant properties in asthma treatment, turning it a promising candidate for allergen-specific immunotherapy. This therapy is an alternative treatment to control asthma that still presents several concerns related to its formulation. rTgPI-1 contains independent inhibitory domains able to inhibit trypsin and neutrophil elastase, both involved in asthma pathology.. In view of the need to design rational therapies, herein we investigated the contribution of the different inhibitory domains in rTgPI-1 therapeutic effectiveness.. BALB/c mice were rendered allergic by intraperitoneal OVA-alum sensitization and airway challenged. Once the asthmatic phenotype was achieved, mice were intranasally treated with OVA combined with the full-length recombinant protein rTgPI-1 or its truncated versions, Nt (containing trypsin-inhibitory domains) or Ct (containing neutrophil elastase-inhibitory domains). Afterward, mice were aerosol re-challenged.. Asthmatic mice treated with the neutrophil elastase- or the trypsin-inhibitory domains separately failed to improve allergic lung inflammation. Only when all inhibitory domains were simultaneously administered, an improvement was achieved. Still, a better outcome was obtained when mice were treated with the full-length rTgPI-1.. Adjuvant ability depends on the presence of all its inhibitory domains in a single entity, so it should be included in potential asthma treatment formulations as a full-length protein.

Topics: Adjuvants, Immunologic; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Leukocyte Elastase; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Serine Proteinase Inhibitors; Toxoplasma; Trypsin; Vaccination

Ganoderma, a fungal genus, is a traditional medicine with immuno-modulating effects. Asthma is an inflammatory disease of airways, and the main trigger of asthma is allergic inflammation. In this study, the effects of Ganoderma (an anti-inflammatory agent) given via oral administration (G/O) or intraperitoneal injection (G/IP) on asthma was evaluated. Forty BALB/c mice were divided into four groups, including the control, OVA-challenge, OVA-challenge + G/O, and OVA-challenge + G/IP. To determine AHR, the MCh challenge test was done. The levels of IL-1β, -4, -5, -6, -8, -10, -12, -13, -17, -25, -33, -38, Cys-LT, LTB4, and hydroxyproline were measured. Finally, lung histopathology was evaluated to determine eosinophilic inflammation, goblet cell hyperplasia, and mucus hyper-secretion. Treatment with G/O and G/IP could significantly reduce the levels of IL-1β, -5, -6, -8, -17, -25, -33, and -38; the levels of IL-4 and IL-13 had no significant changes, but the levels of IL-10 and IL-12 were enhanced. The mice treated with G/O and G/IP showed decreased levels of Cys-LT, LTB4, peribronchial and perivascular inflammation, but no significant changes were observed in AHR, hydroxyproline level, goblet cell hyperplasia, and mucus hyper-secretion. Ganoderma can be applied as an immunomodulatory and anti-inflammatory agent for managing asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Ganoderma; Hydroxyproline; Hyperplasia; Inflammation; Leukotriene B4; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Asthma is one of the most common chronic inflammatory diseases of the airways. Essential oil from Abies holophylla leaf (EOA) has been reported to have anti-inflammatory property. This study aimed to predict the inhibitory effect of EOA against asthma by network analysis and to confirm the underlying mechanism of EOA on airway inflammation.. The effects and underlying mechanisms of EOA on asthma were investigated by in silico network pharmacology and an experimental in vivo study.. To define the effectiveness of EOA on asthma, the network pharmacology was constructed using major components of EOA. EOA (0.0003 and, 0.03 v/v%) was aerosolized by nebulizer 3 times a week for 5 min for 7 weeks. After 3 weeks of treating the mice with EOA, asthma was induced by sensitizing them with ovalbumin (OVA) and PM10. The effects of EOA on the IL-17 related signaling pathway was confirmed using an asthmatic model.. The network analysis showed that EOA is highly associated with the IL-17-related signaling pathway. EOA inhibited respiratory epithelium hyperplasia, collagen deposition and goblet cell activation in the lung and trachea tissues. In addition, EOA reduced the number of eosinophils, lymphocytes and macrophages in BALF. Furthermore, in the asthmatic model of mice, we showed that EOA inhibited IL-17-related cytokines, increased Treg-related cytokines and decreased the TRAF6 and MAPK and, suppressed the nuclear transcriptional activities of NF-kB.. The network pharmacology and in vivo study indicated that EOA may have an inhibitory effect on airway inflammation in asthma exposure through the IL-17-related signaling pathway.

Topics: Abies; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Network Pharmacology; Oils, Volatile; Ovalbumin; Plant Leaves

Allergic asthma is one of the leading respiratory diseases with complex pathology. Attributes of vitexin, a trihydroxyflavone, has been studied to alleviate Th2 cytokines response in allergic asthma. However, its efficacy and underlying mechanism in mitigating allergic asthma particularly mediated by oxi-inflammatory stress, autophagy and apoptosis, yet to be delineated.. Present study aimed to decipher efficacy and governing molecular mechanism of vitexin in mitigating allergic asthma particularly mediated by vicious loop of oxi-inflammatory stress, autophagy and apoptosis.. To ascertain this, OVA-LPS induced mice model was used and protective attributes of vitexin for different mediators, pathological facets and sensing pathways of allergic asthma were evaluated.. Vitexin treatment remarkably inhibited OVA-LPS induced inflammatory cell infiltration, mast cell activation, alveolar collapse, congestion, fibrosis in lung architecture. These results were accompanied by suppression of immune cells hyperactivation, mucus secretion, goblet cell proliferation, persistent inflammation which were affirmed by alleviation in levels of IgE, Th1/Th2/Th17, IL-4/IFN-γ, chemokines, endopeptidases (MMP-1, MMP-13), oxidative effectors with concomitant increase in IL-15, IL-10, MMP-9 and MMP-3. Additionally, noticeable decline in p-connexin 43, p-c-Fos, TGF-β, Smad2/3/4, Caspase9/3, LC3A/B expression and upregulation in beclin-1, p62 co-localization and Bcl2/Bax indicate reversal of lung vascular permeability, mast cell degranulation, fibrosis, apoptosis, autophagosome impairment. Subsequent allergic inflammatory cascades analysis revealed p-NF-κB, p-PI3K, p-Akt, p-p38, p-Stat3, GATA3 upregulation and p-PTEN downregulation in sensitized mice, which were decisively counteracted by vitexin. In silico studies signified target specificity of vitexin with these proteins. Suppression in myeloid cells activation and enhancements of Tregs demonstrated immunomodulatory potential of vitexin in allergic airways.. Collectively, to our knowledge, this is the first report that confers vitexin meditated multi-faceted protective attribute in mitigation of allergic asthma that could be linked to its suppressive effects on vicious cycle of pathological process particularly regulated via oxi-inflammation, autophagy and apoptosis. Thus, signify vitexin as safe therapeutic strategy.

Topics: Animals; Apigenin; Asthma; Autophagy; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Homeostasis; Lung; Mice; Mice, Inbred BALB C; Myeloid Cells; Ovalbumin; Oxidation-Reduction

Sigma non-opioid intracellular receptor 1 (Sigma-1R) has been proven to play a major role in inflammation and structural remodeling. However, its role in airway inflammation and airway remodeling remains unclear. The purpose of this study aimed to explore the role and mechanism of Sigma-1R in airway remodeling and epithelial-mesenchymal transition (EMT) process in vivo and in vitro. We observed the decrease of Sigma-1R in lung tissue of asthma model. In the mouse model of allergic airway inflammation (AAI), Sigma-1R agonist RPE-084 significantly relieved airway inflammation and airway remodeling, while Sigma-1R antagonist BD1047 (B8562) had opposite effects. Further research showed that RPE-084 treatment increased the expression of pAMPK and inhibited the expression of CXCR4. Furthermore, RPE-084 treatment suppressed the levels of IL-4, IL-5, and IL-13 in BALF. We found that RPE-084 or Sigma-1R overexpression vector treatment regulated cell cycle and inhibited cell proliferation, migration, and EMT process in TGF-β1-induced 16HBE cells. Finally, we confirmed that AMP-activated protein kinase (AMPK) inhibitor compound C or CXCR4 agonist ATI-2341 both reversed the effects of Sigma-1R on TGF-β1-induced 16 HBE cells. In a word, our research shows that Sigma-1R is helpful to improve airway remodeling of asthma, and emphasizes a new candidate molecular for asthma treatment.

Topics: Airway Remodeling; AMP-Activated Protein Kinases; Animals; Asthma; Disease Models, Animal; Inflammation; Mice; Ovalbumin; Receptors, CXCR4; Receptors, sigma; Sigma-1 Receptor; Signal Transduction; Transforming Growth Factor beta1

Bacillus subtilis is an intestinal probiotic for immune homeostasis and its exopolysaccharide (EPS) is known to possess anti-inflammatory and antioxidant properties. The underlying mechanisms are not yet fully understood. In the present study, we investigated the effects of the EPS (50, 100, 200 mg/kg) on airway inflammation in asthmatic mice. Our results showed that EPS treatment of asthmatic mice significantly alleviated pathological damage in the lungs, remarkably decreased the counts of total inflammatory cells including lymphocytes, and eosinophils in the bronchoalveolar lavage fluid (BALF) and reduced indexes of oxidative damage. Moreover, the expression of type II T-helper cell (Th2) cytokines (interleukin- (IL)4 and -5) subsequent to EPS treatment was found to be dramatically down-regulated in a concentration-dependent manner. Additionally, the EPS treatment reduced JAK1, STAT6 and nuclear factor-κB (NF-κB) expression in the lungs of asthmatic mice. Taken together, these results suggest that the EPS from B. subtilis alleviates asthmatic airway inflammation, which involves the reduction in reactive oxygen species (ROS) and the down-regulation of the STAT6 and NF-κB inflammatory pathways, which can further reduce Th2 cytokine expression and eosinophilic inflammation. Thus, our findings provide a potential mechanism through which the EPS mitigates asthma, suggesting that the EPS could be a potential source of an anti-asthmatic drug.

Topics: Animals; Asthma; Bacillus subtilis; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Polysaccharides, Bacterial; STAT6 Transcription Factor

This study sought to examine whether ligustrazine was capable of inhibiting phosphodiesterase (PDE) activity and improving lung function in a rat model of asthma.. Rats were initially sensitized using ovalbumin (OVA) and then were challenged daily with aerosolized OVA beginning 14 days later (30 min/day) to generate a rat model of asthma. Changes in airway function following methacholine (MCh) injection were evaluated by monitoring lung resistance (. Ligustrazine suppresses airway inflammation and bronchial hyperresponsivity in this rat model system, and these changes are associated with decreased PDE expression at the protein and mRNA levels.

Topics: Airway Resistance; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Computational Biology; Disease Models, Animal; Immunoglobulin E; Lung; Male; Ovalbumin; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Pyrazines; Rats; Rats, Sprague-Dawley; Respiratory Hypersensitivity; RNA, Messenger

Tris (2-butoxyethyl) phosphate (TBEP) is an organophosphate flame retardant and used as a plasticizer in various household products such as plastics, floor polish, varnish, textiles, furniture, and electronic equipment. However, little is known about the effects of TBEP on the brain and behavior. We aimed to examine the effects of dietary exposure of TBEP on memory functions, their-related genes, and inflammatory molecular markers in the brain of allergic asthmatic mouse models. C3H/HeJSlc male mice were given diet containing TBEP (0.02 (TBEP-L), 0.2 (TBEP-M), or 2 (TBEP-H) μg/kg/day) and ovalbumin (OVA) intratracheally every other week from 5 to 11 weeks old. A novel object recognition test was conducted in each mouse at 11 weeks old. The hippocampi were collected to detect neurological, glia, and immunological molecular markers using the real-time RT-PCR method and immunohistochemical analyses. Mast cells and microglia were examined by toluidine blue staining and ionized calcium-binding adapter molecule

Topics: Animals; Asthma; Calcium-Binding Proteins; Dietary Exposure; Disease Models, Animal; Flame Retardants; Gene Expression Regulation; Male; Mast Cells; Memory; Mice; Mice, Inbred C3H; Microfilament Proteins; Microglia; Nerve Tissue Proteins; Organophosphorus Compounds; Ovalbumin; Oxidative Stress; Receptors, N-Methyl-D-Aspartate

The susceptibility to respiratory syncytial virus (RSV) infection in early life has been associated with a deficient T-helper cell type 1 (Th1) response. Conversely, healthy adults generally do not exhibit severe illness from RSV infection. In the current study, we investigated whether Th1 cytokine IFN-γ is essential for protection against RSV and RSV-associated comorbidities in adult mice. We found that, distinct from influenza virus, prior RSV infection does not induce significant IFN-γ production and susceptibility to secondary

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Coinfection; Comorbidity; Female; Inflammation; Interferon-gamma; Lung; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Respiratory Syncytial Virus Infections; Th1 Cells; Th2 Cells

Asthma, caused by chronic inflammation, is a common disease. Anthocyanins are involved in asthma treatment. This study explored the mechanism of anthocyanins on airway inflammation in asthmatic mice by regulating nuclear factor-κB (NF-κB) via the miR-138-5p/sirtuin-1 (SIRT1) axis.. The asthmatic mouse model was established by ovalbumin (OVA) induction and treated with anthocyanins or simultaneously injected with the lentivirus miR-138-5p mimic, followed by the measurement of lung inflammatory injury and IL-4, IL-5, IL-13, and IFN-γ levels in bronchoalveolar lavage fluid. Human bronchial epithelial (HBE) cells 16HBE14o-160 were induced by OVA to establish an asthmatic cell model, treated with anthocyanins and manipulated with miR-138-5p mimic and pcDNA3.1-SIRT1. The releases of inflammatory cytokines, the nuclear translocation of p-p65/p65 in the NF-κB pathway, and the levels of miR-138-5p and SIRT1 mRNA were detected.. In vivo experiments showed that anthocyanins could reduce the OVA-induced airway hyperresponsiveness and airway inflammation, improve the inflammatory infiltration and mucus in lung tissues, and diminish the miR-138-5p level in asthmatic mice. Infection with the miR-138-5p mimic averted the remission effect of anthocyanins in asthmatic mice. In vitro experiments showed that in HBE cells exposed to OVA, anthocyanins reduced the miR-138-5p level, increased the SIRT1 level, inhibited the release of inflammatory factors, and reduced the nuclear translocation of NF-κB p65. miR-138-5p targeted SIRT1. miR-138-5p overexpression partially reversed the therapeutic effect of anthocyanins, while SIRT1 overexpression antagonized the effect of miR-138-5p overexpression.. Anthocyanins inhibited the NF-κB pathway by regulating the miR-138-5p/SIRT1 axis, thus inhibiting airway inflammation in asthmatic mice.

Topics: Animals; Anthocyanins; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Mice; Mice, Inbred BALB C; MicroRNAs; NF-kappa B; Ovalbumin; Sirtuin 1

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hot Temperature; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; Th2 Cells

Cardiovascular disease remains the leading cause of death globally, and heart failure (HF) represents its terminal stage. Asthma, one of the most common chronic diseases, has been reported to be associated with an increased risk of cardiovascular disease. However, the link between asthma and HF has rarely been studied, and the possible mechanisms by which asthma affects HF are unclear. This study aimed to explore the influence of asthma on HF and the possible mechanisms. We analyzed data from the National Health and Nutrition Examination Survey and found a higher prevalence of HF among asthmatic individuals, and identified an independent association between HF and asthma. Subsequently, we produced mice with concurrent ovalbumin (OVA) sensitization-induced allergic asthma and angiotensin Ⅱ infusion-induced cardiac remodeling to explore the effect of asthma on cardiac remodeling in vivo. The results showed that OVA-induced asthma impaired heart function and aggravated cardiac remodeling in mice. We also found that OVA sensitization increased the expression levels of immunoglobulin E (IgE) in serum and IgE receptor (FcεR1) in the heart, and enhanced the activation of downstream signaling molecules of IgE-FcεR1 in the heart. Importantly, blockage of IgE-FcεR1 using FcεR1-deficient mice or an anti-IgE antibody prevented asthma-induced decline of cardiac function, and alleviated cardiac remodeling. These findings demonstrate the adverse effects of allergic asthma on the heart, and suggest the potential application of anti-IgE therapy in the treatment of asthma complicated with heart conditions.

Topics: Angiotensin II; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cardiovascular Diseases; Disease Models, Animal; Heart Failure; Immunoglobulin E; Mice; Mice, Inbred BALB C; Nutrition Surveys; Ovalbumin; Ventricular Remodeling

Aerva lanata Linn. (A. lanata) is traditionally used for cough, sore throat and asthma.. The aim of the present study was to investigate the immunomodulatory and anti-inflammatory potentials of A. lanata in allergic asthmatic mice.. BALB/c mice were administered with three different (methanol, n-hexane and ethyl acetate) extracts of A. lanata two weeks after immunization with ovalbumin and continued for 7 days. Inflammatory cells count was estimated in blood and broncho-alveolar lavage fluid (BALF). RT-PCR was used to find out mRNA expression levels of inflammatory mediators. GC-MS analysis was also carried out.. Among three extracts of A. lanata, ethyl acetate extract ameliorated (p < 0.001) count of inflammatory cells both blood and BALF remarkably. This study indicated that ethyl acetate extract of A. lanata lowered (p < 0.001) the level of inflammatory modulator TNF-α and IgE antibodies. A. lanata reduced (p < 0.001) interleukin 4, 5, 13 and enhanced (p < 0.001) expression levels of AQP1 and AQP5 in asthmatic mice. GC-MS analysis of ethyl acetate fraction indicated the presence of various anti-oxidant phyto-constituents. The groups treated with A. lanata improved inflammatory, goblet cells hyperplasia scoring and alveolar thickening.. The anti-asthmatic effect of A. lanata might be contributed by the suppression of edema, pro-inflammatory cytokines and IgE antibodies, and elevation of aquaporin expression levels, suggesting future study and clinical trials to propose it as a candidate to treat allergic asthma. The anti-oxidant phytochemicals present in A. lanata might be responsible for such potential.

Topics: Amaranthaceae; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antioxidants; Asthma; Cytokines; Immunomodulating Agents; Inflammation Mediators; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Solvents

Attributes of agnuside, a nontoxic, iridoid glycoside have been advocated for inflammatory disorders. However, information on its efficacy in alleviating allergic asthma largely remain ambiguous and yet to be deciphered. Present study aimed to assess efficacy of agnuside in targeting vicious circle of oxi-inflammation, autophagy and fibrosis, together with investigating its underlying molecular mechanism during OVA-LPS induced allergic asthma. Results revealed that agnuside showed prophylactic effect in assuaging asthmatic lung architecture impairment (p ≤ 0.01) as indicated by suppression of inflammatory cell infiltration, congestion, fibrosis, airway remodeling and alveolar collapse in OVA-LPS sensitized group. Decreased expression level (p ≤ 0.05) of allergic inflammatory mediators such as IgE, Th1/Th2, IL-4/IFN-γ, IL-4/IL-10, chemokines, endopeptidases and TGF-β, Smad2/4, Caspase9/3, connexin 43/50 observed in agnuside treatments. Analysis of redox molecular signaling cascade and autophagic proteins revealed concurrent upregulation in p-NF-κB, p-PI3K, p-Akt, p-p38, p-Stat3 activation, GATA3, LC3B expression and reduction in Bcl2/Bax, Beclin1 and p62 expression in sensitized mice (p ≤ 0.05) which were intensely counteracted by administration of agnuside. Suppression in myeloid cells activation and augmentation (p ≤ 0.001) of Tregs established modulatory attribute of agnuside for innate and adaptive immune response during allergic asthma. Collectively, these outcomes confer prophylactic attribute of agnuside and signify it as promising strategy to thwart allergic asthma.

Topics: Animals; Apoptosis; Asthma; Autophagy; Bronchoalveolar Lavage Fluid; Cell Lineage; Cytokines; Disease Models, Animal; Fibrosis; Glucosides; Homeostasis; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Ferroptosis was reported to be involved in the occurrence and development of asthma. However, the potential mechanism underlying the role of ferroptosis in asthma remains unclear. In this study, we established the mouse asthma model following the ovalbumin (OVA) method in C57BL/6 mice and the cell model with IL-13 induction in bronchial epithelial cells (BEAS-2B cells). Treatment of ferrostatin-1 (Ferr-1) and 3-methyladenine (3-MA) decreased iron deposition in IL-13-induced BEAS-2B cells and lung tissues of asthma mice, opposite to that in bronchoalveolar lavage fluid (BALF). Meanwhile, excessive lipid peroxidation asthma model

Topics: Adenine; Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cell Line; Cyclohexylamines; Cytokines; Disease Models, Animal; Epithelial Cells; Ferroptosis; Humans; Injections, Intraperitoneal; Interleukin-13; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Oxidative Stress; Phenylenediamines; Signal Transduction

To investigate the effect of eupatilin in asthma treatment, we evaluated its therapeutic effect and related signal transduction in OVA-induced asthmatic mice and LPS-stimulated RAW264.7 cells. The BALF was tested for changes in lung inflammatory cells. Th2 cytokines in the BALF and OVA-IgE in the serum were measured by ELISA. H&E and PAS staining were used to evaluate histopathological changes in mouse lungs. The key proteins NF-κB, MAPK, and Nrf2 in lung tissues were quantitatively analyzed by Western blotting. Finally, we evaluated the effect of eupatilin on cytokines and related protein expression in LPS-stimulated RAW 264.7 cells in vitro. In OVA-induced asthmatic mice, eupatilin reduced the numbers of inflammatory cells, especially neutrophils and eosinophils. Eupatilin also decreased the levels of IL-5, IL-13 in the BALF and OVA-IgE in the serum. Furthermore, eupatilin inhibited the activation of NF-κB and MAPK pathways and increased the expression of Nrf2 in OVA-induced asthmatic mice. In vitro, eupatilin significantly reduced LPS-stimulated NO, IL-6, and ROS production. Additionally, the NF-κB, MAPK, and Nrf2 protein expression in LPS-stimulated RAW264.7 cells was consistent with that in OVA-induced asthmatic lung tissues. In summary, eupatilin attenuated OVA-induced asthma by regulating NF-κB, MAPK, and Nrf2 signaling pathways. These results suggest the utility of eupatilin as an anti-inflammatory drug for asthma treatment.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Female; Flavonoids; Gene Expression Regulation; Lipopolysaccharides; MAP Kinase Signaling System; Mice; Molecular Structure; Neutrophils; NF-E2-Related Factor 2; NF-kappa B; Ovalbumin; RAW 264.7 Cells; Reactive Oxygen Species

Understanding the interaction between nanoparticles and immune cells is essential for the evaluation of nanotoxicity and development of nanomedicines. However, to date, there is little data on the membrane microstructure and biochemical changes in nanoparticle-loaded immune cells. In this study, we observed the microstructure of nanoparticle-loaded macrophages and changes in lipid droplets using holotomography analysis. Quantitatively analyzing the refractive index distribution of nanoparticle-loaded macrophages, we identified the interactions between nanoparticles and macrophages. The results showed that, when nanoparticles were phagocytized by macrophages, the number of lipid droplets and cell volume increased. The volume and mass of the lipid droplets slightly increased, owing to the absorption of nanoparticles. Meanwhile, the number of lipid droplets increased more conspicuously than the other factors. Furthermore, alveolar macrophages are involved in the development and progression of asthma. Studies have shown that macrophages play an essential role in the maintenance of asthma-related inflammation and tissue damage, suggesting that macrophage cells may be applied to asthma target delivery strategies. Therefore, we investigated the target delivery efficiency of gold nanoparticle-loaded macrophages at the biodistribution level, using an ovalbumin-induced asthma mouse model. Normal and severe asthma models were selected to determine the difference in the level of inflammation in the lung. Consequently, macrophages had increased mobility in models of severe asthma, compared to those of normal asthma disease. In this regard, the detection of observable differences in nanoparticle-loaded macrophages may be of primary interest, as an essential endpoint analysis for investigating nanomedical applications and immunotheragnostic strategies.

Topics: Animals; Asthma; Disease Models, Animal; Drug Delivery Systems; Feasibility Studies; Female; Gold; Lipopolysaccharides; Lung; Macrophages; Metal Nanoparticles; Mice; Nitric Oxide; Nitric Oxide Synthase Type II; Ovalbumin; RAW 264.7 Cells; Tissue Distribution; Tomography

Grape seed proanthocyanidin extract (GSPE) is effective in treating severe asthma (SA).. To examine the relationship between Nrf2-miR-29b axis and SA, and to detect whether preventive use of GSPE relieves SA via it.. SA group demonstrated significantly lower concentrations of Nrf2 protein, Nrf2 mRNA, and miR-29b than nSA group and control group. Conversely, higher levels of platelet derived growth factor C (PDGFC), phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1), and collagen type III alpha 1 (COL3A1) were measured in SA than in the other two groups. PDGFC, PIK3R1, and COL3A1 were the target genes of miR-29b. GSPE + DXM significantly elevated the expression of Nrf2 (+188%), Nrf2 mRNA (+506%), and miR-29b (+201%), and significantly reduced the expression of PDGFC (-72%), PIK3R1 (-40%), and COL3A1 (-65%) compared with OVA + LPS.. Nrf2-miR-29b axis is involved in the pathogenesis of SA. GSPE, as an adjuvant drug, maybe a potential therapeutic agent for SA.

Topics: Adult; Animals; Anti-Asthmatic Agents; Asthma; Case-Control Studies; Dexamethasone; Disease Models, Animal; Drug Therapy, Combination; Female; Gene Expression Regulation; Grape Seed Extract; Humans; Leukocytes, Mononuclear; Lipopolysaccharides; Male; Mice; Mice, Inbred BALB C; MicroRNAs; Middle Aged; NF-E2-Related Factor 2; Ovalbumin; Proanthocyanidins; Severity of Illness Index

Different endotypes of asthma were described in human. Atopic asthma is a T-helper 2 (Th2)-mediated disease consisting mainly of an eosinophilic inflammation in the airways. Other endotypes show neutrophilic inflammation of the airways that is probably based on a Th17 response. There are several mouse models described in the literature to study the Th2 polarized eosinophilic disease, however, only a few models are available which characterize the neutrophilic endotype. The aim of this study was to compare both endotypes in relation to the severity of the allergen-induced inflammation. Groups of either Balb/c or DO11.10 mice were sensitized with ovalbumin (OVA) adsorbed to aluminum hydroxide. Mice were subsequently challenged with OVA for different periods of time. They were evaluated for airway hyperreactivity (AHR), cytokine production, airway inflammation, and remodeling of the airways. As expected, Balb/c mice developed a Th2 response with AHR, eosinophilic airway inflammation, and allergen-specific IgE and IgG1. By contrast DO11.10 mice showed a mixed Th1/Th17 response with strong neutrophilic airway inflammation, IgG2a, but only limited induction of AHR. While Balb/c mice showed remodeling of the airways with subepithelial fibrosis and goblet cell metaplasia, airway remodeling in DO11.10 mice was marginal. Both airway inflammation and remodeling resolved after prolonged periods of challenge in both models. In conclusion, strong allergen-induced airway remodeling in mice seems to be triggered by the specific conditions arising from infiltration with eosinophilic granulocytes in the lung. A Th1/Th17 response leading to neutrophilic inflammation does not seem to be sufficient to induce pronounced airway remodeling.

Topics: Airway Remodeling; Allergens; Animals; Asthma; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Asthma is the leading inflammatory disease of the airways with inadequate therapeutic options. 'Malla Sindoor' (MS) is a metal-based ethnomedicinal formulation that has been prescribed in the ancient traditional medicinal system for treating chronic inflammations.. Here, we validated the anti-inflammatory and anti-asthmatic properties of traditional metallic medicine MS in asthmatic mice model and in LPS stimulated human monocytic THP-1 cells, by examining the relevant cellular, biochemical and molecular intermediates.. Scanning Electron Microscope (SEM), Electron Dispersive X-ray (EDX), and X-Ray Diffraction (XRD) were performed to characterize MS particles. Allergic asthma was induced in Balb/c mice through intraperitoneal ovalbumin (OVA) injection. Experimental groups include, normal control, disease control, Dexamethasone (2 mg/kg) and three MS treated groups: 4.3 mg/kg, 13 mg/kg, and 39 mg/kg. Quantitative PCR, inflammatory cytokines and anti-oxidant enzymes, and histological analysis were performed, in the treated mice and LPS stimulated human monocytic THP-1 cells for determining the MS efficacy.. SEM image analysis showed the MS to be heterogenous in shape with a particle size distribution between 100 nm-1 μm. Elemental composition showed the presence of mercury (Hg), arsenic (As), and sulphur (S) along with other elements in the forms of mercury sulfide, arsenic trioxide, and their alloy crystals. OVA-challenge of the Balb/c mice resulted in the development of overt pathological features for allergic asthma including smooth muscle thickening and collagen deposition. Mice receiving MS-exhibited alleviation of allergic asthma features. BAL fluid analysis showed a decrease in the total cell count and decreases in neutrophils, monocytes, lymphocytes, and eosinophils. Further, the stimulated levels of interleukin (IL)-1β, -6, and TNF-α cytokines and antioxidant levels were also reduced upon MS-treatment. At the molecular level, MS-treatment reduced stimulated mRNA expression levels for IL-4, -5, -10, -13, -33, and IFN-γ cytokines. Histological analysis following MS-treatment of OVA-stimulated mice lungs showed a reduction in mucus accumulation in airways, decreases in peribronchial collagen deposition, bronchial smooth muscle thickening, and attenuation of inflammatory cell infiltration. In addition, under in-vitro conditions, MS-treatment attenuated the LPS induced secretion of IL-1β, -6, and TNF-α from THP-1 cells.. Collectively, the results suggest that MS acts as an effective anti-asthmatic and anti-inflammatory agent, by regulating various cellular, biochemical and molecular intermediates.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antioxidants; Asthma; Cytokines; Disease Models, Animal; Lipopolysaccharides; Medicine, Traditional; Mercury; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Pneumonia; Tumor Necrosis Factor-alpha

Mounting evidence suggests that nematode infection can protect against disorders of immune dysregulation. Administration of live parasites or their excretory/secretory (ES) products has shown therapeutic effects across a wide range of animal models for immune disorders, including asthma. Human clinical trials of live parasite ingestion for the treatment of immune disorders have produced promising results, yet concerns persist regarding the ingestion of pathogenic organisms and the immunogenicity of protein components. Despite extensive efforts to define the active components of ES products, no small molecules with immune regulatory activity have been identified from nematodes. Here we show that an evolutionarily conserved family of nematode pheromones called ascarosides strongly modulates the pulmonary immune response and reduces asthma severity in mice. Screening the inhibitory effects of ascarosides produced by animal-parasitic nematodes on the development of asthma in an ovalbumin (OVA) murine model, we found that administration of nanogram quantities of ascr#7 prevented the development of lung eosinophilia, goblet cell metaplasia, and airway hyperreactivity. Ascr#7 suppressed the production of IL-33 from lung epithelial cells and reduced the number of memory-type pathogenic Th2 cells and ILC2s in the lung, both key drivers of the pathology of asthma. Our findings suggest that the mammalian immune system recognizes ascarosides as an evolutionarily conserved molecular signature of parasitic nematodes. The identification of a nematode-produced small molecule underlying the well-documented immunomodulatory effects of ES products may enable the development of treatment strategies for allergic diseases.

Topics: Animals; Asthma; Disease Models, Animal; Host-Pathogen Interactions; Hypersensitivity; Inflammation; Mice; Mice, Inbred BALB C; Nematoda; Ovalbumin; Small Molecule Libraries; Trachea

Asthma is a chronic inflammation of pulmonary airways associated with bronchial hyper-responsiveness. The study was aimed to validate the folkloric use of Polystichum braunii (PB) against ovalbumin (OVA)-induced asthmatic and chemical characterization OF both extracts. Allergic asthma was developed by intraperitoneal sensitization with an OVA on days 1 and 14 followed by intranasal challenge. Mice were treated with PB methanolic (PBME) and aqueous extract (PBAE) orally at 600, 300, and 150 mg/kg and using dexamethasone (2 mg/kg) as standard from day 15 to 26. High performance liquid chromatography-diode array detector analysis revealed the presence of various bioactive compounds such as catechin, vanillic acid, and quercetin. The PBME and PBAE profoundly (p < 0.0001-0.05) declined immunoglobulin E level, lungs wet/dry weight ratio, and total and differential leukocyte count in blood and bronchial alveolar lavage fluid of treated mice in contrast to disease control. Histopathological examination showed profoundly decreased inflammatory cell infiltration and goblet cell hyperplasia in treated groups. Both extracts caused significant (p < 0.0001-0.05) diminution of IL-4, IL-5, IL-13, IL-6, IL-1β, TNF-α, and NF-κB and upregulation of aquaporins (1 and 5), which have led to the amelioration of pulmonary inflammation and attenuation of lung edema in treated mice. Both extracts profoundly (p < 0.0001-0.05) restored the activities of SOD, CAT, GSH and reduced the level of MDA dose dependently. Both extracts possessed significant anti-asthmatic action mainly PBME 600 mg/kg might be due to phenols and flavonoids and could be used as a potential therapeutic option in the management of allergic asthma.

Topics: Animals; Anti-Asthmatic Agents; Aquaporins; Asthma; Biomarkers; Cytokines; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Plant Extracts; Polystichum; Pulmonary Edema

Control groups received treatment with normal saline or dexamethasone (2 mg/kg) on the same day. We assessed. MCHA significantly improved airway hyperresponsiveness near baseline levels. MCHA administration significantly improved airway and lung inflammation, demonstrated by decreased total and inflammatory cells in BAL, lower levels of IL-5 and IL-13 in lung homogenate, and fewer inflammatory cells in lung tissue. Additionally, MCHA significantly diminished goblet cells in lung tissue.. Administration of a hydroethanolic extract of

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Momordica charantia; Ovalbumin

Asthma is a chronic respiratory disease highly prevalent worldwide. Recent studies have suggested a role for microbiome-associated gut-lung axis in asthma development. In the current study, we investigated if Resveratrol (RES), a plant-based polyphenol, can attenuate ovalbumin (OVA)-induced murine allergic asthma, and if so, the role of microbiome in the gut-lung axis in this process. We found that RES attenuated allergic asthma with significant improvements in pulmonary functions in OVA-exposed mice when tested using plethysmography for frequency (F), mean volume (MV), specific airway resistance (sRaw), and delay time(dT). RES treatment also suppressed inflammatory cytokines in the lungs. RES modulated lung microbiota and caused an abundance of A

Topics: Animals; Asthma; Disease Models, Animal; Inflammation; Lung; Mice; Microbiota; Ovalbumin; Resveratrol

Virus-induced asthma exacerbation is a health burden worldwide and lacks effective treatment. To better understand the disease pathogenesis and find novel therapeutic targets, we established a mouse model of steroid (dexamethasone (DEX)) resistant asthma exacerbation using ovalbumin (OVA) and influenza virus (FLU) infection. Using liquid chromatography-tandem mass spectrometry (LC-MC/MS), we performed a shotgun proteomics assay coupled with label-free quantification to define all dysregulated proteins in the lung proteome of asthmatic mice. Compared to control, 71, 89, and 30 proteins were found significantly upregulated by at least two-fold (p-value ≤ 0.05) in OVA-, OVA/FLU-, and OVA/FLU/DEX-treated mice, respectively. We then applied a Z-score transformed hierarchical clustering analysis and Ingenuity Pathway Analysis (IPA) to highlight the key inflammation pathways underlying the disease. Within all these upregulated proteins, 64 proteins were uniquely highly expressed in OVA/FLU mice compared to OVA mice; and 11 proteins were DEX-refractory. IPA assay revealed two of the most enriched pathways associated with these over-expressed protein clusters were those associated with MHC class I (MHC-I) antigen-presentation and interferon (IFN) signaling. Within these pathways, signal-transducer-and-activator-of-transcription-1 (STAT1) protein was identified as the most significantly changed protein contributing to the pathogenesis of exacerbation and the underlying steroid resistance based on the label-free quantification; and this was further confirmed by both Parallel Reaction Monitoring (PRM) proteomics assay and western blots. Further, the pharmacological drug Fludarabine decreased STAT1 expression, restored the responsiveness of OVA/FLU mice to DEX and markedly suppressed disease severity. Taken together, this study describes the proteomic profile underpinning molecular mechanisms of FLU-induced asthma exacerbation and identifies STAT1 as a potential therapeutic target, more importantly, we provided a novel therapeutic strategy that may be clinically translated into practice.

Topics: Animals; Asthma; Cytokines; Mice; Mice, Inbred BALB C; Ovalbumin; Proteomics; Steroids; Vidarabine

Topics: Acupuncture Points; Animals; Asthma; Bronchoalveolar Lavage Fluid; Catgut; Cyclooxygenase 2; Cytokines; Disease Models, Animal; Female; Immunity, Innate; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Asthma is a common respiratory disease, and immune system dysregulation has direct relevance to asthma pathogenesis. Probiotics and prebiotics have immunomodulatory effects and can regulate immune responses and may attenuate allergic reactions. Therefore, in this study, we explored the role of probiotics and prebiotics in regulating acute airway inflammation and the TLR4/NF-kB pathway. Allergic asthma model of BALB/c mice was produced and treated with probiotics (LA-5, GG, and BB-12) and prebiotics (FOS and GOS). Then AHR, BALF cells count, EPO activity, IL-4, 5, 13, 17, 25, 33, as well as IFN-γ, total and OVA-specific IgE, IgG1, Cys-LT, LTB4, LTC4, and TSLP levels were measured. Also, the GTP/GOT assay was performed and gene expression of Akt, NLR3, NF-kB, PI3K, MyD88, TLR4, CCL11, CCL24, MUC5a, Eotaxin, IL-38, and IL-8 were determined. Finally, lung histopathological features were evaluated. Treatment with probiotics could control AHR, eosinophil infiltration to the BALF and reduce the levels of immunoglobulins, IL-17, GTP and also decrease mucus secretion, goblet cell hyperplasia, peribronchial and perivascular inflammation and also, EPO activity. It could reduce gene expression of TLR4 and CCL11. On the other hand, IL-38 gene expression was increased by both probiotic and prebiotic treatment. Treatment with probiotics and prebiotics could control levels of IL-4, 5, 13, 25, 33, leukotrienes, the gene expression of AKT, NLR3, NF-κB, MyD88, MUC5a. The prebiotic treatment could control peribronchial inflammation and PI3K gene expression. Both of the treatments had no significant effect on the GOT, TSLP and IL-8, eotaxin and CCL24 gene expression. Probiotics and prebiotics could induce tolerance in allegro-inflammatory reactions and alter immune responses in allergic conditions. Probiotics could also modulate cellular and humoral immune responses and prevent allergic disorders.

Topics: Animals; Asthma; Inflammation; Lung; Mice; NF-kappa B; Ovalbumin; Pneumonia; Prebiotics; Probiotics; Signal Transduction; Toll-Like Receptor 4

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humans; Lung; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Transcriptome

Asthma is a common chronic airway inflammatory disease, lacking effective therapeutic approaches. Magnesium isoglycyrrhizinate (MgIG) is an anti-inflammatory drug for treating chronic inflammation. However, it is still ambiguous whether MgIG can function in allergy induced asthma. In this study, we investigated the anti-inflammation effect of MgIG in mice with allergy induced asthma and explored the underlying mechanisms.. Mouse asthma model was established with ovalbumin (OVA) sensitization and challenge. Subsequently, mice sensitized with OVA were randomly assigned into fourgroups: asthma model group (MDL), dexamethasone group (DXM), MgIG group (MgIG), and normal mice were used as normal control (CON). The mice in MgIG, MDL were given 0.2 mg/mL MgIG solution by atomization inhalation for 30 min before 1% (w/v) OVA challenge. At the completion of model establishment and drug treatment, cells in bronchoalveolar lavage fluid were classified, inflammatory factors in serum were determined, histopathological analysis was performed by H&E staining, and expression of MUC5AC, NLRP3, and cleaved caspase-1 in the lung tissue was also determined by immunohistochemistry and western blotting, respectively.. In comparison to MDL group, MgIG treatment could significantly inhibit the recruitment of white blood cells, neutrophils, and eosinophils in BALF, reduced the production of IL-6, TNF-α, and IgE in serum, and reduced mucus secretion and the infiltration of inflammatory cells. Also, an increase of NLRP3 and Caspase-1 protein levels were suppressed by MgIG treatment.. Our study findings support that nebulizer inhalation of MgIG as an effective therapy in treating the allergy induced asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Caspases; Disease Models, Animal; Hypersensitivity; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Saponins; Triterpenes

Cigarette smoke (CS) is a common environmental irritant and a risk factor for asthma, as it induces as well as aggravates asthmatic attacks. The injured airway epithelial tight junctions (TJs) aggravate asthma. CS can aggravate asthma by activating the transient receptor potential ankyrin A1 (TRPA1) channel and enhancing TJs destruction. Houpo Mahuang decoction (HPMHD) is a classic traditional Chinese prescription for the treatment of asthma. However, its underlying action mechanism is unclear.. The present study aimed to evaluate the effect of HPMHD on the asthma phenotype and the regulation of TRPA1 and TJs in a CS-induced mouse model of aggravated asthma.. Under optimized chromatographic and mass spectrometry conditions, the ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) technique was used to detect and analyze the major chemical components of HPMHD. C57BL/6 female mice were randomly divided into seven groups, viz, normal saline (NS) group, ovalbumin (OVA) + CS group, dexamethasone group, HPMHD high-dose group and low-dose groups, n-butanol extract group, and ethyl acetate extract group, with 10 mice in each group. OVA sensitization and challenge, and CS exposure were used to establish the aggravated asthma model. As the main indices to evaluate the protective effect of HPMHD, the eosinophils count in peripheral blood, percentages of inflammatory cells classified and the levels of interleukin (IL)-4, IL-5, IL-13 in the bronchoalveolar lavage fluid (BALF), airway responsiveness enhanced pause (Penh), and changes in lung histopathology were determined and compared among the groups. The mRNA and protein expression of TRPA1 and TJs in lung tissue was also examined.. Using UPLC-QTOF-MS, the chemical components of HPMHD, including ephedrine, pseudoephedrine, laetrile, and amygdalin amide, were identified by 51 signal peaks. Compared with those in the NS group, the eosinophil number in the peripheral blood and the eosinophils and neutrophils percentages in BALF of the OVA + CS group were remarkably increased. Following the inhalation of 50 μl of acetylcholine chloride (ACH) at doses of 25 and 50 mg/mL, the Penh increased significantly (p < 0.01). Moreover, in the OVA + CS group, hematoxylin and eosin (H&E) staining of lung tissue showed a significant number of infiltrated inflammatory cells, increased mucus secretion in the lumen, damaged bronchial mucosa, increased thickness of tracheal wall, and increased score of lung damage (p < 0.01). The IL-4/5/13 levels were also remarkably increased (p < 0.01). The protein as well as gene expression of both ZO-1 and occludin decreased markedly in the lung tissue, while the expression of TRPA1 and claudin-2 was increased (p < 0.05, p < 0.01). Next, the OVA + CS group and the treatment groups were compared. The inflammatory cells, Penh value, and levels of IL-4/5/13 were significantly reduced, and less lung injury was observed in the treatment groups. The gene and protein levels of TRPA1 and TJs were corrected (p < 0.05, p < 0.01); the effects on the HPMHD high-dose and ethyl acetate extract groups were particularly remarkable.. HPMHD reduced airway hyperresponsiveness, inflammatory cell recruitment and Th2 cytokine secretion in CS-induced aggravated asthma mice, in a manner potentially dependent on regulation of the expression of TRPA1 and TJ proteins. Both the n-butanol and ethyl acetate extracts contained the active ingredients, especially the ethyl acetate extract.

Topics: 1-Butanol; Animals; Ankyrins; Asthma; Bronchoalveolar Lavage Fluid; Cigarette Smoking; Disease Models, Animal; Drugs, Chinese Herbal; Female; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Tight Junctions; Transient Receptor Potential Channels; TRPA1 Cation Channel

Progressive loss of effector functions, especially IFN-γ secreting capability, in effector memory CD8

Topics: Animals; Asthma; CD8-Positive T-Lymphocytes; Immune Tolerance; Immunologic Memory; Interferon-gamma; Mice; Ovalbumin

In this study, we sought to investigate the prospective role of circ 001372 in modifying inflammation in ovalbumin-induced asthma. In the vivo model of asthma, the serum of circ 001372 was reduced. Down-regulation of circ 001372 increased inflammation reaction (TNF-α, IL-1β, IL-6, and IL-18) and induced COX-2 and iNOS protein expression in vitro model through activation of NFAT5 and suppression of Sirt1. Up-regulation of circ 001372 decreased inflammation reaction (TNF-α, IL-1β, IL-6, and IL-18) in vitro model through inactivation of NFAT5 and induction of Sirt1 by miRNA-128-3p. The miRNA-128-3p lowered the effects of circ 001372 on inflammation in vitro model. The Sirt1 inhibitor reduced the effects of circ 001372 on inflammation in vitro model. Our results revealed the serum of circ 001372 against inflammation in ovalbumin-induced asthma through Sirt1/NFAT5 by miRNA-128-3p.

Topics: Asthma; Humans; Inflammation; Interleukin-18; Interleukin-6; MicroRNAs; Ovalbumin; RNA, Circular; Signal Transduction; Sirtuin 1; Transcription Factors; Tumor Necrosis Factor-alpha

Alkyl organophosphate flame retardants (OPFRs), tri-n-butyl phosphate (TnBP) and tris(2-butoxyethyl) phosphate (TBOEP), are ubiquitously detected in indoor and outdoor environments and their inhalation may result in lung damage. This study examined pulmonary toxicity after exposure to TnBP or TBOEP and investigated aggravation of inflammation and immunoreaction by TnBP in an ovalbumin (OVA)-induced mice model. Transcriptomics were used to further reveal the underlying mechanism. Exposure to TnBP or TBOEP resulted in pathological damage, including edema and thickened alveolar septum. In comparison with the control, enhanced levels of superoxide dismutase (SOD) (p < 0.01 in TnBP (High) group and p < 0.05 in TBOEP (High) group), glutathione peroxidase (GSH-px) (p < 0.05), malondialdehyde (MDA) (p < 0.01), and cytokines under a dose-dependent relationship were noted, and the expression of the Fkbp5/Nos3/MAPK/NF-кB signaling pathway (p < 0.01) was upregulated in the TnBP and TBOEP groups. Moreover, the combined exposure of TnBP and OVA exacerbated the allergic inflammatory response, including airway hyperresponsiveness, leukocytosis, cellular exudation and infiltration, secretion of inflammatory mediators, and higher expression of IgE (p < 0.01). Transcriptomics results demonstrated that the PI3K/Akt/NF-кB signal pathway was involved in TnBP-aggravated asthmatic mice. Exposure to TnBP or TBOEP resulted in oxidative damage and leukocyte-induced lung injury. TnBP can further facilitate OVA-induced asthma through an inflammatory response. This study is the first to reveal the pulmonary toxicity and potential mechanism induced by OPFRs through an in-vivo model.

Topics: Animals; Asthma; Flame Retardants; Mice; NF-kappa B; Organophosphates; Ovalbumin; Phosphatidylinositol 3-Kinases; Pneumonia; Signal Transduction

Topics: Animals; Asthma; Epithelial Cells; Epithelial-Mesenchymal Transition; Humans; Mice; Ovalbumin; Receptors, Tumor Necrosis Factor, Member 25; Tumor Necrosis Factor Ligand Superfamily Member 15

Bronchial asthma (BA) is a heterogeneous chronic inflammatory disease of the airways. The majority of patients with mild to moderate BA develop Th2-biased eosinophilic pulmonary inflammation and respond well to corticosteroid treatment. However up to 10% of BA patients develop severe pathology, which is associated with neutrophilic inflammation and resistant to conventional corticosteroid therapy. Contrary to eosinophil-predominant airway inflammation neutrophilic BA is developed through Th1- and Th17-immune responses. However, the etiology of corticoid insensitive neutrophilic BA is still remains unclear. Therefore, in the current study we developed a mouse model of BA with predominant neutrophilic rather than eosinophilic pulmonary inflammation. BALB/c mice were immunized with the mixture of the ovalbumin allergen and Freund's adjuvant, followed by aerosol challenge with the same allergen mixed with E. coli lipopolysaccharide. As a result, mice developed the main BA manifestations: production of allergen specific IgE, development of airway hyperreactivity, airway remodeling and pulmonary neutrophilic inflammation. Moreover, this pathology developed through Th1- and Th17-dependent mechanisms and mice with induced neutrophilic BA phenotype responded poorly to dexamethasone treatment, that coincide to clinical observations. The established mouse model could be useful both for studying the pathogenesis and for testing novel approaches to control neutrophilic BA.

Topics: Adrenal Cortex Hormones; Allergens; Animals; Asthma; Bronchial Hyperreactivity; Disease Models, Animal; Escherichia coli; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Pneumonia; Steroids

Asthma is a respiratory disease characterized by heterogeneous chronic airway inflammation. Activation of nucleotide-binding oligomerization domain-like receptor pyrin domain containing 3 (NLRP3) inflammasome is involved in the development of many pulmonary inflammatory diseases. The role and regulatory mechanism of carbenoxolone (CBX) in ovalbumin (OVA)-induced asthma models are not fully clear. Therefore, the study investigated whether CBX ameliorates airway inflammation and remodeling, as well as its mechanism in OVA induced-inflammation in mice. Wright-Giemsa staining was used to count inflammatory cells in bronchoalveolar lavage fluid (BALF). The level of inflammatory cells infiltration, mucus cell proliferation, and collagen deposition in lung tissue were separately assessed by hematoxylin and eosin, periodic acid-Schiff, and Masson trichrome staining, respectively. Airway resistance (AR) was measured by non-invasive airway system. Immunohistochemical assay was used to observe NLRP3 expression area. The expression of nuclear factor-kappaB (NF-κB), p-NF-κB, inhibitor of kappaB (IκB)-α, p-IκB-α, NLRP3, pro-caspase-1, caspase-1, and interleukin (IL)-1β in lung tissue were measured using quantitative real-time PCR or Western blotting. Our results showed that CBX can significantly attenuate the leukocyte count and the percentage of eosinophils and neutrophils in the BALF, peribronchial inflammation, airway mucus secretion, collagen deposition area, and AR in OVA-induced airway inflammation. In addition, the expression of p-NF-κB, p-IκB-α, NLRP3 and related factors were dramatically alleviated after CBX treatment. These data suggest that CBX has a significant protective effect on allergic airway inflammation by suppressing the activation of NLRP3 inflammasome through NF-κB pathway in asthmatic mice.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Carbenoxolone; Caspase 1; Disease Models, Animal; Inflammasomes; Inflammation; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; NF-KappaB Inhibitor alpha; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin

The purpose of this experiment is to find out the function of Vitamin D (VD) in airway inflammation in asthmatic guinea pigs by regulating mammalian target of rapamycin (mTOR)-mediated autophagy. A total of 40 male guinea pigs were randomly assigned into the Con group, the ovalbumin (OVA)-sensitized group, the VD group, the VD + dimethyl sulfoxide group, and the VD + rapamycin (mTOR inhibitor) group. Then, serum from all groups was harvested for the measurement of immunoglobulin E (IgE), interleukin (IL)-4, and IL-5 levels. Next, bronchoalveolar lavage fluid was collected for cell counting. Moreover, lung tissues were extracted to assess levels of p-mTOR and autophagy factors (LC3B, Beclin1, Atg5, and P62). Compared with the Con group, the OVA group showed elevated levels of IgE, IL-4, and IL-5, increased contents of eosinophils, neutrophil, and lymphocytes, and declined monocytes. And the VD group improved inflammatory reactions in the guinea pigs. Besides, the OVA group showed lower levels of p-mTOR and P62 and higher autophagy levels than the Con group, while the VD group had opposite results. Rapamycin annulled the suppressive role of VD to airway inflammation in asthmatic guinea pigs. VD might inhibit OVA-induced airway inflammation by inducing mTOR activation and downregulating autophagy in asthmatic guinea pigs.

Topics: Animals; Asthma; Autophagy; Disease Models, Animal; Female; Guinea Pigs; Immunoglobulin E; Inflammation; Interleukin-5; Lung; Male; Mammals; Ovalbumin; Sirolimus; TOR Serine-Threonine Kinases; Vitamin D

Asthma currently affects more than 339 million people worldwide. In the present preliminary study, we examined the efficacy of a new, inhalable soluble epoxide hydrolase inhibitor (sEHI), 1-trifluoromethoxyphenyl-3-(1-propionylpiperidin-4-yl) urea (TPPU), to attenuate airway inflammation, mucin secretion, and hyper-responsiveness (AHR) in an ovalbumin (OVA)-sensitized murine model. Male BALB/c mice were divided into phosphate-buffered saline (PBS), OVA, and OVA+TPPU (2- or 6-h) exposure groups. On days 0 and 14, the mice were administered PBS or sensitized to OVA in PBS. From days 26-38, seven challenge exposures were performed with 30 min inhalation of filtered air or OVA alone. In the OVA+TPPU groups, a 2- or 6-h TPPU inhalation preceded each 30-min OVA exposure. On day 39, pulmonary function tests (PFTs) were performed, and biological samples were collected. Lung tissues were used to semi-quantitatively evaluate the severity of inflammation and airway constriction and the volume of stored intracellular mucosubstances. Bronchoalveolar lavage (BAL) and blood samples were used to analyze regulatory lipid mediator profiles. Significantly (p < 0.05) attenuated alveolar, bronchiolar, and pleural inflammation; airway resistance and constriction; mucosubstance volume; and inflammatory lipid mediator levels were observed with OVA+TPPU relative to OVA alone. Cumulative findings indicated TPPU inhalation effectively inhibited inflammation, suppressed AHR, and prevented mucosubstance accumulation in the murine asthmatic model. Future studies should determine the pharmacokinetics (i.e., absorption, distribution, metabolism, and excretion) and pharmacodynamics (i.e., concentration/dose responses) of inhaled TPPU to explore its potential as an asthma-preventative or -rescue treatment.

Topics: Aerosols; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Epoxide Hydrolases; Humans; Inflammation; Lipids; Male; Mice; Mice, Inbred BALB C; Ovalbumin

Asthma is a chronic inflammatory disease of the respiratory system. Maresin-2 (MaR2) is biosynthesized from docosahexaenoic acid (DHA) by macrophages, display strong anti-inflammatory and pro-resolving activity. To investigate the therapeutic effect and mechanism of MaR2 on asthmatic mice induced by ovalbumin (OVA) in conjunction with the adjuvant aluminum hydroxide. Twenty four female BALB/c mice were randomly divided into control, OVA, OVA + MaR2, and OVA + dexamethasone (Dexa) groups. MaR2 or Dexa were given as a treatment for OVA-induced asthma. Serum, bronchoalveolar alveolar lavage fluid (BALF) and lung tissue were collected for further analysis. The Pathological changes of lung tissue, proportion of inflammatory cells in BALF, levels of inflammatory cytokines in BALF or serum, oxidative stress indices, and the protein concentration of ASC, MPO, Ly-6G, ICAM-1, NLRP3 and Caspase-1 in lung tissues were evaluated. Compared with the OVA group, both OVA + MaR2 and OVA + Dexa group had reduced inflammation and mucus secretion in lung tissue, number of inflammatory cells in BALF, levels of related inflammatory cytokines in serum or BALF, and expressions of ASC, MPO, Ly-6G, ICAM-1, NLRP3 and Caspase-1 proteins in lung tissue. In addition, the oxidative stress was alleviated as indicated by decreased MDA, and elevated SOD and GSH. MaR2 has an obvious protective effect on OVA-induced bronchial asthma in mice, in a similar manner as Dexa. The mechanism may be related to the inhibition of the Th2 type immune response, the NLRP3 inflammasome activation and oxidative stress.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Caspases; Cytokines; Disease Models, Animal; Docosahexaenoic Acids; Female; Immunity; Inflammasomes; Inflammation; Intercellular Adhesion Molecule-1; Lung; Mice; Mice, Inbred BALB C; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Oxidative Stress

Dioscin is a natural product that possesses protective effects on multiple chronic injuries, but its effects on asthma are not fully understood. Herein, we evaluated its effects on asthmatic mice established by ovalbumin (OVA) sensitization and challenges and further explored the mechanism. Inflammatory cells in bronchoalveolar lavage fluids (BALFs) were analyzed using Diff-Quik staining. OVA-specific immunoglobulin E (IgE)/IgG1 in serum and inflammatory cytokines (interleukin 4[IL-4], IL-5, IL-13, and tumor necrosis factor-α) in BALFs and lung tissues were measured using Enzyme-Linked Immunosorbent Assay Kits. Hematoxylin and eosin, periodic acid-Schiff, and immunohistochemistry staining showed histopathological changes in lung tissues. Epithelial-mesenchymal transition (EMT) in human bronchial epithelial (16HBE) cells was assessed by immunofluorescence staining. Hydroxyproline content was used to evaluate collagen deposition. Polymerase chain reaction and Western blot were performed to measure messenger RNA and protein expression. We found that dioscin treatment (particularly at the dose of 80 mg/kg) significantly inhibited pulmonary inflammation in asthmatic mice, as evidenced by the decreased serum OVA-specific IgE/IgG1 and the reduced inflammatory cells and cytokines in BALFs and lung tissues. Moreover, dioscin effectively ameliorated the goblet cell hyperplasia, mucus hypersecretion, collagen deposition, and smooth muscle hyperplasia in the airways of asthmatic mice. Mechanistically, dioscin restrained the activated TGF-β1/Smad2/3 and protein kinase B (AKT) signal pathways in lung tissues and potently reversed the TGF-β1-induced EMT and phosphorylation of Smad2/3 and AKT in 16HBE cells. Collectively, dioscin displayed protective effects on OVA-induced asthmatic mice via adjusting TGF-β1/Smad2/3 and AKT signal pathways, supporting the fact that dioscin could be a candidate for chronic asthma prevention in the future.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Collagen; Diosgenin; Disease Models, Animal; Humans; Hyperplasia; Immunoglobulin E; Immunoglobulin G; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Proto-Oncogene Proteins c-akt; Smad2 Protein; Smad3 Protein; Transforming Growth Factor beta1

Ferroptosis is closely associated with respiratory diseases; however, the relationship between ferroptosis and neutrophilic asthma remains unknown. This study investigated whether Liproxstatin-1 (Lip-1) affects the progression of neutrophilic asthma by inhibiting ferroptosis and inflammatory response, while dissecting the underlying molecular mechanisms.. The bronchial epithelial cells (16HBE and BEAS-2B) were administered with lipopolysaccharide (LPS) and interleukin-13 (IL-13) to generate a cell injury model. This cell model was employed to examine the effect of Lip-1 on airway epithelial-associated inflammation and ferroptosis as well as the underlying molecular mechanism. Meanwhile, we evaluated the effects of Lip-1 on neutrophilic asthma and ferroptosis by using the ovalbumin (OVA)/LPS-induced mouse model.. Lip-1 reversed the altered expression of ferroptotic regulators (glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11) and prostaglandin-endoperoxide synthase 2 (PTGS2)), attenuated lipid reactive oxygen species (lipid ROS) and ameliorated cell viability in HBE and BEAS-2B cells administered with LPS and IL-13. Moreover, Lip-1 treatment led to a marked reduction in the expression of IL-33, TSLP, IL-8, IL-6, and HMGB1 in the HBE and BEAS-2B cells. In the meantime, administration with Lip-1 markedly relieved OVA/LPS-induced neutrophilic asthma, as indicated by significant improvement in lung pathological changes, airway mucus secretion, inflammation, and ferroptosis.. This study provides data suggesting that Lip-1 alleviates neutrophilic asthma in vivo and in vitro through inhibiting ferroptosis, perhaps providing a new strategy for neutrophilic asthma treatment.

Topics: Animals; Asthma; Epithelial Cells; Ferroptosis; Inflammation; Interleukin-13; Lipopolysaccharides; Mice; Ovalbumin; Quinoxalines; Spiro Compounds

Follistatin-related protein 1 (FSTL1) is significantly associated with the asthma severity and outcome in humans and diverse mouse models of asthma. Previous studies have also suggested that FSTL1 could activate autophagy and NLRP3, thus playing as a causative agent in the asthma progression. However, mechanisms that regulate airway epithelial cell-specific FSTL1 expression and function in asthma are unknown. Here, we further evaluated the spatiotemporal relationships between the FSTL1 and asthma development through ovalbumin (OVA) -induced asthma models. Integrative analysis in asthmatics airway epithelium identifies microRNA (miR)-200b-3p as a novel upstream of FSTL1. Next, we collected airway biopsies, induced sputum, and blood samples isolated from asthmatics patients and the OVA-induced mouse model. We revealed that miR-200b-3p expression is downregulated in asthmatics airway epithelium, while its expression was negatively correlated with FSTL1. On this basis, the function and expression pattern analysis of miR-200b-3p were performed using miRNA-target prediction databases and long non-coding RNA (lncRNA) microarray assay. It is illustrated that miR-200b-3p, which is downregulated with pro-fibrotic stimulation of TGF-β1, could also be sponged by lncRNA PCAT19 and regulate FSTL1 expression in asthma progression. In vivo, miR-200b-3p overexpression in mice prevents OVA-induced airway remodeling and inflammation. Lastly, protective roles of miR-200b-3p are partly attributed to the direct and functional repression of FSTL1. Our findings suggest a crucial role for the miR-200b-3p/FSTL1 axis in regulating asthmatic's airway remodeling and inflammation phenotype.

Topics: Airway Remodeling; Animals; Asthma; Disease Models, Animal; Follistatin-Related Proteins; Humans; Inflammation; Mice; MicroRNAs; Ovalbumin; Phenotype; RNA, Long Noncoding

In respiratory diseases, the induction of allergic asthma has gradually aroused public concerns. Co-exposures of environmental risk factors such as nanoparticles and high humidity could play important roles in the development of allergic asthma. However, the relevant researches are still lacking and the involved mechanisms, especially metabolic changes, remain unclear. We took the lead in studying the combined induction effect and underlying mechanisms of carbon black nanoparticles (CB NPs) and high humidity on allergic asthma. In this work, murine models of allergic asthma were established with ovalbumin under the single and combined exposures of 15 μg/kg CB NPs and 90% relative humidity. The two risk factors, particularly their co-exposure, exhibited adjuvant effect on airway hyperresponsiveness, remodeling, and inflammation in Balb/c mice. Untargeted metabolomics identified the potential biomarkers in lung for asthma occurrence and for asthma exacerbation caused by CB NPs and high humidity. The significantly dysregulated metabolic pathways in asthmatic mice were proposed, and the disturbed metabolic pathways under the exposures of CB NPs and/or high humidity were mainly implicated in asthma symptoms. This work sheds light on the understanding for health risks of NP pollutions and high environmental humidity and contributes to useful biomarker identification and asthma control.

Topics: Animals; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humidity; Lung; Metabolome; Mice; Mice, Inbred BALB C; Nanoparticles; Ovalbumin; Soot

Asthma is a complex multifactorial chronic airway inflammatory disease with diverse phenotypes and levels of severity and is associated with significant health and economic burden. In a certain population of asthma patients, the symptoms cannot be well controlled with steroid. There has been long standing interest in the use of probiotics for treating allergic diseases. The purpose of this study is to investigate whether the combination of Lactobacillus rhamnosus GG (LGG) with prednisolone could reduce the dosage of glucocorticoid in controlling airway inflammation in a murine model for allergic asthma.. We used Der p 2-sensitized asthma model in female BALB/c mice. The animals were treated with 75 μl or 50 μl oral prednisolone or combination treatment of these two doses of oral prednisolone with LGG. Airway hyperresponsiveness, serum specific IgE/IgG1/IgG2a, infiltrating inflammatory cells in lung and cytokines were assessed.. Compared to 75 μl prednisolone, a lower dose of prednisolone with 50 μl was less satisfactory in suppressing airway hyperresponsives, serum IgE and IgG1, Th2 cytokines and inflammatory cytokines such as IL-6, IL-8 and IL-17 as well as infiltrating inflammatory cells. However, combination of 50 μl prednisolone and LGG decreased airway resistance and serum IgE and IgG1, inhibited the production of IL-4, IL-5, IL-6, IL-8, IL-13 and IL-17, upregulated serum IgG2a and enhanced Th1 immune response.. LGG may reduce the dosage of prednisolone and thus may be beneficial in the treatment of asthma.

Topics: Adrenal Cortex Hormones; Animals; Asthma; Cytokines; Disease Models, Animal; Female; Humans; Immunoglobulin E; Immunoglobulin G; Inflammation; Interleukin-17; Interleukin-6; Interleukin-8; Lacticaseibacillus rhamnosus; Mice; Mice, Inbred BALB C; Ovalbumin; Prednisolone

Alternative medicine, has become popular in asthmatic patients. We evaluated the immunomodulatory effects of SINA 1.2 therapy protocol derived from Persian medicine in an asthmatic mice model. Forty-two male BALB/c mice divided into six groups: one control (sham) and five sensitized groups (by parenteral injection of 20 μg ovalbumin in 100 μL normal saline plus 50 μL alum on days 1 and 14). Sensitized groups were as: untreated, budesonide (1 mg nebulized budesonide: 200 μg/puff every 5 min for 25 min), dry sauna (30 min, 37°C), oral oxymel (gavaged: 0.2 mL of the syrup plus 0.8 mL of water), and SINA protocol No.1.2 (oxymel followed by sauna) groups. Treatments were given for 10 days from day 23 to 33 then sacrificed. Significant gene expression reduction of interleukin(IL)-4, IL-5, and MUC5AC and increase of interferon(IFN)-γ and IFN-γ/IL-4 ratio and decreased perivascular and peribronchial inflammation, goblet cell hyperplasia, and subsequent mucus hypersecretion in SINA group were seen compared to untreated group. SINA lowered IL-5 and MUC5AC gene expression levels similar to the budesonide and acted better than budesonide in increasing IFN-γ gene expression up to normal level. Compared with the asthma group, sauna alone only affected MUC5AC and IFN-γ gene expressions and oxymel alone, only reduced IL-4 gene expression, perivascular and peribronchial inflammation, and mucus hypersecretion. It seems that SINA therapy alleviates asthma via immune modulation of pro-inflammatory cytokines and improvement of pathological changes in ovalbumin-induced asthma in mice, supporting the notion of innate healing power mentioned in Persian medicine literature.

Topics: Animals; Asthma; Budesonide; Humans; Inflammation; Interleukin-4; Interleukin-5; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Steam Bath

Ouabain, an inhibitor of Na+/K+-ATPase, is a type of endogenous hormone synthesized in the adrenal cortex and hypothalamus. Previous studies found that ouabain potently inhibited acute inflammatory reactions such as type 2 inflammation and regulated immunological processes. In this study, we aimed to investigate ouabain effect on allergic asthma.. BALB/c mice were submitted to chronic airway allergic inflammation induced by an ovalbumin (OVA) protocol. The animals were treated with ouabain or standard drug, budesonide. The following parameters were evaluated: cell migration, cytokine profile, IgE levels, lung histological modifications and MAPK activation.. At first, it was observed that ouabain reduced OVA-induced cell migration into the lung, observed by bronchoalveolar lavage fluid (BALF) cell counting and lung histological analysis (HE stain). Additionally, ouabain negatively modulated alarmins (IL-33 and TSLP), Th2 high cytokines levels (IL-1β and IL-4) and tissue remodeling markers such as TNF-α and TGF-β. Treatment with ouabain also reduced OVA-specific IgE titers in BALF and serum, respectively, when compared to the OVA group. Lung histological parameters, including collagen deposition and mucus production induced by OVA were also attenuated by ouabain treatment. Finally, our results showed that p38 mitogen-activated protein kinase (MAPK) signaling pathways were suppressed by ouabain in this model. All these parameters were reduced by budesonide, a steroidal anti-inflammatory standard drug.. These data together suggest that, in addition to its acute anti-inflammatory action, ouabain is also able to modulate allergic asthma.

Topics: Airway Remodeling; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Budesonide; Cytokines; Disease Models, Animal; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ouabain; Ovalbumin

Interleukin (IL)‑27 can inhibit the differentiation of Th2 cells and plays a role in the development of asthma. However, whether the therapeutic administration of IL‑27 in a mouse model of asthma can inhibit allergic responses remains a matter of debate. Additionally, the mechanisms through which IL‑27 ameliorates inflammatory responses in asthma are not yet fully understood. Thus, the aim of the present study was to examine the effects of IL‑27 on asthma using a mouse model and to elucidate the underlying mechanisms. For this purpose, mice received an intranasal administration of IL‑27 and the total and differential cell counts, levels of cytokines and type 1 regulatory T (Tr1) cells in the lungs were detected. The protein and mRNA levels of signal transducer and activator of transcription (STAT)1 and STAT3 were analyzed and airway remodeling was assessed. The results indicated that IL‑27 did not ameliorate airway inflammation, airway hyperresponsiveness, and airway remolding when administrated therapeutically. Preventatively, the administration of IL‑27 decreased the concentrations of Th2 cytokines and increased the number of Tr1 cells. The protein and mRNA levels of STAT1 and STAT3 were increased. Taken together, these findings demonstrate that the prophylactic administration of IL‑27 ameliorates asthma by alleviating the lung Th2 inflammatory environment through the restoration of both the STAT1 and STAT3 pathways. IL‑27 may thus prove to be useful as a novel agent for the prevention of asthma.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Interleukin-27; Interleukins; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; RNA, Messenger; Th2 Cells

Asthma is a lung disease that has influenced more than 350 million people worldwide. Airway smooth muscle (ASM) spasm leads to airway hyperresponsiveness (AHR) and bronchial obstruction, which are clinical manifestations of an asthma attack. Botulinum toxin (BTX) is a bacteria toxin that acts as muscle relaxant and may have therapeutic effects on AHR and asthma.. In this study, the effect of BTX on AHR and related gene expressions was evaluated.. An asthma mice model was developed which was treated with BTX in two ways: intranasally (IN) and via nebulization (N) (0.01, 0.1, and 1 U/mL and 10 U/mL, respectively) on days 25, 27 and 29. AHR was evaluated on days 24, 26, 28, and 30, and gene expressions were evaluated for TrkA, TrkB, M1-M5, α7nAChR, TNF-α, and extracellular signal-regulated kinase 2 (ERK2) proteins. For histopathology of the lungs, perivascular and peribronchial inflammation, production of mucus, and goblet cell hyperplasia were studied.. On day 24, treatment with BTX (for all doses) had no significant effect on AHR, but on days 26 and 28, AHR was decreased and this continued up to day 30 for all treated groups. Treatment with BTX had no significant effect on the gene expressions of TrkA, TrkB, M1-M5, α7nAChR, TNF-α, and ERK2 proteins, perivascular inflammation, peribronchial inflammation, hyperplasia of the goblet cell and production of mucus. Besides, mice administered with 10 mg/mL BTX perished. The BTX therapy controlled asthma attacks by decreasing AHR and relaxation of ASMs.. However, BTX had no significant effect on airway inflammation and production of mucus. While using BTX, it is necessary to prescribe safe doses in order to prevent adverse reactions.

Topics: alpha7 Nicotinic Acetylcholine Receptor; Animals; Asthma; Botulinum Toxins; Bronchi; Bronchial Hyperreactivity; Disease Models, Animal; Humans; Hyperplasia; Inflammation; Mice; Mice, Inbred BALB C; Myocytes, Smooth Muscle; Ovalbumin; Signal Transduction; Tumor Necrosis Factor-alpha

Loki zupa (LKZP) decoction, a traditional Uyghur medicine prescription, has been commonly used to treat numerous respiratory ailments in the Xinjiang region of western China, especially chronic airway inflammatory diseases such as allergic asthma. Due to its complex chemical composition, however, the mechanism of action of LKZP has yet to be fully elucidated.. Based on the balanced regulation theory of pro-inflammation and anti-inflammation, we tried to investigate the effectiveness of LKZP on asthma and its related protective mechanisms.. In this study, an experimental model of asthma was established using ovalbumin (OVA) in BALB/c mice to assess the effects of LKZP. The potential mechanism of LKZP anti allergic asthma were researched by the combination of in silico systems pharmacology and in vivo transcriptomics.. Our data revealed that LKZP exerted a therapeutic effect against OVA-induced asthma by reducing airway hyperresponsiveness (AHR), peribronchial inflammation, and mucus hypersecretion. Meanwhile, LKZP downregulated the expression of OVA-induced IgE, interleukin (IL)-4, IL-5, IL-13, and tumor necrosis factor (TNF)-α and concurrently promoted the expression of interferon (IFN)-γ in serum and bronchoalveolar lavage fluid (BALF). Systems pharmacology analysis identified 10 core bioactive ingredients and 26 hub targets of LKZP against asthma. Transcriptomic analysis confirmed 246 differentially expressed genes (DEGs) after LKZP treatment. These were mainly expressed in cytokine-cytokine receptor interactions and immune and inflammatory response-related signaling pathways. Additionally, the real-time quantitative PCR (qPCR) results for the nine selected DEGs matched those of the RNA-seq analysis. Nuclear factor (NF)-κB and hypoxia-inducible factor (HIF)-1 signaling pathways were identified as candidate targets involved in the action of LKZP on allergic asthma, which was highly consistent with the findings in silico. By qPCR, Western blot, and immunohistochemical analysis, it was verified that LKZP treatment dramatically inhibited the activation of NF-κB p65 and HIF-1α stimulated by OVA in asthmatic mice.. Taken together, our experimental data revealed that LKZP could be a candidate for the treatment of allergic asthma via NF-κB and HIF-1 signaling pathways.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Network Pharmacology; NF-kappa B; Ovalbumin; Transcriptome; Tumor Necrosis Factor-alpha

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; Vitamin E

Epidemiological and clinical studies have suggested that intake of n-3 polyunsaturated fatty acids (PUFA) reduces the incidence of allergic airway diseases and improves pulmonary function in patients with allergic asthma. However, the pharmacological targets of PUFA have not been elucidated upon. We investigated whether free fatty acid receptor 4 (FFA4, also known as GPR120) is a molecular target for beneficial PUFA in asthma therapy. In an ovalbumin (OVA)-induced allergic asthma model, compound A (a selective agonist of FFA4) was administrated before OVA sensitization or OVA challenge in FFA4 wild-type (WT) and knock-out (KO) mice. Compound A treatment of RBL-2H3 cells suppressed mast cell degranulation in vitro in a concentration-dependent manner. Administration of compound A suppressed in vivo allergic characteristics in bronchoalveolar lavage fluid (BALF) and lungs, such as inflammatory cytokine levels and eosinophil accumulation in BALF, inflammation and mucin secretion in the lungs. Compound A-induced suppression was not only observed in mice treated with compound A before OVA challenge, but in mice treated before OVA sensitization as well, implying that compound A acts on mast cells as well as dendritic cells. Furthermore, this suppression by compound A was only observed in FFA4-WT mice and was absent in FFA4-KO mice, implying that compound A action is mediated through FFA4. Activation of FFA4 may be a therapeutic target of PUFA in allergic asthma by suppressing the activation of dendritic cells and mast cells, suggesting that highly potent specific agonists of FFA4 could be a novel therapy for allergic asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Fatty Acids, Nonesterified; Humans; Lung; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin

Fisetin is isolated from many fruits and vegetables and has been confirmed to improve airway hyperresponsiveness in asthmatic mice. However, whether fisetin reduces inflammatory response and oxidative stress in bronchial epithelial cells is unclear. Here, BEAS-2B human bronchial epithelial cells were treated with various concentrations of fisetin and then stimulated with tumor necrosis factor-α (TNF-α) or TNF-α/interleukin-4. In addition, ovalbumin-sensitized mice were treated with fisetin to detect inflammatory mediators and oxidative stress expression. Fisetin significantly reduced the levels of inflammatory cytokines and chemokines in TNF-α-stimulated BEAS-2B cells. Fisetin also attenuated intercellular adhesion molecule-1 expression in TNF-α-stimulated BEAS-2B cells, suppressing THP-1 monocyte adhesion. Furthermore, fisetin significantly suppressed airway hyperresponsiveness in the lungs and decreased eosinophil numbers in the bronchoalveolar lavage fluid of asthmatic mice. Fisetin decreased cyclooxygenase-2 expression, promoted glutathione levels, and decreased malondialdehyde levels in the lungs of asthmatic mice. Our findings indicate that fisetin is a potential immunomodulator that can improve the pathological features of asthma by decreasing oxidative stress and inflammation.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Epithelial Cells; Flavonols; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Respiratory Hypersensitivity; Tumor Necrosis Factor-alpha

To probe into the ameliorative effect of Yanghe Decoction on pulmonary injury and immunologic derangement in asthmatic mice.. C57BL/6 mice were randomized into control (Con), Model, and Yanghe Decoction (YHF) groups, with 12 in each. The asthma model of adult female mice was induced by ovalbumin in the Model group, and the YHF group was treated by Yanghe Decoction on the basis of asthma modeling. The Con group received the same amount of normal saline. Inspiratory resistance (Ri), expiratory resistance (Re), lung compliance (CL), and maximal voluntary ventilation (MVV) were measured after modeling. Lung tissue was collected for the measurement of interleukin (IL)-4, IL-5, IL-6, IL-10, IL-13, and tumor necrosis factor-. The pulmonary function (PF) test revealed notably reduced Ri and Re as well as enhanced CL and MVV in asthmatic mice after the application of Yanghe Decoction. Yanghe Decoction dramatically ameliorated the pathological changes of lung tissue in asthmatic mice, as demonstrated by the staining results. ELISA results showed that Yanghe Decoction validly reduces lung tissue IL-4, IL-5, IL-6, IL-13, TNF-. Yanghe Decoction can effectively ameliorate the inflammatory reaction, immune cell disorder, and PF injury in ovalbumin-induced asthmatic mice.

Topics: Animals; Asthma; Drugs, Chinese Herbal; Female; Interleukin-10; Interleukin-13; Interleukin-5; Interleukin-6; Lung Injury; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Tumor Necrosis Factor-alpha

Recent studies have emphasized the role of endocrine-disrupting chemicals in asthma development, especially in eosinophilic asthma. However, the exact mechanism was unknown. Among all the endocrine-disrupting chemicals, mono-n-butyl phthalate (MnBP) was a chemical that was most frequently detected in human urine. Our study was performed with the aim of investigating the harmful effects of MnBP on airway epithelial cells (AECs), T cells, and eosinophils by using eosinophilic asthma mouse models. Mice that received OVA with MnBP had higher levels of airway hyperresponsiveness, total and eosinophil cell counts, as well as T cell proliferation and T helper 2 cytokine release than those which only received OVA. Moreover, MnBP contributed to directly enhancing the eosinophilic activation which was shown in. Long-term exposure MnBP activated AECs through the nuclear factor kappa B (NF-kB) pathway, decreased nuclear factor erythroid 2-related factor 2 (Nrf2) expression, and increased interleukin-33 expression. Additionally, MnBP can induce human eosinophil activation to release eosinophil extracellular traps (EETs). Taken together, our study suggested the roles of MnBP exposure increase the risk of asthma development and severity. Furthermore, vitamin E treatment (anti-inflammatory and antioxidant effects) can reduce MnBP-induced harmful effects through inhibiting EETs, restoring Nrf2, and suppressing the NF-kB pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Humans; Lung; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; NF-kappa B; Ovalbumin; Phthalic Acids

Th9- and regulatory T (Treg) cells exert pro- and anti-allergic activity, respectively. Mesenchymal stem cell (MSC)-related immunomodulatory impacts can be enhanced by inflammatory cytokines. Here, the modulatory effects of IFN-γ/TNF-α-induced MSCs on Th9- and Treg cell-related parameters were investigated using an asthma model.. Allergic asthma was induced in BALB/c mice using sensitized and challenging with ovalbumin (OVA). The asthmatic groups were treated intraperitoneally with PBS, MSCs, IFN-γ-induced MSCs, TNF-α-induced MSCs and 'IFN-γ + TNF-α'-induced MSCs before the challenge phase. The mice were sacrificed 24 h after challenge. The serum IL-9 and IL-35 levels, as well as gene expression of IL-9, PU.1, IL-35-EBI3, and FOXP3 in the lung tissues were assessed using ELISA and real time-PCR, respectively.. The differences of Th9 and Treg-related parameters were not significant between untreated asthmatic mice and those treated with non-induced MSCs. In comparison with untreated asthmatic group, treatment with IFN-γ-induced MSCs significantly reduced serum IL-9 levels, reduced lung expression of IL-9 and PU.1, while increasing serum IL-35 levels as well as lung expression of FOXP3; treatment with TNF-α-induced MSCs significantly reduced serum IL-9 levels as well as lung expression of IL-9, and treatment with 'IFN-γ + TNF-α'-induced MSCs, significantly modulated all investigated Th9 and Treg-related parameters. In comparison to mice treated with non-induced MSCs, serum IL-9 levels were remarkably decreased in mice treated with IFN-γ-induced and 'IFN-γ + TNF-α'-induced MSCs.. IFN-γ-and 'IFN-γ + TNF-α' treated MSCs exerted almost comparable impacts, but were more efficient than TNF-α-exposed MSCs. Thus, IFN-γ alone can be sufficient to promote immunomodulatory effects of MSCs.

Topics: Animals; Anti-Allergic Agents; Asthma; Cytokines; Disease Models, Animal; Forkhead Transcription Factors; Interferon-gamma; Interleukin-9; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Tumor Necrosis Factor-alpha

Topics: Animals; Anti-Inflammatory Agents; Asthma; Cyclooxygenase 2; Disease Models, Animal; Fagaceae; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Plant Extracts; Seeds

Lycopene as the main carotenoid from tomatoes is known to have beneficial effects on various inflammatory diseases. In mice, lycopene ameliorates asthma symptoms and in human asthmatic patients serum lycopene levels are reduced. To further investigate the immunomodulatory effect of lycopene, first, we used a ragweed pollen extract (RWE)-induced asthma model in mice. In a second approach, we established a RWE-induced asthma model in gerbils, because of a more human-like carotenoid absorption in these animals. In RWE-sensitized/RWE-challenged gerbils (C+) following a basal diet, mainly the number of eosinophils in the broncho-alveolar lavage (BAL) significantly increased, comparable to RWE-sensitized/PBS-challenged gerbils (C-). In RWE-sensitized/PBS-challenged gerbils with lycopene-supplementation (L-), an elevated number of mainly neutrophils, in addition to eosinophils, was detected compared to C-, whereas in RWE-sensitized/RWE-challenged animals with lycopene-supplementation (L+), mainly increased neutrophil numbers in BAL were detected compared to C+. Furthermore, using LC-MS, we determined an array of eicosanoids/docosanoids in the lungs and observed that 5-, 8-lipoxygenase (LOX) and cyclooxygenase (COX) pathways were significantly increased after intranasal RWE-challenge in sensitized mice and just by tendency in gerbils. In PBS- and RWE-challenged animals, lycopene-supplementation significantly raised COX-pathway metabolites. In conclusion, we found that lycopene-supplementation resulted in an increased inflammatory influx of neutrophils in combination with increased COX-pathways metabolites. This pro-inflammatory, pro-neutrophil activity induced by lycopene might be an important shift from allergic asthma towards an inflammatory symptomatic asthma type, though with the potential for resolution.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Humans; Inflammation; Lycopene; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin

Topics: Animals; Asthma; Cytokines; Dexamethasone; Disease Models, Animal; Eosine Yellowish-(YS); Glutathione Peroxidase; Hematoxylin; Interleukin-13; Interleukin-4; Interleukin-5; Luteolin; Malondialdehyde; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; NF-E2-Related Factor 2; NF-kappa B; NF-KappaB Inhibitor alpha; Ovalbumin; Reactive Oxygen Species; Superoxide Dismutase

Ion channels are potentially exploitable as pharmacological targets to treat asthma. This study evaluated the role of KCa3.1 channels, encoded by

Topics: Animals; Asthma; Bronchi; Disease Models, Animal; Gene Expression; Mice; Mice, Inbred BALB C; Ovalbumin; Trachea

Autoimmune, allergic, and respiratory inflammatory diseases are some of the most important health issues worldwide. Disorders of the gut microbiota have been associated with the induction of allergic and inflammatory diseases, and probiotics are being tested for disease prevention. We examined functional Lactiplantibacillus plantarum RGU (Lp-1) to mice with ovalbumin (OVA)-induced asthma model to elucidate the inhibitory effect on pathological progression in asthma model. Prior to the experiments, the intestinal lactic acid bacteria were reduced by administering multiple antibiotics (MAB) to evaluate the administration effect of lactic acid bacteria. Mice were administered with Lp-1 or comparative control lactic acid bacteria in each group. After that, OVA-induced asthma was induced, and cytokine gene expression and histological findings were compared. Exacerbation of lung lesions was confirmed in the MAB group. The Lp-1 group mice had alleviated lung lesions with a decrease in IL-1β, IL-13, IL-17 and an increase in IL-10 of the splenocytes and bronchial lymph nodes compared with the MAB group, but not in the other groups. In OVA-induced asthma, administration of specific Lactiplantibacillus was confirmed to induce anti-inflammatory cytokines.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Eosinophils; Female; Flavanones; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Respiratory Hypersensitivity; Sophora

High levels of allergen exposure increase the prevalence of asthma in developed countries. The asthmatic type 2 T-helper (Th2) response is characterized with high eosinophil infiltration, elevated Th2 cytokines, and immunoglobulin (Ig) E secretion resulting in local or systemic inflammation. However, the treatment with palliative Th2 inhibitor drugs cannot completely control asthma and that is why the development of novel approaches is still important. Based on type 1 T-helper (Th1) and Th2 immune homeostasis, the enhanced Th1 immune response has high potential to alleviate Th2 immune response. Thus, we aimed to overexpress single chain IL-12 (scIL-12) through recombinant adeno-associated virus (rAAV) vector (as rAAV-IL-12) and test the efficacy in an ovalbumin (OVA)-induced asthmatic murine model. We firstly demonstrated the bioactivity of exogenous scIL-12. The expression of exogenous scIL-12 was also detected in the lungs of rAAV-IL-12 transduced mice. The data demonstrated that overexpression of exogenous scIL-12 significantly suppressed total number of cells and eosinophil infiltration, as well as the mucus secretion in rAAV-IL-12-treated mice. The decreased OVA-specific IgE in bronchoalveolar lavage fluid and gene expression of Th2-cytokines or C-C motif ligand (CCL) 11 (also eotaxin-1) in lung were observed. In addition, the production of cytokines in the supernatants of OVA-stimulated splenocytes were suppressed with rAAV-IL-12 treatment. Thus, scIL-12 expression by rAAV vector was able to modulate Th2 activity and has the potential to be developed as a feasible strategy in modulating allergic diseases.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Cytokines; Dependovirus; Gene Expression; Immunoglobulin E; Interleukin-12; Ligands; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Th2 Cells

Clinical observations have shown the risk of cardiovascular disease during asthmatic changes. Whether and how asthma causes heart failure is the subject of debate. Here, we aimed to investigate the possibility of cardiomyocyte mitophagy in a rat model of asthma. Twelve mature Wistar rats were randomly allocated into the Control and Asthmatic rats (n = 6). To induce asthma, ovalbumin was injected intraperitoneally on days 1 and 8 and procedure followed by nebulization from days 14 to 32. After 2 weeks, we performed the pathological examination of both lungs and heart using Hematoxylin-Eosin staining. Real-time PCR analysis was used to measure the expression of mitophagic factors, such as Optineurin, Pink1, and mitofusin 1 and 2. Typical changes like increased inter-alveolar septa thickness and interstitial pneumonia were evident in asthmatic lungs. In cardiac tissue, slight inflammatory response, and hydropic degeneration with an eosinophilic appearance were detected in the cytoplasm of cardiomyocytes. Real-time PCR analysis showed mitophagic response in pulmonary and cardiac tissues via upregulation of mitophagy-related genes like Optineurin and Pink-1 in asthmatic lungs and hearts compared to the control group (p < 0.05). Likewise, asthmatic changes increased the expression of genes associated with mitochondrial fusion in the lungs and heart. The expression of mitofusin1 and 2 was significantly increased following inflammatory response in pulmonary and cardiac tissues (p < 0.05). Our findings showed the expression of certain factors related to mitophagy during chronic asthmatic conditions. The findings open a new avenue in the understanding of cardiomyocyte injury during asthma.

Topics: Animals; Asthma; Lung; Mitophagy; Myocytes, Cardiac; Ovalbumin; Rats; Rats, Wistar

Pinelliae Rhizoma Praeparatum (PRP) is a traditional processed product of Pinellia ternata (Thunb.) Berit., which mainly used for treating cold asthma (CA). However, the mechanism of action of PRP for treating CA have not been fully elucidated.. To investigate the core active constituents and the pharmacological mechanism of PRP against CA.. Ovalbumin (OVA) and cold water-induced cold asthma model were established in male mice. The effects of water extract from PRP were evaluated by general morphological observation, expectorant activity, airway hyperresponsiveness, mucus hypersecretion, inflammatory cytokines, etc. Additionally, the mRNA and protein expression of mucin 5AC (MUC5AC) and aquaporin 5 (AQP5) in vivo and in vitro were detected by immunohistochemistry (IHC), qRT-PCR, and western blotting. The mechanisms of action were investigated through network pharmacology and transcriptomic, and validated through western blotting and molecular docking.. PRP exhibited a favorable expectorant activity, and significantly reduced the airway inflammation, mucus secretion, and hyperresponsiveness in cold asthma model. It also reduced the levels of IL-4, IL-5, IL-8, and IL-13 in bronchoalveolar lavage fluid (BALF) and IL-4 and total IgE in serum, while obviously increased the levels of IL-10 and IFN-γ in serum for asthmatic mice. Meanwhile, PRP also attenuated the pathological changes and mucus production in cold asthmatic mice. Moreover, the downregulation of MUC5AC and upregulation of AQP 5 were detected by western blotting and qRT-PCR after administration with PRP both in vivo and in vitro. PRP expectedly inhibited the protein expression of PKC-α, SRC, p-EGFR, p-ERK1/2, p-JNK, p-p38, p-PI3K, and p-Akt levels in vivo.. These combined data showed that PRP suppressed the allergic airway inflammation of CA by regulating the balance of Th1 and Th2 cytokines and the possible involvement of the PKC/EGFR/MAPK/PI3K-Akt signaling pathway. Pentadecanoic acid, licochalcone A, β-sitosterol, etc. were considered as main active ingredients of PRP against CA. This study provides a novel perspective of the classical herbal processed product PRP in the treatment of CA.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; ErbB Receptors; Expectorants; Inflammation; Interleukin-4; Lung; Male; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Molecular Docking Simulation; Mucus; Ovalbumin; Phosphatidylinositol 3-Kinases; Pinellia; Proto-Oncogene Proteins c-akt; Signal Transduction; Water

Ziziphus mauritiana Lam leaves were utilized in treating asthma, diabetes, inflammation, and hepatic diseases in Indian traditional medicine. The leaves were used as an edible vegetables in rural parts of India.. The aim is to prove the anti-inflammatory activity of Ziziphus mauritiana Lam leaves against LPS-stimulated RAW 264.7 macrophages and OVA-induced airway inflammation in mice through its attenuation mechanism in the NFκB signalling pathway.. Terpenoids present in MEZ were quantified using U(H)PLC analysis. MEZ at 50 and 100 μg/mL were tested against LPS stimulated RAW 264.7 macrophages. The concentration of NO, ROS, and cytokines was quantified from the cell culture supernatants. OVA-induced asthma in mice was adopted for screening airway inflammation. MEZ at 250 and 500 mg/kg was tested for airway hyperresponsiveness, leukocyte counting, pro-inflammatory cytokines (IL-4, IL-5, IL-13 and TNF-α), lung histopathology, and various inflammatory gene expressions in lungs for NFκB signalling pathway in asthma.. Terpenoids like betulin, betulinic acid, oleanolic acid, and ursolic acid were quantified from U(H)PLC analysis. MEZ at higher doses reduced the NO, ROS, and pro-inflammatory cytokines in LPS stimulated RAW 264.7 macrophages. MEZ at 500 mg/kg significantly reduced AHR and also decreased total and differential leukocytes. MEZ also reduced the expressions of ICAM, VCAM, and Muc5C genes. Histopathological analysis revealed MEZ significantly reduced the leukocyte infiltration and mucus hypersecretion in the lungs. MEZ suppressed lung inflammation by inhibition of p65 mediated IκB-α translocation in the NFκB signalling pathway.. From these findings, MEZ significantly reduced airway inflammation by inhibiting NFκB mediated inflammatory pathway. Hence, this study proved that Ziziphus mauritiana Lam has anti-asthmatic potential in Indian traditional medicine.

Topics: Animals; Asthma; Cytokines; Inflammation; Lipopolysaccharides; Macrophages; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Reactive Oxygen Species; Terpenes; Ziziphus

To clarify the detailed molecular mechanisms underlying the development of asthma, we assessed the potential immune effects of prenatal osteoprotegerin (OPG) inhibition in the pathogenesis of asthma. The effects of OPG deficiency on the development of asthma were evaluated using an ovalbumin-induced asthma model in OPG knockout mice. Histological analysis demonstrated that OPG was mainly detected in airway epithelial cells in wild type mice. After ovalbumin sensitization and challenge, accumulation of inflammatory cells, gene expression of T helper 2-related cytokines and mucus hypersecretion in lung tissues were inhibited by OPG deficiency. Importantly, the serum level of IgE was not increased in OPG KO mice after ovalbumin sensitization and challenge. Based on these findings, OPG knockout mice were protected against methacholine-induced airway hyperresponsiveness. OPG expression is thought to be essential for induction of the allergic immune response in asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Mice, Knockout; Osteoprotegerin; Ovalbumin

To investigate the effect of IL-27 on Th9 differentiation and Th1/Th2 balance.. C57BL/6 (B6) mice were treated with ovalbumin to establish an allergic asthma (AA) model and subjected to IL-27 overexpression (OV) and empty vector (EV). Hematoxylin-eosin (HE) staining was performed to observe lung tissue inflammation. Flow cytometry was carried out to evaluate the percentage of Th9, Th1, and Th2 cells. The expression of IL-27, IL-27R, IL-9, T-bet, IFN-γ, and IgE was evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). Western blot was conducted to observe the expression of pSTAT-1 and pSTAT-3.. Compared with the Model group, the number of Th1 cells in the Model + OV group increased significantly (. IL-27 OV inhibits Th9 differentiation and regulates the imbalance of Th1/Th2, thereby alleviating inflammatory response in AA. The findings suggest that IL-27 OV may be a potential strategy for clinical treatment of AA.

Topics: Animals; Asthma; Disease Models, Animal; Eosine Yellowish-(YS); Hematoxylin; Immunoglobulin E; Interleukin-27; Interleukin-9; Interleukins; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Th1 Cells; Th2 Cells

Allergic asthma is a chronic disease and medical treatment often fails to fully control the disease in the long term, leading to a great need for new therapeutic approaches. Immunoproteasome inhibition impairs T helper cell function and is effective in many (auto-) inflammatory settings but its effect on allergic airway inflammation is unknown.. Immunoproteasome expression was analyzed in. These results show the importance of the immunoproteasome in Th2 cells and airway inflammation. Our data provides first insight into the potential of using immunoproteasome inhibition to target the aberrant Th2 response, e.g. in allergic airway inflammation.

Topics: Animals; Asthma; Disease Models, Animal; Inflammation; Mice; Ovalbumin; Th17 Cells; Th2 Cells

Asthma is a major cause of morbidity and mortality in humans. The mechanisms of asthma are still not fully understood. Leukocyte-specific protein-1 (LSP-1) regulates neutrophil migration during acute lung inflammation. However, its role in asthma remains unknown.. Light and electron microscopic immunocytochemistry and Western blotting showed that, compared with normal healthy lungs, the levels of LSP1 were increased in lungs of OVA-asthmatic mice. Compared to Lsp1. These data show that LSP1 deficiency reduces airway hyper-responsiveness and lung inflammation, including leukocyte recruitment and cytokine expression, in a mouse model of asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Respiratory Hypersensitivity

In respiratory diseases, the induction of allergic asthma is one of the hottest issues of international concern. The adjuvant effect of air pollutants including nanoparticles (NPs) has be pointed out to facilitate the occurrence and development of allergic asthma. This work studied the development of allergic asthma upon exposures of carbon black nanoparticles (CB NPs, 30-50 nm) and/or high environmental humidity (90% relative humidity). The mechanisms involved were investigated from perspectives of the activation of oxidative stress and transient receptor potential vanilloid 1 (TRPV1) pathways and the alteration in intestinal microbiota. Both high humidity and CB NPs aggravated the airway hyperreactivity, remodeling, and inflammation in Balb/c mice sensitized by ovalbumin. The co-exposure of these two risk factors exhibited adjuvant effect on the development of asthma likely through activating oxidative stress pathway and TRPV1 pathway and then facilitating type I hypersensitivity. Additionally, exposures of high humidity and/or CB NPs reduced the richness of intestinal microbes, altered microbial community composition, and weakened corresponding biological functions, which may interact with the development of asthma. The findings will add new toxicological knowledge to the health risk assessment and management of co-exposures of NPs and other risk factors in the environment.

Topics: Animals; Asthma; Disease Models, Animal; Gastrointestinal Microbiome; Humidity; Lung; Mice; Mice, Inbred BALB C; Nanoparticles; Ovalbumin; Soot

Renifolin F is a prenylated chalcone isolated from

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Chalcone; Cytokines; Disease Models, Animal; Hypersensitivity; Immunity, Innate; Inflammation; Lung; Lymphocytes; Mice; MicroRNAs; Ovalbumin

Maternal improper nutrition has been reported to trigger respiratory disorders in offspring. Here, we characterized the effects of high-fat environment in the fetal period on mice and human cord blood CD4

Topics: Animals; Asthma; CD4-Positive T-Lymphocytes; Cell Differentiation; Cytokines; Diet, High-Fat; Disease Models, Animal; Hypersensitivity; Mice; Ovalbumin

Despite the substantial amount of efforts made to reduce morbidity and improve respiratory management, asthma control remained a major challenge for severe patients. Plant isoflavones, one of the most estrogenic compounds, are considered a potential alternative therapy for asthma. Iristectorigenin A, a naturally occurring isoflavone, is extracted from a variety of medical plants and its biological activity has not been reported previously.. In present study, we aim to reveal the potential therapeutic role of Iristectorigenin A against acute asthmatic mice.. We established ovalbumin (OVA) induced asthmatic murine model and orally administrated Iristectorigenin A at concentration of 5 and 10 mg/kg and dexamethasone as a positive control substance.. Asthmatic murine model was established with OVA sensitization and challenge. Lung function was assessed with FinePoint Ventilation system recording lung resistance (RI) and lung compliance (Cydn). White cells were sorted and counted in BALF. Histopathological assessment was conducted by H&E, PAS, and Masson's trichrome staining on paraffin embedded lung tissues. BALF content of IL-4, IL-5, IL-33, IL-13, INF-γ, IL-9 and serum IgE, IgG1 were measured using ELISA kit. Expression levels of mRNAs associated with inflammatory cytokines and goblet cell metaplasia were evaluated via quantitative RT-PCR. Protein expression levels of FOXA3, MUC5AC, SPDEF were estimated by immunohistochemistry on lung tissue, while NOTCH1 and NOTCH2 expressions were evaluated by western blotting analysis.. Iristectorigenin A resulted in improved airway hyperresponsiveness (AHR) mirrored by decreased RI and increased Cydn. With Iristectorigenin A, we also observed reduced number of BALF leukocytes, improved inflammatory cell infiltration in lung tissue, decreased content of BALF IL-4, IL-5, IL-33, but not IL-13, INF-γ, IL-9, and their mRNA levels, along with decreased levels of OVA-specific IgE, IgG1 in asthmatic mice. Additionally, Iristectorigenin A exhibited significant therapeutic potential on attenuating mucus production reflected by mitigated FOXA3 and MUC5AC immunostaining on the airway epithelium, as well as decreased mRNAs associated with goblet cell metaplasia. At last, a decrease in elevated expression level of NOTCH2, but not NOTCH1, in asthmatic mice lung tissue was observed by western blotting analysis.. Our study provides strong evidence that Iristectorigenin A can be potential therapeutic agent ameliorating airway inflammation and mucus hypersecretion in allergic asthma. This is a first research reported the potential of Iristectorigenin A as an alternative therapeutic agent.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Immunoglobulin E; Immunoglobulin G; Inflammation; Interleukin-33; Interleukin-4; Interleukin-5; Interleukin-9; Lung; Metaplasia; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Phenotype

Dioscorea nipponica Makino (DNM) is a traditional herb with multiple medicinal functions. This study is aimed at exploring the therapeutic effects of DNM on asthma and the underlying mechanisms involving RKIP-mediated MAPK signaling pathway.. An ovalbumin-induced asthma model was established in mice, which was further administrated with DNM and/or locostatin (RKIP inhibitor). ELISA was performed to detect the serum titers of OVA-IgE and OVA-IgG1, bronchoalveolar lavage fluid (BALF) levels of inflammation-related biomarkers, and tissue levels of oxidative stress-related biomarkers. The expression of RKIP was measured by quantitative real-time PCR, Western blot, immunohistochemistry, and immunofluorescence. HE staining was used to observe the pathological morphology of lung tissues. The protein expression of MAPK pathway-related proteins was detected by Western blot.. DNM relieves asthma via blocking the Raf-1/MEK/MAPK/ERK pathway that mediated by RKIP upregulation.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dioscorea; Disease Models, Animal; Immunoglobulin E; Immunoglobulin G; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinase Kinases; Ovalbumin; Phosphatidylethanolamine Binding Protein; Plant Extracts; Proto-Oncogene Proteins c-raf

Type 2-high asthma is a chronic inflammatory disease of the airways which is increasingly prevalent in countries where helminth parasite infections are rare, and characterized by T helper 2 (Th2)-dependent accumulation of eosinophils in the lungs. Regulatory cytokines such as TGF-β can restrain inflammatory reactions, dampen allergic Th2 responses, and control eosinophil activation. The murine helminth parasite Heligmosomoides polygyrus releases a TGF-β mimic (Hp-TGM) that replicates the biological and functional properties of TGF-β despite bearing no structural similarity to the mammalian protein. Here, we investigated if Hp-TGM could alleviate allergic airway inflammation in mice exposed to Alternaria alternata allergen, house dust mite (HDM) extract or alum-adjuvanted ovalbumin protein (OVA). Intranasal administration of Hp-TGM during Alternaria exposure sharply reduced airway and lung tissue eosinophilia along with bronchoalveolar lavage fluid IL-5 and lung IL-33 cytokine levels at 24 h. The protective effect of Hp-TGM on airway eosinophilia was also obtained in the longer T-cell mediated models of HDM or OVA sensitisation with significant inhibition of eotaxin-1, IL-4 and IL-13 responses depending on the model and time-point. Hp-TGM was also protective when administered parenterally either when given at the time of allergic sensitisation or during airway allergen challenge. This project has taken the first steps in identifying the role of Hp-TGM in allergic asthma and highlighted its ability to control lung inflammation and allergic pathology. Future research will investigate the mode of action of Hp-TGM against airway allergic eosinophilia, and further explore its potential to be developed as a biotherapeutic in allergic asthma.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Chemokine CCL11; Cytokines; Eosinophilia; Helminths; Interleukin-13; Interleukin-33; Interleukin-4; Interleukin-5; Lung; Mammals; Mice; Mice, Inbred BALB C; Ovalbumin; Transforming Growth Factor beta

Aeroallergens are common inducers of asthma in humans and are widely used in experimental research to generate animal models of this disease. In this chapter, we describe four mouse models of aeroallergen-induced asthma. These models differ in type and number of allergens used, route and duration of allergen exposure, and utilization of an adjuvant, representing different mechanistic variants of asthma. In addition, we describe several basic methods that are commonly used in mechanistic studies of asthma in mice. These methods include tracheotomy and bronchoalveolar lavage, cytospin and morphologic analysis of bronchoalveolar lavage cells, and lung harvest and digestion for generation of single-cell suspension.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Asthma has been the most prevalent chronic respiratory disease (Mensah et al. J Allergy Clin Immunol 142:744-748, 2018). To explore pathogenic mechanism or new treatments of asthma, mice have been utilized to model the disease. Eosinophilic airway inflammation, allergen specific-IgE, and airway hyperresponsiveness have been characteristic features of allergic asthma (Drake et al. Pulm Ther 5:103-115, 2019). In mouse models, airway hyperresponsiveness to inhaled broncho-constrictor agents such as methacholine chloride (MCh) has been a key disease marker (Alessandrini et al. Front Immunol 11:575936, 2020). A variety of systems to assess airway reactivity in mice are currently available. Here, three distinct systems are described as these have been used in many publications. In the first system, an invasive system in which mice are anesthetized and intubated followed by mechanical ventilation, lung resistance (R), dynamic compliance (C), and other respiratory parameters with MCh challenge are measured. In the second system, a noninvasive system equipped with a chamber in which mice can move freely and spontaneously breathe, changes in airways with MCh challenge are measured as enhanced pause (Penh) values. In the third system, in vitro airway smooth muscle (ASM) reactivity is monitored in an extracted mouse tracheal duct with a cholinergic agonist challenge or electrical stimulation. Each of these systems has unique features, benefits, or disadvantages.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Disease Models, Animal; Eosinophilia; Immunoglobulin E; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Respiration Disorders; Respiratory Hypersensitivity

Allergic asthma is an airway inflammatory disease dominated by type 2 immune responses and there is currently no curative therapy for asthma. CD5-like antigen (CD5L) has been known to be involved in a variety of inflammatory diseases. However, the role of CD5L in allergic asthma remains unclear. In the present study, mice were treated with recombinant CD5L (rCD5L) during house dust mite (HDM) and ovalbumin (OVA) challenge to determine the role of CD5L in allergic asthma, and the underlying mechanism was further explored. Compared with PBS group, serum CD5L levels of asthmatic mice were significantly decreased, and the levels of CD5L in lung tissues and bronchoalveolar lavage fluid (BALF) were significantly increased. CD5L reduced airway inflammation and Th2 immune responses in asthmatic mice. CD5L exerted its anti-inflammatory function by increasing CD11c

Topics: Animals; Apoptosis Regulatory Proteins; Asthma; Bronchoalveolar Lavage Fluid; CD11c Antigen; Disease Models, Animal; Histone Deacetylase 2; Inflammasomes; Inflammation; Lung; Macrophages, Alveolar; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Receptors, Scavenger

Subgroups of patients with severe asthma showing marked increases in sputum eosinophils and/or neutrophils are insensitive to corticosteroids. Previous reports have shown that exogenous administration of an anti-inflammatory cytokine, interleukin (IL)-10 negatively regulated both eosinophilic and neutrophilic migration into tissues. The objective of this study was to elucidate whether intratracheal IL-10 administration suppresses asthmatic responses in a steroid-insensitive model of mice. Ovalbumin (OVA)-sensitized BALB/c mice were intratracheally challenged with OVA at 500 µg/animal four times. Dexamethasone (1 mg/kg, intraperitoneal) or IL-10 (25 ng/mouse, intratracheal) was administered during the multiple challenges. The number of leukocytes, expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and IL-10 receptor in the lung, and the development of airway remodeling and hyperresponsiveness were evaluated after the fourth challenge. Consistent with our previous study, dexamethasone hardly suppressed the development of airway remodeling and hyperresponsiveness. Although intratracheal IL-10 administration did not affect the development of airway remodeling, the infiltration of eosinophils and neutrophils, and the development of airway hyperresponsiveness were significantly inhibited. Moreover, IL-10 administration significantly decreased the numbers of ICAM-1

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dexamethasone; Disease Models, Animal; Endothelial Cells; Eosinophils; Intercellular Adhesion Molecule-1; Interleukin-10; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Interleukin-10; Respiratory Hypersensitivity; Steroids; Vascular Cell Adhesion Molecule-1

Asthma is the most prevalent chronic respiratory disease worldwide. There is currently no cure, and it remains an important cause of morbidity and mortality. Here we report that lung-specific loss of function of the transcription factor Miz1 (c-Myc-interacting zinc finger protein-1) upregulates the pro-T-helper cell type 1 cytokine IL-12. Upregulation of IL-12 in turn stimulates a Th1 response, thereby counteracting T-helper cell type 2 response and preventing the allergic response in mouse models of house dust mite- and OVA (ovalbumin)-induced asthma. Using transgenic mice expressing Cre under a cell-specific promoter, we demonstrate that Miz1 acts in lung epithelial cells and dendritic cells in asthma. Chromatin immunoprecipitation coupled with high-throughput DNA sequencing or quantitative PCR reveals the binding of Miz1 on the

Topics: Animals; Asthma; Disease Models, Animal; Interleukin-12; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Protein Inhibitors of Activated STAT; Pyroglyphidae; Th1 Cells; Th2 Cells; Ubiquitin-Protein Ligases

It is challenging to overcome difficult-to-treat asthma, and cell-based therapies are attracting increasing interest. We assessed the effects of mesenchymal stem cell (MSC) treatments using a murine model of chronic ovalbumin (OVA)-challenged asthma. We developed a murine model of chronic allergic asthma using OVA sensitization and challenge. Human adipose-derived MSCs (hADSCs) or human bone marrow-derived MSCs (hBMSCs) were administered. We measured the levels of resistin-like molecule-β (RELM-β). We also measured RELM-β in asthma patients and normal controls. OVA-challenged mice exhibited increased airway hyper-responsiveness, inflammation, and remodeling. hBMSC treatment remarkably decreased airway hyper-responsiveness but hADSC treatment did not. Both MSCs alleviated airway inflammation, but hBMSCs tended to have a more significant effect. hBMSC treatment reduced Th2-cytokine levels but hADSC treatment did not. Both treatments reduced airway remodeling. The RELM-β level decreased in the OVA-challenged control group, but increased in both treatment groups. We found that the serum level of RELM-β was lower in asthma patients than controls. MSC treatments alleviated the airway inflammation, hyper-responsiveness, and remodeling associated with chronic asthma. hBMSCs were more effective than hADSCs. The RELM-β levels increased in both treatment groups; the RELM-β level may serve as a biomarker of MSC treatment efficacy.

Topics: Adipose Tissue; Airway Remodeling; Animals; Asthma; Bone Marrow; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humans; Inflammation; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity

Endocannabinoids are biologically active cannabinoid-related substances endogenously synthesized in many mammalian tissues. Mainly two enzymes carry out their degradation; Fatty Acid Amide Hydrolase (FAAH) and Monoacylglycerol Lipase (MAGL). Endocannabinoids are shown to affect the modulation of inflammatory processes and airway responsiveness. In the present study, we investigated the effects of FAAH and MAGL inhibitor treatments in experimental allergic airway inflammation in guinea pigs.. Guinea pigs were sensitized and challenged by ovalbumin to induce an allergic asthma model. Then, the effects of FAAH inhibitor URB597, MAGL inhibitor JZL184, and dual (FAAH/MAGL) inhibitor JZL195 on airway inflammation and hyperreactivity were evaluated.. Ovalbumin challenge increased airway reactivity, IgE in serum, IL-4, and IL-13, and the percentage of eosinophils in bronchoalveolar lavage (BAL). In addition, inhibition of FAAH or MAGL enzymes leads to an increase in endocannabinoid levels. The selective inhibition of the FAAH enzyme prevented inflammation indicators such as cytokine production and inflammatory cell infiltration but had a negligible effect on airway hyperreactivity. However, the inhibition of the MAGL enzyme or dual inhibition of both FAAH and MAGL enzymes tent to moderate both pulmonary inflammation and airway hyperreactivity.. We have previously demonstrated that modulation of endocannabinoid levels in the airways by FAAH or MAGL inhibition can be useful in preventing acute lung inflammation. The results of the present study further suggest that FAAH and MAGL inhibitor treatment can also be a promising strategy for bronchial hyperreactivity and airway inflammation in allergic asthma.

Topics: Amidohydrolases; Animals; Asthma; Endocannabinoids; Enzyme Inhibitors; Guinea Pigs; Inflammation; Mammals; Monoacylglycerol Lipases; Ovalbumin

To investigate the effects of β-sitosterol (B-SIT) and the underlying mechanisms of action in an ovalbumin-induced rat model of asthma.. The pathological and morphological changes in lung and tracheal tissues were observed by H&E, PAS, and Masson's staining. The levels of IgE, TNF-. B-SIT improved the injury in OVA-induced pathology, decreased the levels of inflammatory factors of IgE, TNF-. B-SIT improved symptoms in a rat model of asthma likely

Topics: Animals; Asthma; Dendritic Cells; Interleukin-10; Interleukin-13; Interleukin-5; Interleukin-6; Ovalbumin; Rats; Sitosterols; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Eosinophils; Gynostemma; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Th2 Cells

Asthma is a chronic airway disorder with a hallmark feature of airflow obstruction that associated with the remodeling and inflammation in the airway wall. Effective therapy for controlling both remodeling and inflammation is still urgently needed. Leonuride is the main pharmacological component identified from Bu-Shen-Yi-Qi-Tang (BSYQT) which has been traditionally used in treatment of lung diseases. However, no pharmacological effects of leonuride in asthma were reported.. Here we aimed to investigated whether leonuride provided a therapeutic efficacy in reversing asthma airway remodeling and inflammation and uncover the underlying mechanisms.. Mouse models of chronic asthma were developed with ovalbumin (OVA) exposure for 8 weeks. Respiratory mechanics, lung histopathology and asthma-related cytokines were examined. Lung tissues were analyzed using RNA sequencing to reveal the transcriptional profiling changes.. After oral administration with leonuride (15 mg/kg or 30 mg/kg), mice exhibited a lower airway hyperresponsiveness in comparison to asthmatic mice. Leonuride suppressed airway inflammation evidenced by the significant reductions in accumulation of inflammatory cells around bronchi and vessels, leukocyte population counts and the abundance of type 2 inflammatory mediators (OVA specific IgE, IL-4, IL-5 and IL-13) in bronchoalveolar lavage fluid (BALF). On the other hand, leonuride slowed down the process of active remodeling as demonstrated by weaker goblet cell metaplasia and subepithelial fibrosis in lung histopathology and lower transforming growth factor (TGF)-β1 levels in serum and BALF in comparison to mice treated with OVA only. Furthermore, we uncovered transcriptional profiling alternations in lung tissue of mice after OVA exposure and leonuride treatment. Gene sets belonging to type-2 cytokine/chemokine activity stood out in leonuride target transcripts. Those upregulated (Bmp10, Ccl12, Ccl22, Ccl8, Ccl9, Cxcl15, Il13, Il33, Tnfrsf9, Il31ra, Il5ra, Il13ra2 and Ccl24) or downregulated (Acvr1c and Il18) genes in asthmatic mice, were all reversely regulated by leonuride treatment.. Our results revealed the therapeutic efficacy of leonuride in experimental chronic asthma for the first time, and implied that its anti-inflammatory and antifibrotic properties might be mediated by regulation of type-2 high cytokine/chemokines responses.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Chemokines; Cytokines; Disease Models, Animal; Inflammation; Iridoid Glycosides; Iridoids; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pyrans

Chronic lung diseases are characterized by airway inflammation and remodelling of the lung parenchyma that triggers considerable impairment of respiratory function.. In this study, two compounds belonging to the N-acylhydrazone class were evaluated, aiming to identify new therapeutic agents for pulmonary inflammatory diseases.. The acute toxicity of 2-cyano-N'-(3-ethoxy-4-hydroxybenzylidene)- acetohydrazide (JR-12) and N'-benzylidene-2-cyano-3-phenylacrylohydrazide (JR09-Bz) was evaluated. Afterwards, they were tested in models of ovalbumin (OVA)-induced allergic asthma and pleurisy, bleomycin-induced pulmonary fibrosis, in addition to mucolytic activity.. The compounds did not show toxicity at the dose of 2,000 mg/kg, and no animal died. On OVA-induced pleurisy, animals treated with JR-12 or JR09-Bz at a dose of 10 mg/kg (orally) showed significant inhibition of the leukocyte infiltrate in the bronchoalveolar lavage by 62.5% and 61.5%, respectively, compared to the control group. The compounds JR-12 and JR09-Bz were also active in blocking the allergic asthmatic response triggered by OVA, reducing the leukocyte infiltrate by 73.1% and 69.8%, respectively. Histopathological changes and mast cell migration in treated animals with JR-12 or JR09-Bz were similar to treatment with the reference drugs dexamethasone and montelukast. JR-12 and JR09-Bz also reversed airway remodeling in animals on the bleomycin-induced fibrosis model compared to the control group. Furthermore, it was observed that N-arylhydrazone derivatives showed expectorant and mucolytic activities, increasing mucus secretion by 45.6% and 63.8% for JR-12 and JR09-Bz, respectively.. Together, the results show that JR-12 and JR09-Bz showed promising activity against airway inflammation, as well as low toxicity.

Topics: Allergens; Animals; Asthma; Bleomycin; Bronchoalveolar Lavage Fluid; Cytokines; Dexamethasone; Disease Models, Animal; Expectorants; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pleurisy; Pneumonia

Extracellular acidification is a major component of tissue inflammation, including airway inflammation. The extracellular proton-sensing mechanisms are inherent in various cells including airway structural cells, although their physiological and pathophysiological roles in bronchial smooth muscles (BSMs) are not fully understood. In the present study, to explore the functional role of extracellular acidification on the BSM contraction, the isolated mouse BSMs were exposed to acidic pH under contractile stimulation.. The RT-PCR analyses revealed that the proton-sensing G protein-coupled receptors were expressed both in mouse BSMs and cultured human BSM cells. In the mouse BSMs, change in the extracellular pH from 8.0 to 6.8 caused an augmentation of contraction induced by acetylcholine. Interestingly, the acidic pH-induced BSM hyper-contraction was further augmented in the mice that were sensitized and repeatedly challenged with ovalbumin antigen. In this animal model of asthma, upregulations of G protein-coupled receptor 68 (GPR68) and GPR65, that were believed to be coupled with Gq and Gs proteins respectively, were observed, indicating that the acidic pH could cause hyper-contraction probably via an activation of GPR68. However, psychosine, a putative antagonist for GPR68, failed to block the acidic pH-induced responses.. These findings suggest that extracellular acidification contributes to the airway hyperresponsiveness, a characteristic feature of bronchial asthma. Further studies are required to identify the receptor(s) responsible for sensing extracellular protons in BSM cells.

Topics: Acetylcholine; Animals; Asthma; Bronchi; Bronchial Hyperreactivity; Humans; Hydrogen-Ion Concentration; Inflammation; Mice; Mice, Inbred BALB C; Muscle, Smooth; Ovalbumin; Protons; Psychosine; Receptors, G-Protein-Coupled

This study aimed to explore the effects of forkhead box P2 gene (Foxp2) on T-helper 9 (Th9) differentiation in asthmatic mice. An in vivo asthmatic mouse model was induced with ovalbumin (OVA). An in vitro model was established by culturing CD4

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Differentiation; Disease Models, Animal; Forkhead Transcription Factors; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Repressor Proteins; T-Lymphocytes, Helper-Inducer

Asthma is a chronic inflammatory airway disease associated with the airway narrowing and obstruction. Sinapic acid (SA), a hydroxycinnamic acid, possesses various pharmacological properties including antioxidant and anti-inflammatory activity. This research evaluated effects of different doses of SA on murine model of ovalbumin (OVA)-induced allergic asthma.. Allergic asthma induced by sensitizing mice on days 1 and 14 by intraperitoneal injection of OVA. After initial sensitization, between days 21 and 23, mice were challenged for 30 min with an aerosol of 1 % (wt/vol) OVA. Treatment with dexamethasone (3 mg/kg) or SA (25, 50 or 100 mg/kg) were done by oral gavage on days 15-23. Inflammatory cells infiltration and interferon-γ (IFN-γ), interlukin-4 (IL-4), IL-5 and IL-13 levels were evaluated in the bronchoalveolar lavage fluid (BALF). Serum total and OVA-specific immunoglobulin E (IgE) and lung tissue nitric oxide (NO) levels were measured. Histological changes in lung tissue were examined by staining with hematoxylin and eosin (H&E) for cell infiltration, periodic acid-Schiff (PAS) for mucus production and Masson's trichrome for collagen deposition.. Treatment with SA significantly inhibited inflammatory cell infiltration, enhanced IFN-γ level and decreased IL-4, IL-5 and IL-13 levels in BALF. Serum total and OVA-specific IgE levels and NO level in lung tissue were significantly reduced by SA. Histological examination demonstrated that SA significantly attenuated inflammatory cell infiltration and mucus-producing cells in the lung.. These data suggest that SA may be a new therapeutic potential to treat allergic asthma through suppressing T-helper 2 immune responses.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Asthma; Coumaric Acids; Cytokines; Dexamethasone; Disease Models, Animal; Eosine Yellowish-(YS); Hematoxylin; Immunoglobulin E; Inflammation; Interferon-gamma; Interleukin-13; Interleukin-4; Interleukin-5; Mice; Mice, Inbred BALB C; Nitric Oxide; Ovalbumin; Periodic Acid; Th2 Cells

Childhood asthma is a common chronic childhood disease. Branched-chain amino acid transaminase 1 (BCAT1) was reported to be upregulated in chronic airway diseases, while its role in childhood asthma is unclear. Asthma mouse models were established in neonatal mice by 10 µg ovalbumin (OVA) intraperitoneal injection and 3% OVA inhalational challenge. In OVA-challenged mice, BCAT1 levels were upregulated. BCAT1 inhibitor alleviated airway structure and inflammation by suppressing IgE, OVA-specific IgE and inflammatory cytokine release and inflammatory cell infiltration. BCAT1 inhibitor alleviated airway remodeling by inhibiting goblet cell hyperplasia, mucus secretion and the expression of α-SMA and collagen I/III. The BCAT1 inhibitor prevented OVA-enhanced autophagy by decreasing Beclin-1, Atg5 and LC3I/II and increasing p65 levels. In IL-13-stimulated BEAS-2B cells, rapamycin promoted inflammatory cytokine release and autophagy after BCAT1 inhibitor administration. Our research revealed that BCAT1 was upregulated in neonatal asthmatic mice and that a BCAT1 inhibitor might restrain airway inflammation and remodeling by decreasing autophagy, which offered a novel mechanistic understanding of childhood asthma.

Topics: Airway Remodeling; Amino Acids, Branched-Chain; Animals; Asthma; Autophagy; Beclin-1; Bronchoalveolar Lavage Fluid; Collagen; Cytokines; Disease Models, Animal; Immunoglobulin E; Inflammation; Interleukin-13; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Sirolimus; Transaminases

Phosphatidylinositol 3-kinase gamma (PI3Kγ) has been proven to be a potential target for the treatment of inflammatory diseases of the airway; however, there are few reports of selective PI3Kγ inhibitors being used in the field of airway inflammation thus far. Herein, a study employing in vitro and in vivo methodologies was carried out to assess the anti-airway inflammatory effects of JN-PK1, a selective PI3Kγ inhibitor. In RAW264.7 macrophages, JN-PK1 inhibited PI3Kγ-dependent, cellular C5a-induced AKT Ser473 phosphorylation in a concentration- and time-dependent manner and had no significant effect on cell viability.Furthermore, JN-PK1 significantly suppressed LPS-induced, proinflammatory cytokine expression and nitric oxide production through inhibition of the PI3K signaling pathway in RAW264.7 cells. Then, a murine asthma model was established to evaluate the anti-airway inflammation effect of JN-PK1. BALB/c mice were sensitized and challenged with ovalbumin (OVA) to develop an inflammatory response, fibrosis formation, and other airway changes similar to the symptomatology of asthma in humans. Oral administration of JN-PK1 remarkably attenuated OVA-induced asthma in association with the inhibition of the PI3K signaling pathway. That is to say, the oral administration significantly inhibited increases in inflammatory cell counts and reduced T-helper type 2 cytokine production in bronchoalveolar lavage fluid. Pulmonary histological studies showed that oral administration of JN-PK1 not only reduced the infiltration of inflammatory cells but also retarded airway inflammation and fibration. Taken together, JN-PK1 could be developed as a promising candidate for inflammation therapy, and our findings support some potential for therapeutic inhibition of PI3Kγ to treat inflammatory airway diseases.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors

Gerberae Piloselloidis Herba (GPH), a commonly used traditional medicine in China, is derived from Gerbera piloselloides (Linn.) Cass. It is featured by its special bioactivities as antitussive, expectorant, anti-asthma, anti-bacterial, anti-tumor, uterine analgesia, and immunity-enhancing. With a long history of medication in ethnic minority areas in China, it is often used as an effective treatment for cough and sore throat as well as allergic asthma. Although our previous investigation also has discovered GPH performed effective treatment on allergic asthma, its underlying mechanism remains unclear.. This research aims to reveal the pharmacological mechanism of GPH in the treatment for allergic asthma through combination of plasma pharmacology and network pharmacology.. Firstly, the components of GPH in blood samples were identified using UHPLC- Q-Orbitrap HRMS. An interaction network of "compound-target-disease" was constructed based on the compounds confirmed in blood and on their corresponding targets of allergic asthma acquired from disease gene databases, predicting the possible biological targets and potential signal pathways of GPH with the network pharmacology analysis. Then, a molecular docking between the blood ingredients and the core targets was carried out using the Autodock Vina software. Subsequently, after establishing a mouse model with allergic asthma induced by ovalbumin (OVA), the effect of GPH on allergic asthma was evaluated by analyzing a series of indicators including behavior, lung pathological changes, inflammatory factors in serum and bronchoalveolar lavage fluid (BALF). Finally, the key pathway and targets predicted by network pharmacology and molecular docking were further verified using Western blot analysis.. Eleven chemical constituents (such as arbutin, neochlorogenic acid, chlorogenic acid, etc.) were identified through the analysis of plasma samples, on which basis a total of 142 genes intersecting GPH and allergic asthma were collected by network pharmacology. After performing enrichment analysis of these genes in gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG), it was found that arbutin-related targets mainly focused on phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) signal pathway, while luteolin and marmesin -related targets tended to locate at Interleukin-17 (IL-17) signal pathway. Meanwhile, the findings of molecular docking suggested that such components as arbutin, luteolin and marmesin entering into blood had good binding with the core targets related to PI3K/Akt and IL-17 pathways. In addition, GPH improved the OVA-induced asthma symptoms, the alveolar septa thickening and the infiltration of inflammatory cell around bronchi and bronchioles as well as reduced the levels of IgE, IL-8 and TNF-α in serum or BALF. Furthermore, GPH could inhibit the phosphorylation level of Akt and the expression of PI3K, an efficacy supported by the findings by way of Western blot which suggests that GPH in the treatment of allergic asthma was linked to PI3K/Akt signal pathway.. In this study, a comprehensive strategy to combine the UPLC-Q-Orbitrap HRMS with network pharmacology was employed to clarify the mechanism of GPH against allergic asthma, a finding where GPH may inhibit PI3K/Akt signal pathway to protect mice from OVA-induced allergic asthma. This study provides a deeper understanding of the pharmacological mechanism of GPH in treatment of asthma, offering a scientific reference for further research and clinical application of GPH in terms of allergic asthma.

Topics: Animals; Arbutin; Asthma; Drugs, Chinese Herbal; Ethnicity; Humans; Interleukin-17; Luteolin; Mice; Minority Groups; Molecular Docking Simulation; Network Pharmacology; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt

Asthma is a high-incidence disease in the world. Oxysophocarpine (OSC), a quinolizidine alkaloid displays various pharmacological functions including anti-inflammation, neuroprotective, anti-virus and antioxidant. Here, we established mice and cell asthmatic model to explore the effects of OSC for asthma treatment. Mice were sensitized and challenged with ovalbumin (OVA) and treated with OSC before challenge. Enzyme-linked immuno sorbent assay (ELISA), hematoxylin and eosin (H&E), periodic acid-schiff (PAS), tolonium chloride staining and immunohistochemical assay were performed. OSC treatment inhibited inflammatory cell infiltration and mucus secretion in the airway, reduced IgE level in mouse serum and decreased IL-4, IL-5 production in bronchoalveolar lavage fluid (BALF). OSC also reduced the spleen index to regulate immune function. Meanwhile, NCI-H292 cells were induced by lipopolysaccharide (LPS) to simulate airway epithelial injury. OSC pretreatment decreased the IL-6 and IL-8 cytokine levels, mucin 5 AC expression, and mucin 5 AC mRNA level in the cell model. Further, OSC suppressed the phosphorylation of c-Jun N-terminal kinase (JNK), and activator protein 1 (AP-1, Fos and Jun). These findings revealed that OSC alleviated bronchial asthma associated with JNK/AP-1 signaling pathway.

Topics: Alkaloids; Animals; Antioxidants; Asthma; Cytokines; Disease Models, Animal; Eosine Yellowish-(YS); Hematoxylin; Immunoglobulin E; Interleukin-4; Interleukin-5; Interleukin-6; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Molecular Structure; Mucins; Mucus; Ovalbumin; Periodic Acid; Quinolizidines; RNA, Messenger; Tolonium Chloride; Transcription Factor AP-1

Asthma is a chronic allergic respiratory disease with limited treatment options. Emerging findings indicate an important interaction between the gut microbiota and the lungs, and that the development of asthma causes changes in the gut environment. Hylocereus undatus flower (HUF) is a traditional Chinese medicine used in the treatment of pulmonary and intestinal diseases. Our previous studies have demonstrated significant anti-asthmatic and anti-inflammatory activity, but the exact mechanism has not been elucidated. In the current study, we validated the potential therapeutic asthma properties of HUF in vivo using an ovalbumin-induced allergic asthma mouse model. We found that HUF treatment significantly reduced the key features of allergic asthma, including an elevated respiratory rate, inflammatory cell accumulation, airway inflammation, and the expression of pro-inflammatory molecules. Histological analysis of mouse lungs showed that HUF attenuated lung inflammatory cell infiltration. Periodic acid-Schiff staining confirmed the reduced mucus secretion in lung mucosa, and Masson's staining confirmed the reduced collagen deposition in the lungs after HUF treatment. Western blot and immunohistochemistry confirmed that HUF increased lung SIRT1 and reduced p38MAPK, NF-κBp65, and caspase-1 proteins. 16 S rDNA sequencing showed that HUF improved the endostasis of the disrupted gut microbiota composition in asthmatic mice. Surprisingly, an inflammatory response was found in the gut of asthmatic mice, along with alterations in inflammation-associated SIRT1 and caspase-1 proteins, and HUF was able to ameliorate these lesions. In conclusion, these findings suggest that HUF may be a new drug candidate for the treatment of allergic asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Caspases; Cytokines; Disease Models, Animal; Flowers; Gastrointestinal Microbiome; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Sirtuin 1

Interleukin 5 (IL-5) plays crucial roles in type 2-high asthma by mediating eosinophil maturation, activation, chemotaxis and survival. Inhibition of IL-5 signaling is considered a strategy for asthma treatment. Here, we identified MARCH2 and MARCH3 as critical negative regulators of IL-5-triggered signaling. MARCH2 and MARCH3 associate with the IL-5 receptor α chain (IL-5Rα) and mediate its K27-linked polyubiquitination at K379 and K383, respectively, and its subsequent lysosomal degradation. Deficiency of MARCH2 or MARCH3 modestly increases the level of IL-5Rα and enhances IL-5-induced signaling, whereas double knockout of MARCH2/3 has a more dramatic effect. March2/3 double knockout markedly increases the proportions of eosinophils in the bone marrow and peripheral blood in mice. Double knockout of March2/3 aggravates ovalbumin (OVA)-induced eosinophilia and causes increased inflammatory cell infiltration, peribronchial mucus secretion and production of Th2 cytokines. Neutralization of Il-5 attenuates OVA-induced airway inflammation and the enhanced effects of March2/3 double deficiency. These findings suggest that MARCH2 and MARCH3 play redundant roles in targeting IL-5Rα for degradation and negatively regulating allergic airway inflammation.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Eosinophilia; Eosinophils; Inflammation; Interleukin-5; Interleukin-5 Receptor alpha Subunit; Intracellular Signaling Peptides and Proteins; Ligases; Membrane Proteins; Mice; Mice, Inbred BALB C; Ovalbumin; Ubiquitin; Ubiquitin-Protein Ligases

To screen potent terpenoid compounds against allergic inflammation in vitro and in vivo, five terpenoid compounds including menthone, farnesol, oridonin, β-escin and lupeol, were first selected to compare their anti-allergic inflammation potential using mouse lung mast cells in vitro. Among five selected terpenoid compounds, just menthone treatment decreased TNF-α/IL-10 secretion ratios in lipopolysaccharide -stimulated mast cells in vitro. As a result, menthone was further chosen to treat ovalbumin (OVA)-sensitized and challenged BALB/c mice by gavage for 5 weeks. There were six groups including dietary control (DC group, 0 mg menthone/kg b.w./day), 8 (ML group), 40 (MM group) as well as 200 mg menthone/kg b.w./day (MH group) by gavage, positive control (PC group, 3 mg dexamethasone/kg b.w. by gavage before OVA challenge) and non-treatment control (NTC group, normal mice without treatment) in the experiment. Changes of inflammatory mediators, cell distribution, Th1/Th2 and pro-/anti-inflammatory cytokines secretion as well as relative gene expression amounts of six receptors related to allergic inflammation in the lungs and airways were measured. The results showed that middle menthone supplementation (40 mg menthone/kg b.w./day) in vivo decreased protein and eotaxin, but increased Th1 cytokine levels in the bronchoalveolar lavage fluid. Menthone supplementation inhibited eosinophilia, mast cell degranulation, chemokine (C-C motif) receptor 3 (CC receptor 3) and chemokine (C-X-C motif) receptor 1 (CXC receptor 1) gene expression amounts in the lungs, but restored the percentage of monocytes/macrophages. Our results suggest that menthone supplementation may alleviate allergic asthma through regulating airway allergic inflammation, protein overproduction, eosinophils infiltration, Th1/Th2 immune balance, CC receptor 3 and CXC receptor 1 gene expression amounts in the lungs but restoring the percentage of monocytes/macrophages in allergic asthmatic mice.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dietary Supplements; Disease Models, Animal; Inflammation; Lung; Menthol; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Th2 Cells

Increased expression of substance P (SP) and neurokinin-1 receptor (NK1R) has been noticed in patients with allergic rhinitis (AR) and allergic asthma (AA). However, little is known of the expression of SP and NK1R in monocytes and B cells of AR and AA. In the present study, the expression levels of SP and NK1R were determined by flow cytometry and mouse AR and AA models. The results showed that both percentages of SP+ monocytes and SP+ B cells, and mean fluorescence intensity (MFI) of SP in monocytes were elevated in the blood of AA and AR combined with AA (ARA) patients. Similarly, the percentages of NK1R+ monocytes were elevated in the blood of AR, AA, and ARA patients. Allergens Artemisia sieversiana wild allergen extract (ASWE), house dust mite extract (HDME), and Platanus pollen allergen extract (PPE) increased the expression density of SP molecules (determined by MFI) in an individual monocyte of AR patients. HDME and PPE appeared to enhance SP and NK1R expression in the B cells of ARA and AR patients. In the mouse AR and AA models, the percentages of NK1R+ monocytes and B cells were elevated in blood following OVA (ovalbumin) sensitization and challenge. Knocking out the FcεRI molecule completely abolished the OVA-induced upregulation of expression of NK1R in monocytes and B cells of AA mice. In conclusion, upregulated expressions of SP and NK1R may contribute to the pathogenesis of airway allergy.

Topics: Allergens; Animals; Asthma; Mice; Monocytes; Ovalbumin; Receptors, Neurokinin-1; Rhinitis, Allergic; Substance P

A variety of smooth muscle-specific genes and proteins, including SMAD3, BMPR-II, and MRTF, are involved in airway remodeling in asthma. As a receptor of bone morphogenetic protein (BMP) signaling, BMPR-II has important roles in airway remodeling in asthma. However, the underlying mechanism of BMPR-II in airway smooth muscle cells (ASMCs) in asthma remains incomplete.. Wistar rats were intraperitoneally injected with ovalbumin antigen suspension and aluminium hydroxide and, stimulated with ovalbumin nebulized inhalation to constructed asthma model. Primary ASMCs were isolated with collagenase I and identified by testing the α-SMA expression. Quantitative polymerase chain reaction (qPCR) and western blot assay were employed to detect the gene expression. CCK8, Transwell and Fluo-4 A assays were introduced to measure the cell viability, migration and intracellular Ca. This study indicates that the BMPR-II-SMAD3/MRTF signaling pathway is involved in the process of ASMCs remodeling, providing novel avenues for the identification of new therapeutic modalities.

Topics: Airway Remodeling; Aluminum Hydroxide; Animals; Asthma; Bone Morphogenetic Proteins; Cell Proliferation; Collagenases; Myocytes, Smooth Muscle; Ovalbumin; Rats; Rats, Wistar; RNA, Messenger

Remarkable progress has recently been achieved to identify the biological function and potential value of novel therapeutic targets for the effective control of allergic asthma. Interferon (IFN)-λ has been suggested to restrict chronic inflammation in the lungs of asthmatic mice and we sought to determine the contribution of IFN-λ as an asthma therapeutic. We show that inhaled IFN-λ can restrict Th2 and Th17 inflammation in the lungs of asthmatic mice, accompanied with alteration of IL-10 secretion. BALB/C mice were used for an asthmatic mouse model with OVA. Recombinant IFN-λs (IFN-λ

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Immunity; Inflammation; Interferons; Interleukin-10; Interleukin-13; Interleukin-17; Interleukin-4; Interleukin-5; Mice; Mice, Inbred BALB C; Ovalbumin; Th17 Cells; Th2 Cells

To investigate the effects of acupuncture on phosphorylated P38 mitogen-activated protein kinase (p-P38MAPK), intercellular adhesion molecule-1 (ICAM-1), interferon γ (IFN-γ), and eosinophilic granulocytes (EOS) in lung tissue of asthmatic rats, and to explore the mechanism of acupuncture in regulating the apoptosis of EOS.. Clean-grade male Wistar rats were randomly divided into normal, model, dexamethasone and acupuncture groups, 8 rats in each group. The asthmatic model was established by intraperitoneal injection of mixture suspension (1 mL) of 10% ovalbumin and 10% Al(OH)_3+ normal saline, followed by inhalation of atomized 1% ovalbumin solution for 30 min, once daily for 2 weeks to trigger occurrence of asthmatic symptoms. The rats in dexamethasone group were intraperitoneally injected with 0.9 mg/kg dexamethasone since day 15 once a day for two consecutive weeks. In the acupuncture group, bilateral "Feishu" (BL13), "Pishu" (BL20), "Shenshu" (BL23), "Dingchuan" (EX-B1), and "Danzhong" (CV17) were selected for acupuncture treatment once every other day since day 15 for two consecutive weeks. Uniform reinforcing and reducing manipulation was carried out, and the needles were retained for 30 min. The pathological changes of the lung tissue were observed by hematoxylin-eosin (HE) staining. The apoptosis of EOS in the lung tissue of rats was detected by terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. The expression of p-P38MAPK in the lung tissue was detected by Western blot. The protein and mRNA levels of ICAM-1 and IFN-γ in the lung tissue were determined by immunohistochemistry and real-time PCR, respectively.. The results of HE staining showed that the pulmonary alveoli and surrounding tissues were intact with no inflammatory cell infiltration in the normal group. The model group showed massive exudation of inflammatory materials and thickened pulmonary interstitium. The dexamethasone group and acupuncture group showed less damage of the alveolar structure and only a small number of inflammatory cells around the airway. Compared with the normal group, the apoptosis rate of EOS in lung tissue was decreased (. Acupuncture may inhibit the P38MAPK signaling pathway, down-regulate ICAM-1 expression, and up-regulate IFN-γ expression to promote the apoptosis of EOS and reduce EOS aggregation, thus alleviating the inflammatory response of airway in asthma.

Topics: Acupuncture Therapy; Animals; Apoptosis; Asthma; Dexamethasone; Granulocytes; Intercellular Adhesion Molecule-1; Lung; Male; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Rats; Rats, Wistar; RNA, Messenger; Signal Transduction

Allergic rhinitis is one of the common chronic inflammatory diseases of the nasal mucosa. In order to investigate the effect of zinc on ovalbumin induced allergic rhinitis in BALB/C male mice based on NF-KB signaling pathway, thirty BALB/C male mice are randomly divided into three groups: control group, ovalbumin induced allergic rhinitis asthma group and zinc intervention group. The experimental results show that Zinc supplementation in allergic asthma mice with allergic rhinitis correct the immune response of TH2 cells by inhibiting THE NF-KB signaling pathway, reduce the infiltration of inflammatory cells into lung nasal tissue, and reduce airway co-hyperreactivity to improve the clinical symptoms of asthma and play an immune protective role.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Male; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Rhinitis, Allergic; Signal Transduction; Zinc

Topics: Animals; Asthma; Cytokines; Immunity, Innate; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Steroids

Mahuang decoction (MHD) is a classic famous traditional Chinese medicine and has various pharmacological effects, including anti-inflammation and anti-asthma. In this study, we aimed to investigate the potential protective effect of MHD against asthma and elucidated the underlying mechanism.. A mouse model of asthma was induced by ovalbumin (OVA) treatment, and then treated with MHD to evaluate its effect on the asthma. Gain- or loss-of-function approaches were performed in SP1 and FGFR3 to study their roles in asthma via measurement of airway inflammation, airway remodeling and airway smooth muscle cell (ASMC) proliferation-related factors.. MHD reduced airway inflammation and remodeling. Additionally, MHD contributed to diminished expression of SP1, which was shown to repress airway inflammation and remodeling. Furthermore, SP1 bound to the FGFR3 promoter, resulting in the FGFR3 transcription promotion and ASMC proliferation. Conversely, FGFR3 knockdown abolished airway inflammation and remodeling, the mechanism of which was related to suppression of the PI3K/AKT signaling pathway. Meanwhile, MHD hindered airway inflammation and remodeling following asthma by suppressing the SP1/FGFR3/PI3K/AKT axis.. Taken together, MHD may retard airway inflammation and remodeling by suppressing the SP1/FGFR3/PI3K/AKT axis, which contributes to an extensive understanding of asthma and may provide novel therapeutic options for this disease.

Topics: Animals; Asthma; Disease Models, Animal; Drugs, Chinese Herbal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt

Asthma is a common chronic lung disease without absolute treatment, and hypersensitivity reactions and type 2 immune responses are responsible for asthma pathophysiology. ADAM10 as a metalloproteinase transmembrane protein is critical for development of Th2 responses, and levamisole as an anthelmintic drug has immunomodulatory effects, which not only regulates ADAM10 activity but also can suppress the bone marrow and neutrophil production. Therefore, in the present study, nanoparticles were used as a levamisole delivery system to reduce bone marrow suppression, and the immunomodulatory and ADAM10 inhibitory effects of levamisole were studied in allergic asthma. Asthmatic mice were treated with PLGA-levamisole nanoparticles. Then, AHR, BALF, and blood cell counts, levels of the IgG1 subclass, total and OVA-specific IgE, IL2, IL-4, IL-5, IL-10, IL-13, IL-17, IL-25, IL-33, INF-γ, and TNF-α, gene expression of FoxP3, T-bet, RORγt, PU.1, GATA3, FcεRII, CysLT1R, eotaxin, and ADAM10, and lung histopathology were evaluated. PLGA-LMHCl with considered characteristics could control airway hyper-responsiveness, eosinophils in the BALF, levels of immunoglobulins, Th2-, Th9-, and Th17-derived cytokines and pivotal genes, eosinophilic inflammation, hyperplasia of the goblet cell, and hyperproduction of mucus and could increase Th1- and Treg-derived cytokines and also pivotal genes. It could also modulate the ADAM10 activity and had no effect on the number of neutrophils in the bloodstream. The novel safe nanodrug had no side effect on the bone marrow to produce neutrophils and could control the allegro-immuno-inflammatory response of asthma.

Topics: ADAM10 Protein; Amyloid Precursor Protein Secretases; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Forkhead Transcription Factors; Immunoglobulin E; Immunoglobulin G; Immunologic Factors; Interleukin-10; Interleukin-13; Interleukin-17; Interleukin-2; Interleukin-33; Interleukin-4; Interleukin-5; Levamisole; Lung; Membrane Proteins; Mice; Nanoparticles; Nuclear Receptor Subfamily 1, Group F, Member 3; Ovalbumin; Tumor Necrosis Factor-alpha

Treating asthmatic rheumatoid arthritis patients with abatacept has been shown to associate with better control of asthma symptoms. However, the mechanism behind that is not investigated.. Ovalbumin (OVA)- sensitized BALB/c female mice were treated intranasally (IN) or intraperitoneally (IP) with abatacept 4 hrs before the OVA challenge. The effects of abatacept IN or IP on the lungs and blood levels of Tregs and Bregs and their production of immunosuppressive cytokines, were determined using FACS analysis and ELISA assay.. Treating OVA- sensitized asthmatic mice model with abatacept, IN or IP, reduced lung inflammation. IN treatment with abatacept increased the frequency of IL-35 and IL-10 producing Bregs in the lung tissues to a higher level compared to IP treatment. Moreover, the frequency of lungs LAG3+ Tregs was significantly increased following treatment. This was also associated with a reduction in lung tissue and serum IL-17 levels of treated mice.. These results suggest that abatacept by enhancing IL-35+IL-10+ Bregs and LAG3+ Tregs might reverse IL-17 induced lung inflammation during asthma.

Topics: Abatacept; Administration, Intranasal; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Interleukin-10; Interleukin-17; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Benzo(a)pyrene (BaP), an environmental pollutant, is present in high concentrations in urban smog and cigarette smoke and has been reported to promote high mucin 5AC (MUC5AC) expression. Epithelium-derived inflammatory cytokines are considered an important modulator of mucus oversecretion and MUC5AC overexpression. Here, we investigated whether the effect of BaP on MUC5AC overexpression was associated with cytokine autocrine activity in vivo and in vitro.. In vivo, BALB/c mice were treated with ovalbumin (OVA) in the presence or absence of BaP. Allergy-induced mucus production was assessed by Alcian Blue Periodic acid Schiff (AB-PAS) staining. The human airway epithelial cell line NCI-H292 was used in vitro. MUC5AC and transforming growth factor (TGF)-α mRNA levels were assessed with real-time quantitative PCR. The concentration of cytokines was measured by ELISA. The MUC5AC, p-ERK, ERK, p-EGFR and EGFR proteins were detected by Western blotting in cells or by immunohistochemistry in mouse lungs. Small-interfering RNAs were used for gene silencing.. TGF-α was overproduced in the supernatant of NCI-H292 cells treated with BaP. Knockdown of TGF-α expression inhibited the BaP-induced increase in MUC5AC expression and subsequent activation of the EGFR-ERK signalling pathway. Knocking down aryl hydrocarbon receptor (AhR) expression or treatment with an ROS inhibitor (N-acetyl-L-cysteine) could relieve the TGF-α secretion induced by BaP in epithelial cells. In an animal study, coexposure to BaP with OVA increased mucus production, MUC5AC expression and ROS-EGFR-ERK activation in the lung as well as TGF-α levels in bronchoalveolar lavage fluid (BALF). Furthermore, the concentration of TGF-α in BALF was correlated with MUC5AC mRNA levels. Additionally, TGF-α expression was found to be positively correlated with MUC5AC expression in the airway epithelial cells of smokers. Compared with non-smoker asthma patients, TGF-α serum levels were also elevated in smoker asthma patients.. Autocrine TGF-α was associated with BaP-induced MUC5AC expression in vitro and in vivo. BaP induced TGF-α secretion by activating AhR and producing ROS, which led to activation of the EGFR-ERK pathway.

Topics: Animals; Asthma; Benzo(a)pyrene; Cytokines; ErbB Receptors; Humans; Lung; Mice; Mice, Inbred BALB C; Mucin 5AC; Mucus; Ovalbumin; Reactive Oxygen Species; RNA, Messenger; Transforming Growth Factor alpha

Janus kinase 1 (JAK1) is implicated in multiple inflammatory pathways that are critical for the pathogenesis of asthma, including the interleukin (IL)-4, IL-5, IL-13, and thymic stromal lymphopoietin cytokine signaling pathways, which have previously been targeted to treat allergic asthma. Here, we describe the development of AZD0449 and AZD4604, two novel and highly selective JAK1 inhibitors with promising properties for inhalation.. The effects of AZD0449 and AZD4604 in JAK1 signaling pathways were assessed by measuring phosphorylation of signal transducer and activator of transcription (STAT) proteins and chemokine release using immunoassays of whole blood from healthy human volunteers and rats. Pharmacokinetic studies performed on rats evaluated AZD0449 at a lung deposited dose of 52 μg/kg and AZD4604 at 30 µg/kg. The efficacy of AZD0449 and AZD4604 was assessed by evaluating lung inflammation (cell count and cytokine levels) and the late asthmatic response (average enhanced pause [Penh]).. Both compounds inhibited JAK1-dependent cytokine signaling pathways in a dose-dependent manner in human and rat leukocytes. After intratracheal administration in rats, both compounds exhibited low systemic exposures and medium-to-long terminal lung half-lives (AZD0449, 34 hours; AZD4604, 5 hours). Both compounds inhibited STAT3 and STAT5 phosphorylation in lung tissue from ovalbumin (OVA)-challenged rats. AZD0449 and AZD4604 also inhibited eosinophilia in the lung and reduced the late asthmatic response, measured as Penh in the OVA rat model.. AZD0449 and AZD4604 show potential as inhibitors of signaling pathways involved in asthmatic immune responses, with target engagement demonstrated locally in the lung. These findings support the clinical development of AZD0449 and AZD4604 for the treatment of patients with asthma.

Topics: Animals; Asthma; Cytokines; Humans; Janus Kinase 1; Janus Kinase Inhibitors; Lung; Ovalbumin; Rats; Signal Transduction

Lysophosphatidic acid (LPA), an intercellular lipid mediator, is increased in the bronchoalveolar fluids of patients with asthma after allergen exposure. LPA administration exaggerates allergic responses, and the type 2 LPA receptor (LPA

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Lung; Lysophospholipids; Mice; Mice, Inbred BALB C; Mucins; Ovalbumin

Bronchial asthma is a chronic lung disorder, that affects an estimated 262 million people worldwide, thereby, causing a large socio-economic burden. Drug molecules from natural sources have exhibited a good promise in providing an alternative therapy in many chronic ailments. Solasodine, a glycoalkaloid has received an immense interest due to its large pharmacological and industrial value, however, its usefulness in asthma control has not been investigated till date. In this work, solasodine was tested for its ability to reverse several characteristics of bronchial asthma induced by intraperitoneal injection of ovalbumin (OVA) and aluminium hydroxide in experimental rats. Treating asthmatic animals with solasodine (1 mg/kg b.w. or 10 mg/kg b.w.) or dexamethasone (2.5 mg/kg b.w.) reversed OVA-induced airway hyperresponsiveness, infiltration of inflammatory cells and histamine levels in the airways. Furthermore, as compared to OVA-control rats, allergen-induced elevated levels of IgE, nitrites, nitric oxide, and pro-inflammatory mediators, including TNF-α, IL-1β, LTD-4, and Th2-cytokines, particularly, IL-4, IL-5 were remarkably reduced in both bronchoalveolar lavage fluid and blood. These findings are supported by significant protection offered by various treatments against OVA-induced airway inflammation and mast cell degranulation in mesenteric tissues. Further, In-silico molecular docking studies performed to determine inhibitory potential of solasodine at IL-4 and IL-5, demonstrated strong affinity of phytocompound for these receptors than observed with antagonists previously reported. Results of current study imply that solasodine has therapeutic promise in allergic asthma, presumably due to its ability to prevent mast cell degranulation and consequent generation of histamine and Th2-associated cytokines in airways.

Topics: Allergens; Aluminum Hydroxide; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dexamethasone; Disease Models, Animal; Histamine; Humans; Immunity; Immunoglobulin E; Interleukin-4; Interleukin-5; Lung; Mice; Mice, Inbred BALB C; Molecular Docking Simulation; Nitric Oxide; Nitrites; Ovalbumin; Rats; Solanaceous Alkaloids; Tumor Necrosis Factor-alpha

Asthma is a heterogeneous and complex chronic airway disease with a high incidence rate, characterized by chronic airway inflammation. Although the anti-inflammatory effect of zeaxanthin has been demonstrated in various disease models, its explicit role in allergic asthma remains elusive.. An allergic asthma model was established by ovalbumin (OVA) stimulation in BALB/c nude mice. The pathological examination, collagen deposition and expression of α-smooth muscle actin (α-SMA) in lung tissues were determined by hematoxylin and eosin (H&E), MASSON and immunofluorescence staining, respectively. Besides, the effect of zeaxanthin on inflammation and oxidative stress was assessed by the enzyme-linked immunosorbent assay (ELISA) and spectrophotometry measure. Moreover, the underlying mechanism was analyzed by detecting the expression of phosphorylated p38 (p-p38), p38, β-catenin, p-c-Jun N-terminal kinase (p-JNK) and JNK with western blot assays.. The distinct infiltration of inflammatory cells was observed in the OVA-induced asthma mice model with significantly increased concentrations of immunoglobulin E (IgE), interleukin-4 (IL-4), IL-5, IL-13 and eotaxin (p˂0.001), which were prominently reversed by zeaxanthin treatment (p˂0.001). In addition, zeaxanthin treatment decreased the OVA-induced collagen deposition and α-SMA expression. A similar inhibitory effect of zeaxanthin on the oxidative stress was also observed in the OVA-induced asthma mice model, as evidenced by the prominent decrease of malondialdehyde (MDA) concentration and the remarkable increase of superoxide dismutase (SOD), glutathione S transferase (GST) and Glutathione (GSH) concentrations (p˂0.001). Moreover, zeaxanthin introduction markedly reduced the relative expressions of p-p38/p38, β-catenin and p-JNK/JNK in the OVA-induced asthma mice model (p˂0.001), indicating that zeaxanthin suppressed the p38 mitogen-activated protein kinase (p38 MAPK)/β-catenin signaling pathway in the OVA-induced asthma mice model.. Zeaxanthin attenuated OVA-induced allergic asthma in mice via modulating the p38 MAPK/β-catenin signaling pathway.

Topics: Animals; Asthma; beta Catenin; Disease Models, Animal; Inflammation; Mice; Mice, Inbred BALB C; Mice, Nude; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Signal Transduction; Zeaxanthins

Wenyang Pingchuan Formula (WPCF) is an empirical formula for the treatment of acute childhood asthma. However, the "time-effect" relationship of this prescription is not clear. This paper explores the relationship between Janus activated kinase signal transducer and activator of transcriptions (JAK/STAT), nuclear factor-κB (NF-κB), and microRNA (miR-19a), and also preliminarily determines the best time-effect relationship of WPCF in reducing the airway inflammation in asthmatic mice.. 80 BALB/c mice were randomly divided into four groups: control (CON) group, model (MDL) group, dexamethasone (DEX) group, and WPCF group. MDL group was established through intraperitoneal injection of 10% ovalbumin (OVA) and Al(OH). Significant down-regulation of IL-4 and IL-13 expression (P<0.05) and up-regulation of IFN-γ expression (P<0.05) in BALF have been observed for WPCF group compared with the MDL group. The significant down-regulation of miR-19a mRNA and STAT6, p-STAT6, p65, p-p65 proteins (P<0.05) and up-regulation of SOCS1 and Tnfaip3 proteins (P<0.05) in BALF was also observed for WPCF group compared to the MDL group. During the experiment, the weight of the mice in DEX group significantly decreased (P<0.05) compared with the other groups.. WPCF could restore Th1/Th2 balance. The longer the intervention time, the more effective the treatment. The down-regulation of miR-19a mRNA by activating JAK/STAT and NF-κB signal pathways may be a possible mechanism by which WPCF alleviates airway inflammation.

Topics: Animals; Asthma; Inflammation; Interferon-gamma; Interleukin-13; Mice; Mice, Inbred BALB C; MicroRNAs; NF-kappa B; Ovalbumin; RNA, Messenger

To address the role of methyl-CpG-binding domain 2 (MBD2) in the pathogenesis of asthma and its potential as a target for the asthmatic therapy.. Studies were conducted in asthmatic patients and macrophage-specific. Asthmatic patients and mice challenged with OVA exhibited upregulated MBD2 expression in macrophages, especially in alternatively activated (M2) macrophages. In particular, macrophage-specific knockout of. The above data indicate that Mbd2 implicates in the pathogenesis of asthma predominantly by regulating the polarization of M2 macrophages, which supports that Mbd2 could be a viable target for treatment of asthma in clinical settings.

Topics: Animals; Asthma; DNA-Binding Proteins; Humans; Inflammation; Liposomes; Macrophages; Mice; Mice, Knockout; Ovalbumin; RNA, Small Interfering

Asthma affects a large number of people worldwide and is characterized by chronic allergic airway inflammation. Anatabine is a natural alkaloid that is structurally similar to nicotine and found in the Solanaceae family of plants, with anti-inflammatory properties. Consequently, this study aimed to evaluate the potential therapeutic effect of anatabine against asthma.. Ovalbumin was used to induce asthma in rats. Two asthmatic groups were treated with low and high doses of anatabine.. Asthmatic animals experienced increased total leukocyte count and inflammatory cytokines in bronchoalveolar lavage fluid (BALF), bronchitis, and bronchopneumonia associated with mast cell infiltration. Additionally, inducible nitric oxide synthase immunostaining was observed, with decreased pulmonary antioxidant capacity and enzymes and decreased Nrf2 and HO-1 gene expression while increased NFκB-P65 expression. Interestingly, asthmatic animals treated with anatabine at both doses showed dose-dependently decreased inflammatory cells and cytokine levels within BALF reduced inflammation in the airways through decreased mast cell infiltration within lung tissues and increased antioxidant enzymes and Nrf2 and Ho-1 expression levels.. Our results highlight the potential beneficial effect of anatabine against asthma through anti-inflammatory and antioxidant mechanisms. Therefore, anatabine is a promising candidate for pulmonary asthma treatment.

Topics: Alkaloids; Animals; Anti-Inflammatory Agents; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Heme Oxygenase (Decyclizing); Inflammation; Lung; NF-E2-Related Factor 2; Nicotine; Nitric Oxide Synthase Type II; Ovalbumin; Oxidative Stress; Pyridines; Rats; Up-Regulation

Herbal drugs offer an alternative approach for the treatment of diseases like asthma due to low cost and comparatively less adverse effects in contrast to synthetic drugs. Leaves of Quercus leucotrichophora are traditionally used for the treatment of asthma. The study was aimed to assess the anti-asthmatic activity of Quercus leucotrichophora (QL) methanolic (QLME) and aqueous extracts (QLAE) in ovalbumin-(OVA) induced asthma and chemical characterization of QL extract by High Performance Liquid Chromatography-Diode array detector (HPLC-DAD). Animals were inoculated with OVA (i.p) on day 1 and 14 followed by intranasal challenge on 27th and 29th day. Both extracts of QL at 600, 300 and 150 mg/kg and dexamethasone (2 mg/kg) l were administered consecutively from days 15-26 via oral gavage. The QL extracts notably reduced (p < 0.0001-p < 0.05) total and differential leukocyte counts in blood and BALF and serum IgE levels in contrast to disease control. Both extracts and Dex substantially improved activities of superoxide dismutase, catalase, and GSH, while reduced malondialdehyde level in treated mice. Treatment with extracts and Dex caused significant (p < 0.0001-p < 0.05) downregulation of tumor necrosis factor-α, interleukin-4, - 5, - 13, - 6, - 1β, and NF-κB whereas, increased expression of Aquaporin (AQP) 1 and AQP5 in contrast to disease control. It was inferenced from findings that both extract of QL exhibited notable antiasthmatic potential might be due to presence of Daidzein-glucuronic acid, 3-Hydroxyphloretin 6'-hexoside, Catechin, Quercetin, and Kaemferol.

Topics: Animals; Anti-Asthmatic Agents; Aquaporins; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Catalase; Catechin; Chromatography, High Pressure Liquid; Dexamethasone; Disease Models, Animal; Glucuronic Acid; Immunoglobulin E; Interleukin-4; Lung; Malondialdehyde; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Oxidative Stress; Quercetin; Quercus; Superoxide Dismutase; Synthetic Drugs; Tumor Necrosis Factor-alpha

Ecklonia cava is an edible marine brown alga harvested from the ocean that is widely consumed in Asian countries as a health-promoting medicinal food The objective of the present study is to evaluate the anti-asthma mechanism of a new functional food produced by bioprocessing edible algae Ecklonia cava and shiitake Lentinula edodes mushroom mycelia and isolated fractions.. We used as series of methods, including high performance liquid chromatography, gas chromatography, cell assays, and an in vivo mouse assay to evaluate the asthma-inhibitory effect of Ecklonia cava bioprocessed (fermented) with Lentinula edodes shiitake mushroom mycelium and its isolated fractions in mast cells and in orally fed mice.. The in vitro cell and in vivo mouse assays demonstrate the potential value of the new bioprocessed formulation as an anti-inflammatory and anti-allergic combination of natural compounds against allergic asthma and might also ameliorate allergic manifestations of foods, drugs, and viral infections.

Topics: Agaricales; Aluminum Oxide; Animals; Anti-Allergic Agents; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Cytokines; Immunoglobulin E; Inflammation; Interleukin-10; Leukotriene C4; Mice; Mice, Inbred BALB C; Mycelium; Ovalbumin; Phaeophyceae; Prostaglandin D2; Shiitake Mushrooms; Vascular Cell Adhesion Molecule-1

Bisphenol S (BPS) is increasingly being used as an alternative for bisphenol A; however, its health effects remain unclear. We investigated the effects of oral exposure to low-dose BPS on allergic asthma. C3H/HeJ male mice were intratracheally administered with allergen (ovalbumin (OVA), 1 μg/animal) every 2 weeks from 6 to 11 weeks old. BPS was ingested by drinking water at doses equivalent to 0.04, 0.4, and 4 μg/kg/day. We then examined pulmonary inflammation, airway hyperresponsiveness, serum OVA-specific immunoglobulin (Ig) levels, Th2 cytokine/chemokine production, and mediastinal lymph node (MLN) cell activities. Compared with OVA alone, moderate-dose BPS (BPS-M) with OVA significantly enhanced pulmonary inflammation, airway hyperresponsiveness, and OVA-specific IgE and IgG1. Furthermore, interleukin (IL)-5, IL-13, IL-33, and CCL11/Eotaxin protein levels in the lungs increased. Conversely, these allergic responses were reduced in the high-dose BPS+OVA group. In MLN cells, BPS-M with OVA increased the total cell count and activated antigen-presenting cells including conventional dendritic cell subset (cDC2). After OVA restimulation, cell proliferation and Th2 cytokine production (IL-4, IL-5, and IL-13) in the culture supernatant also increased. Therefore, oral exposure to low-dose BPS may exacerbate allergic asthmatic responses by enhancing Th2-polarized responses and activating the MLN cells.

Topics: Allergens; Animals; Asthma; Cytokines; Disease Models, Animal; Drinking Water; Immunoglobulin E; Immunoglobulin G; Interleukin-13; Interleukin-33; Interleukin-4; Interleukin-5; Lung; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Ovalbumin; Phenols; Pneumonia; Respiratory Hypersensitivity; Sulfones; Th2 Cells

Background and Objectives: Diesel exhaust particulate matter (DEPM) is an air pollutant that is associated with asthma. In this study, the therapeutic efficacy of Weissella cibaria strains CMU (Chonnam Medical University) and CMS (Chonnam Medical School) 1, together with the drug Synatura, an anti-tussive expectorant, was investigated in a murine asthma model exacerbated by DEPM. Materials and Methods: BALB/c mice were sensitized with ovalbumin (OVA) before intranasal challenge with OVA and DEPM. W. cibaria CMU, CMS1, and Synatura were administered orally for 21 days. Results: Neither Synatura nor W. cibaria strains affected spleen, liver, or lung weights. W. cibaria strains CMU and CMS1 significantly reduced the levels of interleukin (IL)-4, OVA-specific immunoglobulin E (IgE), and total lung collagen in bronchoalveolar lavage fluid (BALF), similar to those with Synatura, regardless of the oral dose concentration (p < 0.05). In addition, the W. cibaria CMU strain significantly alleviated IL-1β, IL-6, IL-12, monocyte chemotactic protein-1, and tumor necrosis factor-α in BALF, whereas the CMS1 strain significantly alleviated IL-10 and IL-12 in BALF (p < 0.05); however, Synatura did not show any statistical efficacy against them (p > 0.05). All concentrations of W. cibaria CMU and low concentrations of W. cibaria CMS1 significantly reduced lung bronchiolar changes and inflammatory cell infiltration. Conclusions: In conclusion, W. cibaria CMU in asthmatic mice showed better efficacy than W. cibaria CMS1 in improving asthma exacerbated by DEPM exposure, as well as better results than pharmaceuticals.

Topics: Air Pollutants; Animals; Asthma; Chemokine CCL2; Cytokines; Disease Models, Animal; Expectorants; Humans; Immunoglobulin E; Inflammation; Interleukin-10; Interleukin-12; Interleukin-6; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Tumor Necrosis Factor-alpha; Vehicle Emissions; Weissella

Bronchial asthma is a chronic inflammatory airway illness. For the first time, we evaluated the proposed anti-asthmatic protective and therapeutic potency of inhaling Punica granatum juice (PJE) and peel (PPE) extract mixture (PM). Rats were challenged with ovalbumin (OVA) for 23 days and aerosolized with PM before each OVA challenge (protected group) or following the final OVA challenge for 3 days (therapeutic group). Considerable concentrations of phenolics were detected in PJE and PPE. Therefore, PM demonstrated synergistic scavenging abilities of NO and DPPH radicals. It also showed synergistic anti-inflammatory activities against lipopolysaccharide (LPS)-induced inflammation in the white blood cells by lowering the gene expression of CXCR1, CXCR2, IL-6, and IL-8. In addition, PM increased IL-10 gene expression while decreasing NO and TNF-α levels in LPS-exposed cells. Regarding the rats that were protected with PM, they exerted pulmonary pro-oxidant effects but prevented the OVA-induced upregulation of NF-κB, IKK, TNF-α, COX-2, iNOS, IL-13, and COL1A1, as well as MUC5AC and mucin over-secretion. While PM in the therapeutic group improved reactive oxygen species levels and normalized most of the investigated inflammatory and fibrotic mediators and mucin formation, but slightly improved the antioxidant indices. In addition, OVA-induced morphological alterations were massively improved after PM inhalation for short or long periods. Thus, PM inhalation prevented and treated OVA-induced pulmonary inflammation and fibrosis, while the inhalation period between 3 and 23 days needs to be optimized to acquire a better impact on the antioxidant indices.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Cyclooxygenase 2; Disease Models, Animal; Interleukin-10; Interleukin-13; Interleukin-6; Interleukin-8; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Mucins; NF-kappa B; Ovalbumin; Pomegranate; Rats; Reactive Oxygen Species; Respiratory Aerosols and Droplets; Signal Transduction; Tumor Necrosis Factor-alpha

This study aims to investigate mechanism of "Ephedrae Herba-Descurainiae Semen Lepidii Semen" combination(MT) in the treatment of bronchial asthma based on network pharmacology and in vivo experiment, which is expected to lay a theoretical basis for clinical application of the combination. First, the potential targets of MT in the treatment of bronchial asthma were predicted based on network pharmacology, and the "Chinese medicine-active component-target-pathway-disease" network was constructed, followed by Gene Oncology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment of the potential targets. Molecular docking was used to determine the binding activity of key candidate active components to hub genes. Ovalbumin(OVA, intraperitoneal injection for sensitization and nebulization for excitation) was used to induce bronchial asthma in rats. Rats were classified into control group(CON), model group(M), dexamethasone group(DEX, 0.075 mg·kg~(-1)), and MT(1∶1.5) group. Hematoxylin and eosin(HE), Masson, and periodic acid-Schiff(PAS) staining were performed to observe the effect of MT on pathological changes of lungs and trachea and goblet cell proliferation in asthma rats. The levels of transforming growth factor(TGF)-β1, interleukin(IL)6, and IL10 in rat serum were detected by enzyme-linked immunosorbent assay(ELISA), and the mRNA and protein levels of mitogen-activated protein kinase 8(MAPK8), cyclin D1(CCND1), IL6, epidermal growth factor receptor(EGFR), phosphatidylinositol 3-kinase(PI3 K), and protein kinase B(Akt) by qRT-PCR and Western blot. Network pharmacology predicted that MAPK8, CCND1, IL6, and EGFR were the potential targets of MT in the treatment of asthma, which may be related to PI3 K/Akt signaling pathway. Quercetin and β-sitosterol in MT acted on a lot of targets related to asthma, and molecular docking results showed that quercetin and β-sitosterol had strong binding activity to MAPK, PI3 K, and Akt. In vivo experiment showed that MT could effectively alleviate the symptoms of OVA-induced asthma rats, improve the pathological changes of lung tissue, reduce the production of goblet cells, inhibit the inflammatory response of asthma rats, suppress the expression of MAPK8, CCND1, IL6, and EGFR, and regulate the PI3 K/Akt signaling pathway. Therefore, MT may relieve the symptoms and inhibit inflammation of asthma rats by regulating the PI3 K/Akt signaling pathway, and quercetin and β-sitos

Topics: Animals; Asthma; Cyclin D1; Dexamethasone; Drug Combinations; Drugs, Chinese Herbal; Eosine Yellowish-(YS); Ephedra; ErbB Receptors; Hematoxylin; Interleukin-10; Interleukin-6; Mitogen-Activated Protein Kinase 8; Molecular Docking Simulation; Network Pharmacology; Ovalbumin; Periodic Acid; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Quercetin; Rats; RNA, Messenger

The role of the calcium-sensitive receptor (CaSR) was assessed in a juvenile mouse model of asthma induced by ovalbumin (OVA). The experiment was divided into normal control, OVA, and OVA +2.5/5 mg/kg NPS2143 (a CaSR antagonist) groups. OVA induction was performed in all groups except the normal control, followed by assessing airway hyperresponsiveness (AHR) and lung pathological changes. Serum OVA-specific IgE and IgG1 were detected with an enzyme-linked immunosorbent assay (ELISA), and inflammatory cells were counted in bronchoalveolar lavage fluid (BALF). Real-time quantitative polymerase chain reaction, ELISA, and western blotting were performed to detect gene and protein expression. NPS2143 improved the OVA-induced AHR in mice, and AHR was higher in the OVA +2.5 mg/kg NPS2143 group than in the OVA +5 mg/kg NPS2143 group. Furthermore, NPS2143 reduced the production of OVA-specific IgE and IgG1 in serum and the number of eosinophils and lymphocytes in BALF in OVA mice with reduced CaSR expression in lung tissues. Besides, OVA-induced mice exhibited peribronchial and perivascular inflammatory cell infiltration, which was accompanied by severe goblet cell hyperplasia/hyperplasia and airway mucus hypersecretion. Furthermore, these mice exhibited increased levels of Interleukin (IL)-5, IL-13, MCP-1, and eotaxin, which were alleviated by NPS2143. The 5 mg/kg NPS2143 showed more effective than the 2.5 mg/kg treatment. CaSR expression was elevated in the lung tissues of OVA-induced asthmatic juvenile mice, whereas the CaSR antagonist NPS2143 reduced AHR and attenuated the inflammatory response in OVA-induced juvenile mice, possibly exerting therapeutic effects on childhood asthma.

Topics: Animals; Asthma; Calcium; Disease Models, Animal; Hyperplasia; Immunoglobulin E; Immunoglobulin G; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Background Dysregulation of the balance between different T cell populations is believed to be an important basis for asthma.Objective To observe the changes in γδT subtypes in transgenic asthmatic mice after aerosol inhalation of Mycobacterium vaccae, and to further investigate the mechanism of M. vaccae in asthmatic mice and its relationship with γδT cells.Methods TCR-β-/- mice were exposed to atomized normal saline or M. vaccae for 5 days and the γδT cells from the lung tissues were isolated. Changes in γδT17 and γδTreg populations were detected. Asthma was induced in BALB/c mice using ovalbumin, which was then transplanted with control or M. vaccae-primed γδT cells. First we analyzed the content of γδT cells that secrete IL-17 (IL-17 γδT cells) and Foxp3+ γδT cells in lung tissues and then measured the content of IL-17 in the bronchoalveolar lavage fluid (BALF) by ELISA.Results Exposure to M. vaccae increased and decreased the relative proportions of Foxp3+ γδT cells and IL-17+ γδT cells, respectively, thereby decreasing airway reactivity and inflammation levels in asthmatic mice, and significantly decreasing IL-17 levels in BALF. Furthermore, mice treated with these primed T cells showed a decrease in IL-17+ γδT cells, and a concomitant increase in Foxp3+ γδT cells in their lung tissues. Furthermore, adoptive transfer of M. vaccae-primed γδT cells decreased GATA3 and NICD and increased T-bet in lung.Conclusions The M. vaccae-primed γδT cells alleviated the symptoms of asthma by reversing Th2 polarization in the lungs and inhibiting the Notch/GATA3 pathway.

Topics: Animals; Asthma; Disease Models, Animal; Forkhead Transcription Factors; Interleukin-17; Lung; Mice; Mice, Inbred BALB C; Mycobacteriaceae; Ovalbumin; Receptors, Antigen, T-Cell; Saline Solution; T-Lymphocytes, Regulatory; Th17 Cells

Asthma is a disorder characterized by airflow obstruction, inflammation, declining airway function, bronchial hyperresponsiveness and tissue remodelling. Probiotics are defined as "live microorganisms that, when administered in adequate amounts, confer a health benefit on the host". The use of probiotics is becoming increasingly studied and recent evidence has suggested that it may provide therapeutic benefits in asthma and other diseases. Lactobacillus delbrueckii UFV-H2b20 fulfils all the requirements to be classified as probiotic. Previous studies have already shown the ability of L. delbrueckii UFV-H2b20 to stimulate the immune system. Our objective was to evaluate the protective effects of L. delbrueckii UFV-H2b20 in experimental allergic asthma. We used a murine model of ovalbumin-induced allergic airway inflammation to mimic allergic asthma. Oral treatment with L. delbrueckii UFV-H2b20 improves respiratory parameters and inhibits the inflammatory response in the lungs by decreasing the numbers of inflammatory monocytes, eosinophils and alveolar macrophages, as well as IgE levels. Treatment increased the IFN-γ/IL-4 cytokine ratio. Levels of IL-10 in the lungs were also increased in treated animals. Our results also showed that the probiotic administration increases the number of CD39

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Disease Models, Animal; Inflammation; Interferon-gamma; Lactobacillus delbrueckii; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; T-Lymphocytes, Regulatory

Infants with respiratory syncytial virus (RSV)-associated bronchiolitis are at increased risk of childhood asthma. Recent studies demonstrated that certain infections induce innate immune memory (also termed trained immunity), especially in macrophages, to respond more strongly to future stimuli with broad specificity, involving in human inflammatory diseases. Metabolic reprogramming increases the capacity of the innate immune cells to respond to a secondary stimulation, is a crucial step for the induction of trained immunity. We hypothesize that specific metabolic reprogramming of lung trained macrophages induced by neonatal respiratory infection is crucial for childhood allergic asthma.. To address the role of metabolic reprogramming in lung trained macrophages induced by respiratory virus infection in allergic asthma.. Neonatal mice were infected and sensitized by the natural rodent pathogen Pneumonia virus of mice (PVM), a mouse equivalent strain of human RSV, combined with ovalbumin (OVA). Lung CD11b. Proline metabolism reprogramming of trained macrophages induced by early respiratory infection combined with allergen sensitization contributes to development of allergic asthma in childhood. Proline metabolism could be a well target for prevention of allergic asthma in childhood.

Topics: Allergens; Animals; Asthma; Humans; Hypersensitivity; Macrophages; Mice; Mice, Inbred BALB C; Ovalbumin; Proline; Respiratory Syncytial Virus Infections; Respiratory Tract Infections

The aim of the study was to determine the role and mechanism of runt-related transcription factor 3 (Runx3) in the development of asthma.. An asthma mouse model was constructed and validated by hematoxylin-eosin analysis of lung tissue and noninvasive enhanced pause (Penh) evaluation of airway hyperresponsiveness. Then, the levels of Runx3 and interleukin (IL)-12 in peripheral blood and lung tissue were detected by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. By use Runx3+/- mice, the effect of Runx3 downregulation on ovalbumin (OVA)-induced asthma was investigated. After stimulated by different doses of IL-12, the expressions of Runx3, hypoxia inducible factor-1α (HIF-1α), and NOD-like receptor thermal protein domain associated protein 3 (NLRP3) in BEAS-2B cells were tested through Western blot and immunofluorescence. Subsequently, BEAS-2B cells treated with 20 ng/mL IL-12 were divided into control, Runx3 overexpression negative control, Runx3 overexpression, HIF-1α inhibitor, and Runx3 overexpression + HIF-1α agonist groups. The Western blot, immunofluorescence, and ELISA indicators were tested repeatedly.. The increased number of inflammatory cells and Penh value confirmed the success of the asthma mouse model. IL-12 expression was significantly increased, and Runx3 was reduced in asthma mice compared with wild-type mice. Meanwhile, the level of immunoglobulin E (IgE) in serum, cytokines in bronchoalveolar lavage fluid, and IL-12, HIF-1α, NLRP3 in the lung were significantly elevated in Runx3+/- mice. With the increase of IL-12 concentration, Runx3 protein expression decreased, while HIF-1α and NLRP3 expression increased. Further mechanistic studies suggest that Runx3 ameliorates IL-12-induced BEAS-2B injury by inhibiting HIF-1α/NLRP3 pathway.. These results suggested that IL-12 contributes to the development of asthma by targeting HIF-1α/NLRP3 pathway through Runx3, thus providing a novel strategy for asthma therapy.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Interleukin-12; Lung; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Signal Transduction

Asthma is one of the most common inflammatory diseases of the lung worldwide. There has been considerable progress in recent studies to treat and prevent allergic asthma, however, various side effects are still observed in clinical practice. Six-week-old male BALB/c mice were orally administered with either sword bean pod extracts (SBP; 100 or 300 mg/kg) or dexamethasone (DEX; 5 mg/kg) once daily over 3 weeks, followed by ovalbumin sensitization (OVA/Alum.; intraperitoneal administration, 50 μg/2 mg/per mouse). Scoring of lung inflammation was performed to observe pathological changes in response to SBP treatment compared to OVA/Alum.-induced lung injury. Additionally, inflammatory cytokines were quantified in serum, bronchoalveolar lavage fluid (BALF), and lung tissue using ELISA and Western blot analyses. SBP treatment significantly reduced the infiltration of inflammatory cells, and release of histamine, immunoglobulin E, and leukotriene in serum and BALF. Moreover, the therapeutic effect of SBP was also assessed to analyze the inflammatory changes in the lung tissues. SBP markedly suppressed the activation of the MAPK signaling pathway and the expression of key inflammatory proteins (e.g., TNF-α) and Th2 type cytokines (IL-5 and IL-13). SBP was effective in ameliorating the allergic inflammation against OVA/Alum.-induced asthma by suppressing pulmonary inflammation.

Topics: Alum Compounds; Animals; Asthma; Bronchoalveolar Lavage Fluid; Canavalia; Cytokines; Dexamethasone; Disease Models, Animal; Histamine; Immunoglobulin E; Inflammation; Interleukin-13; Interleukin-5; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Pneumonia; Tumor Necrosis Factor-alpha

The role of micro RNAs (miRNAs) in asthma remains unclear. In this study, we examined the role of miRNA in targeting FOXO1 in asthma. Results showed that miR-493-5p was one of the differentially expressed miRNAs in the PBMCs of asthmatic children, and was also associated with Th cell differentiation. The miR-493-5p expression decreased significantly in the OVA-induced asthma mice than the control groups. The miR-493-5p mimic inhibited the expression of the IL-9, IRF4 and FOXO1, while the inhibitor restored these effects. Moreover, the Dual-Luciferase analysis results showed FOXO1 as a novel valid target of miR-493-5p. According to the rescue experiment, miR-493-5p inhibited Th9 cell differentiation by targeting FOXO1. Then the exosomes in association with the pathogenesis of asthma was identified. Various inflammatory cells implicated in asthmatic processes including B and T lymphocytes, DCs, mast cells, and epithelial cells can release exosomes. Our results demonstrated that the DC-derived exosomes can inhibit Th9 cell differentiation through miR-493-5p, thus DC-derived exosomal miR-493-5p/FOXO1/Th9 may serve as a potential therapeutic target in the development of asthma.

Topics: Animals; Asthma; Cell Differentiation; Forkhead Box Protein O1; Interleukin-9; Mice; MicroRNAs; Ovalbumin; T-Lymphocytes, Helper-Inducer

Gestational arsenic (As) exposure has been associated with adverse developmental outcomes. The purpose of this study was to explore the impacts of As exposure in different periods on susceptibility to allergic asthma. In model 1, dams were administered with NaAsO

Topics: Animals; Arsenic; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Drinking Water; Female; Goblet Cells; Hyperplasia; Lung; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Pregnancy

Asthma is one of the most common chronic diseases that affects more than 300 million people worldwide. Though most asthma can be well controlled, individuals with severe asthma experience recurrent exacerbations and impose a substantial economic burden on healthcare system. Neutrophil inflammation often occurs in patients with severe asthma who have poor response to glucocorticoids, increasing the difficulty of clinical treatment.. We established several neutrophil-dominated allergic asthma mouse models, and analyzed the airway hyperresponsiveness, airway inflammation and lung pathological changes. Neutrophil extracellular traps (NETs) formation was analyzed using confocal microscopy and western blot.. We found that the ovalbumin (OVA)/complete Freund's adjuvant (CFA)/low-dose lipopolysaccharide (LPS)-induced mouse model best recapitulated the complex alterations in the airways of human severe asthmatic patients. We also observed OVA/CFA/LPS-exposed mice produced large quantities of neutrophil extracellular traps (NETs) in lung tissue and bone marrow neutrophils. Furthermore, we found that reducing the production of NETs or increasing the degradation of NETs can reduce airway inflammation and airway hyperresponsiveness.. Our findings identify a novel mouse model of neutrophilic asthma. We have also identified NETs play a significant role in neutrophilic asthma models and contribute to neutrophilic asthma pathogenesis. NETs may serve as a promising therapeutic target for neutrophilic asthma.

Topics: Animals; Asthma; Disease Models, Animal; Freund's Adjuvant; Glucocorticoids; Humans; Inflammation; Lipopolysaccharides; Mice; Neutrophil Activation; Ovalbumin; Respiratory Hypersensitivity

The traditional Chinese medicine (TCM) formula Shegan Mahuang Decoction (SMD) has been used for treating asthma with significant clinical efficacy, but its mechanism of action has not been well investigated. This study aimed to investigate the anti-asthma effects of SMD on ovalbumin (OVA)-induced airway hyperresponsiveness (AHR) in rats and its potential mechanisms using liquid chromatography-mass spectrometry (LC-MS)-based metabolomics combined with Gene Expression Omnibus (GEO) data mining. The results showed that the administration of SMD significantly attenuated OVA-induced lung histopathological changes. OVA-induced elevation of the immunoglobulin (IgE) and interleukin-4 (IL-4) levels was also inhibited by SMD. A total of 28 significantly changed metabolites in plasma were selected from metabolomics analysis. After treatment with SMD, 24 of them were negatively regulated and the related metabolisms were involved in multiple metabolic pathways such as sphingolipid metabolism and arachidonic acid metabolism. The differentially expressed genes (DEGs) were obtained by GEO data mining. The integrated pathway analysis highlighted 11 signaling pathways that were associated with the anti-asthma effect of SMD. Among them, the metabolite-gene-pathway network showed that the peroxisome proliferator-activated receptors (PPAR) signaling pathway might be the most significant one. This study revealed that SMD exerted an anti-asthma effect against OVA-induced AHR

Topics: Animals; Anti-Asthmatic Agents; Arachidonic Acid; Asthma; Bronchoalveolar Lavage Fluid; Chromatography, Liquid; Cytokines; Drugs, Chinese Herbal; Ovalbumin; Peroxisome Proliferator-Activated Receptors; Rats; Sphingolipids; Tandem Mass Spectrometry

Asthma is an inflammatory syndrome characterized by airway hyperresponsiveness, bronchial inflammation, and airway remodeling. Abnormal proliferation of airway smooth muscle cells (ASMCs) is the main pathological feature of asthma. This study investigated the function and mechanism of serine arginine-rich splicing factor 1 (SRSF1) in ASMC proliferation in asthma.. SRSF1 expressions in the bronchi of ovalbumin-induced asthmatic mice and IgE-treated mouse ASMCs (mASMCs) were evaluated using quantitative real-time PCR and Western blot. The localization and expression of SRSF1 in the bronchi of asthmatic mice were assessed by immunohistochemistry. Functionally, gain- and loss-of-function assays, flow cytometry, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays were conducted. Mechanistically, RNA degradation assay, RNA immunoprecipitation, RNA pull-down, and dual-luciferase reporter gene assays were carried out.. SRSF1 was highly expressed in the bronchi of ovalbumin-induced asthma mice and IgE-treated mASMCs and was mainly located in the nucleus. Experiments on the function of SRSF1 showed that the silencing of SRSF1 induced the cell cycle of mASMC arrest and restrained mASMC proliferation. Investigations into the mechanism of SRSF1 revealed that SRSF1 and miR-135a are competitively bound to the 3'UTR region of Cyclin D2 (CCND2). SRSF1 overexpression repressed the degradation of CCND2 mRNA, and miR-135a negatively regulated CCND2 expression. Furthermore, SRSF1 knockdown inhibited ASMC proliferation in asthma mouse models by regulating the levels of miR-135a and CCND2.. SRSF1 knockdown repressed ASMC proliferation in asthma by regulating miR-135a/CCND2 levels.

Topics: Animals; Asthma; Bronchi; Cell Proliferation; Cyclin D2; Immunoglobulin E; Mice; MicroRNAs; Myocytes, Smooth Muscle; Ovalbumin; Serine-Arginine Splicing Factors

Imperatorin is a furanocoumarin derivative and an effective ingredient in several Chinese medicinal herbs. It has favorable expectorant, analgesic, and anti-inflammatory effects. In this study, we investigated whether imperatorin has protective effects against

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dermatophagoides pteronyssinus; Disease Models, Animal; Eosine Yellowish-(YS); Expectorants; Furocoumarins; Hematoxylin; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-10; Interleukin-12; Interleukin-13; Interleukin-5; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Epidemiological investigations show that long-term exposure to PM2.5 is directly related to asthma-like and other respiratory diseases. This study aims to further explore the pharmacological effect of Ephedra sinica polysaccharide (ESP) on lung injury caused by atmospheric PM2.5.. To achieve the aim, we explored the therapeutic effect of ESP on an aggravated asthma-like mouse induced by PM2.5 combined with ovalbumin (OVA), and explored mechanisms underlying the connection between gut microbiota and lung function.. Preliminary results showed that ESP alleviated the symptoms of aggravated allergic asthma-like in mice; reduced the number of eosinophils in BALF; reduced the levels of serum Ig-E, IL-6, TNF-α, and IL-1β. Further qRT-PCR detected that ESP inhibited the NF-κB pathway. The final analysis detected by 16S rRNA and short chain fatty acid (SCFA) confirmed that ESP increased relative proportions of Bacteroides, Lactobacillus, Prevotella, Butyricicoccus and Paraprevotella, but decreased that of Enterococcus and Ruminococcus; increased acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, and isohexanic acid in the meanwhile.. The study showed that ESP has a potential for future therapeutical applications in the prevention and treatment of asthma-like disease induced by PM2.5 and OVA via regulation of gut microbiota and SCFA.

Topics: Animals; Asthma; Disease Models, Animal; Ephedra sinica; Fatty Acids, Volatile; Gastrointestinal Microbiome; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Polysaccharides; RNA, Ribosomal, 16S

Asthmatics exhibit clinical fluctuations between manageable and treatment-resistant phenotypes as a worldwide socioeconomic health burden. Sonic Hedgehog (Shh) genes mediate regulatory pulmonary cell renewal in adults and contribute to the pathogenesis of high phenotypic asthma which depends mainly on T helper-2 (Th-2) cells and related cytokines. However, the exact pathophysiological roles of Shh molecular signalling in the Th-17-dependent low phenotypic allergic airway inflammation and asthma are not evidenced previously.. Ovalbumin (OVA) and OVA/lipopolysaccharide (LPS)-sensitized and challenged BALB/c mice were enrolled currently to assess the Shh signalling proteins. Furthermore, the effects of vismodegib, a Smo inhibitor, on the modulation of Shh signalling were compared to dexamethasone. The asthma phenotypes were confirmed by serum total immunoglobulin-E (IgE), bronchoalveolar lavage (BAL) fluid white blood cell counts, lung interleukins, tumor necrosis factor (TNF)-α, transforming growth factor (TGF)-β1, and histopathological changes, and scoring.. Mice challenged with OVA or OVA/LPS showed upregulated lung Shh, patched (Ptch1), smoothened (Smo), and Gli1 proteins. Vismodegib in the two experimental phenotypes of asthma showed reduced airway inflammation and remodelling. Additionally, vismodegib reduced the eosinophilia and neutrophilia reported in high and low asthma types, respectively. Moreover, vismodegib and dexamethasone exhibited negative feedback control throughout the enhanced Shh signalling cascades, including Shh, Ptch1, and Gli1 in several asthma models.. In conclusion, Shh signalling partially elucidates the OVA/LPS-challenged mice with severe asthma, which proposes a new promising molecular therapeutic target. Furthermore, Smo inhibition by vismodegib has therapeutic potential in both experimental eosinophilic and neutrophilic allergic airway diseases.

Topics: Anilides; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dexamethasone; Disease Models, Animal; Hedgehog Proteins; Inflammation; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pyridines; Zinc Finger Protein GLI1

Asthma is a highly prevalent and heterogeneous chronic respiratory disease and is often treated with inhaled corticosteroids or in combination with a β

Topics: Adrenergic beta-Agonists; Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Disease Models, Animal; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Perilla frutescens; Pneumonia; Receptors, Adrenergic, beta; Rosmarinic Acid; Signal Transduction

Cycloastragenol (CAG) has been reported to alleviate airway inflammation in ovalbumin- (OVA-) induced asthmatic mice. However, its specific mechanisms remain unclear.. This study is aimed at investigating the effects of CAG on asthma, comparing its efficacy with dexamethasone (DEX), and elucidating the mechanism of CAG's regulation.. The asthma mouse model was induced by OVA. CAG at the optimal dose of 125 mg/kg was given every day from day 0 for 20-day prevention or from day 14 for a 7-day treatment. We observed the preventive and therapeutic effects of CAG in asthmatic mice by evaluating the airway inflammation, AHR, and mucus secretion. Lung proteins were used for TMT-based quantitative proteomic analysis to enunciate its regulatory mechanisms.. The early administration of 125 mg/kg CAG before asthma happened prevented asthmatic mice from AHR, airway inflammation, and mucus hypersecretion, returning to nearly the original baseline. Alternatively, the administration of CAG during asthma also had the same therapeutic effects as DEX. The proteomic analysis revealed that the therapeutical effects of CAG were associated with 248 differentially expressed proteins and 3 enriched KEGG pathways. We then focused on 3 differentially expressed proteins (ITGAL, Syk, and Vav1) and demonstrated that CAG treatment downregulated ITGAL, Syk, and Vav1 by quantitative real-time PCR, western blot analysis, and immunohistochemical staining.. These findings suggest that CAG exerts preventive and protective effects on asthma by inhibiting ITGAL, Syk, and the downstream target Vav1.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Down-Regulation; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Proteomics

Lymphocytes infiltration is a key mechanism that drives asthma lung inflammation. Our previous results demonstrated a significant increase in the frequency and persistence of central memory T (T

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Endothelial Cells; Humans; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Simvastatin; Th2 Cells

Chronic inflammation in the airway passage leads to the clinical syndrome of pediatric asthma. Allergic reactions caused by bacterial, viral, and fungal infection lead to the immune dis-balance which primes T helper cells (Th2), a specific cluster of differentiation 4 (CD4) T cell differentiation. This favors the Th2-specific response by activating the inter-leukin 4/interleukin 13 (IL-4/IL-13) cytokine signaling and further activates the secretion of immunoglobulin E (IgE). IL-13 develops bronchial asthma by elevating bronchial hyperresponsiveness and enables production of immunoglobulin M (IgM) and IgE. The present study aims to target IL-13 signaling using molecular docking and understanding molecular dynamic simulation (MDS) to propose a compelling candidate to treat asthma. We developed a library of available allergic drugs (n=20) and checked the binding affinity against IL-13 protein (3BPN.pdb) through molecular docking and confirmed the best pose binding energy of -3.84 and -3.71 for epinephrine and guaifenesin, respectively. Studying the interaction of hydrogen bonds and Van der Walls, it is estimated that electrostatic energy is sufficient to interact with the active site of the IL-13 and has shown to inhibit inflammatory signaling. These computational results confirm epinephrine and guaifenesin as potential ligands showing potential inhibitory activity for IL-13 signaling. This study also suggests the designing of a new ligand and screening of a large cohort of drugs, in the future, to predict the exact mechanism to control the critical feature of asthma.

Topics: Animals; Asthma; Child; Cytokines; Disease Models, Animal; Epinephrine; Guaifenesin; Humans; Hypersensitivity; Immunoglobulin E; Interleukin-13; Mice; Mice, Inbred BALB C; Molecular Docking Simulation; Ovalbumin; Th2 Cells

The current asthma therapies are inadequate for many patients with severe asthma. Pyrroloquinoline quinone (PQQ) is a naturally-occurring redox cofactor and nutrient that can exert a multitude of physiological effects, including anti-inflammatory and antioxidative effects. We sought to explore the effects of PQQ on allergic airway inflammation and reveal the underlying mechanisms.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Janus Kinases; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; PQQ Cofactor; Signal Transduction; STAT Transcription Factors; Th2 Cells

Topics: Alkaloids; Amides; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dendritic Cells; Disease Models, Animal; Ephedra sinica; Humans; Lung; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Receptor, PAR-2

Doublecortin-like kinase 1 (DCLK1) has been recognized as a marker of cancer stem cell in several malignancies. Thrombin is crucial in asthma severity as it can promote IL-8/CXCL8 production in lung epithelial cells, which is a potent chemoattractant for neutrophils. However, the pathologic role of DCLK1 in asthma and its involvement in thrombin-stimulated IL-8/CXCL8 expression remain unknown.. IL-8/CXCL8, thrombin, and DCLK1 expression were observed in the lung tissues of severe asthma patients and ovalbumin (OVA)-induced asthmatic mice model. A549 and BEAS-2B cells were either pretreated with inhibitors or small interfering RNAs (siRNAs) before being treated with thrombin. IL-8/CXCL8 expression and the molecules involved in signaling pathway were performed using ELISA, luciferase activity assay, Western blot, or ChIP assay.. IL-8/CXCL8, thrombin, and DCLK1 were overexpressed in the lung tissues of severe asthma patients and ovalbumin (OVA)-induced asthmatic mice model. Our in vitro study found that DCLK siRNA or LRKK2-IN-1 (DCLK1 inhibitor) attenuated IL-8/CXCL8 release after thrombin induction in A549 and BEAS-2B cells. Thrombin activated DCLK1, RhoA, and YAP in a time-dependent manner, in which DCLK1 siRNA inhibited RhoA and YAP activation. YAP was dephosphorylated on the Ser127 site after thrombin stimulation, resulting in YAP translocation to the nucleus from the cytosol. DCLK1, RhoA and YAP activation following thrombin stimulation were inhibited by U0126 (ERK inhibitor). Moreover, DCLK1 and YAP siRNA inhibited κB-luciferase activity. Thrombin stimulated the recruitment of YAP and p65 to the NF-κB site of the IL-8/CXCL8 promoter and was inhibited by DCLK1 siRNA.. Thrombin activates the DCLK1/RhoA signaling pathway, which promotes YAP activation and translocation to the nucleus from the cytosol, resulting in YAP/p65 formation, and binding to the NF-κB site, which enhances IL-8/CXCL8 expression. DCLK1 might be essential in thrombin-stimulated IL-8/CXCL8 expression in asthmatic lungs and indicates a potential therapeutic strategy for severe asthma treatment.

Topics: Animals; Asthma; Doublecortin-Like Kinases; Epithelial Cells; Humans; Interleukin-8; Luciferases; Lung; Mice; NF-kappa B; Ovalbumin; Phosphorylation; Protein Serine-Threonine Kinases; rhoA GTP-Binding Protein; RNA, Small Interfering; Thrombin

Severe neutrophilic asthma is often characterized by persistent airway inflammation and irreversible airway remodeling, which are overstimulated by the high-mobility group box protein 1 (HMGB1). Although wogonin, an O-methylated flavone, has been widely used to treat inflammatory and allergic diseases, its therapeutic effects and potential mechanisms on severe neutrophilic asthma remain elusive.. To evaluate whether wogonin alleviates airway neutrophilia through inducing neutrophil apoptosis and attenuates airway smooth muscle cells (ASMCs) proliferation and migration.. The effect of wogonin on reducing neutrophilic airway inflammation, including neutrophil infiltration and inflammatory mediators, was examined in a mouse model of severe neutrophilic asthma sensitized with ovalbumin and lipopolysaccharide. Also, the effect of wogonin on inducing human neutrophil apoptosis was manifested using cellular morphology, flow cytometry, and caspase inhibition assays. Furthermore, the effect of wogonin on inhibiting HMGB1-mediated ASMCs proliferation and migration was determined.. Wogonin reduced the frequency of neutrophils and inhibited the production of multiple inflammatory mediators, including ovalbumin-specific IgE, tumor necrosis factor-α, interleukin-6, and HMGB1, in bronchoalveolar lavage fluid and lung tissues of the neutrophilic asthmatic mouse model. These data strongly support a significantly suppressed neutrophilic airway inflammation, functionally consistent to the relieved airway hyperresponsiveness by wogonin in vivo. Wogonin induced human neutrophil apoptosis in a dose-dependent manner by activating caspase-8 and caspase-3 in vitro. Wogonin pretreatment abolished HMGB1-induced ASMCs proliferation and migration, which can be explained by the inhibition of phosphorylation in the mitogen-activated protein kinase (MAPK) /Akt singling pathways.. Our findings demonstrate that wogonin augments caspase-dependent apoptosis in neutrophils to alleviate neutrophilic inflammatory responses and regulates intracellular signaling to inhibit HMGB1-mediated ASMCs activation, providing a promising therapeutic agent for severe neutrophilic asthma.

Topics: Animals; Apoptosis; Asthma; Cell Proliferation; HMGB1 Protein; Humans; Hypersensitivity; Inflammation; Inflammation Mediators; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; Muscle, Smooth; Ovalbumin; Proto-Oncogene Proteins c-akt

The role of normal gut microbiota in asthma or ovalbumin (OVA)-induced asthma tolerance (OT) remains unclear. Here, we established mouse models of asthma and OT followed by 2 weeks of antibiotic treatment, to clear the gut microbiota. Antibiotic treatment was found to alleviate allergic asthma accompanied with a reduction of invariant natural killer (iNKT) cells. By RNA-seq analysis, we found that β-adrenergic receptor (ADRB) genes, including

Topics: Animals; Anti-Bacterial Agents; Asthma; Killer Cells, Natural; Mice; Mice, Inbred BALB C; Natural Killer T-Cells; Ovalbumin; Weaning

Asthma is a chronic respiratory disease with symptoms such as expiratory airflow narrowing and airway hyperresponsiveness (AHR). Millions of people suffer from asthma and are at risk of life-threatening conditions. Lactoferrin (LF) is a glycoprotein with multiple physiological functions, including antioxidant, anti-inflammatory, antimicrobial, and antitumoral activities. LF has been shown to function in immunoregulatory activities in ovalbumin (OVA)-induced delayed type hypersensitivity (DTH) in mice. Hence, the purpose of this study was to investigate the roles of LF in AHR and the functions of dendritic cells (DCs) and Th2-related responses in asthma. Twenty 8-week-old male BALB/c mice were divided into normal control (NC), ovalbumin (OVA)-sensitized, and OVA-sensitized with low dose of LF (100 mg/kg) or high dose of LF (300 mg/kg) treatment groups. The mice were challenged by intranasal instillation with 5% OVA on the 21st to 27th day after the start of the sensitization period. The AHR, cytokines in bronchoalveolar lavage fluid, and pulmonary histology of each mouse were measured. Serum OVA-specific IgE and IgG1 and OVA-specific splenocyte responses were further detected. The results showed that LF exhibited protective effects in ameliorating AHR, as well as lung inflammation and damage, in reducing the expression of Th2 cytokines and the secretion of allergen-specific antibodies, in influencing the functions of DCs, and in decreasing the level of Th2 immune responses in a BALB/c mouse model of OVA-induced allergic asthma. Importantly, we demonstrated that LF has practical application in reducing DC-induced Th2 cell responses in asthma. In conclusion, LF exhibits anti-inflammation and immunoregulation activities in OVA-induced allergic asthma. These results suggest that LF may act as a supplement to prevent asthma-induced lung injury and provide an additional agent for reducing asthma severity.

Topics: Animals; Asthma; Cytokines; Dendritic Cells; Lactoferrin; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

This study investigated the effect of Maxing Shigan Decoction(MXSGD) and its disassembled prescriptions against the airway inflammation in respiratory syncytial virus(RSV)-aggravated asthma and the regulation of transient receptor potential vanilloid-1(TRPV1). To be specific, ovalbumin(OVA) and RSV were used to induce aggravated asthma in mice(female, C57BL/6). Then the model mice were intervened by MXSGD and the disassembled prescriptions. The eosinophil(EOS) in peripheral blood, inflammatory cells in bronchoalveolar lavage fluid(BALF), enhanced pause(Penh) variation, and lung pathological damage in each group were observed, and the changes of interleukin(IL)-4, IL-13, substance P(SP), and prostaglandin E2(PGE2) in BALF were mea-sured by enzyme-linked immunosorbent assay(ELISA). Quantitative real time polymerase chain reaction(qPCR) and Western blot were used to detect mRNA and protein of TRPV1 in mouse lung tissue. In the in vitro experiment, 16 HBE cells were stimulated with IL-4 and RSV. Then the changes of TRPV1 expression after the intervention with the serum containing MXSGD and its disassembled prescriptions were observed. Besides, the intracellular Ca~(2+) level after the stimulation with TRPV1 agonist was evaluated. The results showed that the mice in the model group had obvious asthma phenotype, the levels of various inflammatory cells in the peripheral blood and BALF and Penh were significantly increased(P<0.05, P<0.01), and the lung tissue was severely damaged compared with the control group. Compared with the model group, the levels of EOS in the peripheral blood and BALF were significantly decreased in the MXSGD group, the SG group and the MXC group(P<0.05, P<0.01). The levels of WBC and neutrophils in BALF were significantly decreased in the MXSGD group and SG group(P<0.01), the levels of neutrophils in BALF were decreased in the MXC group(P<0.05). The improvement effect of the MXGSD on the level of inflammatory cells in peripheral blood and BALF was better than that of two disassembled groups(P<0.05, P<0.01). After 50 mg·mL~(-1) acetylcholine chloride stimulation, the Penh values of the MXSGD group and the MXC group significantly decreased(P<0.01), and the Penh value of the SG group decreased(P<0.05). The levels of IL-4, IL-13, PGE2 and SP in BALF could be significantly decreased in the MXSGD group(P<0.05, P<0.01), the levels of IL-13 and PGE2 in BALF could be decreased in the MXC group(P<0.05, P<

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Dinoprostone; Disease Models, Animal; Female; Inflammation; Interleukin-13; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Prescriptions; RNA, Messenger; TRPV Cation Channels

Methyl p-coumarate (methyl p-hydroxycinnamate) (MH) is a natural compound found in a variety of plants. In the present study, we evaluated the ameliorative effects of MH on airway inflammation in an experimental model of allergic asthma (AA). In this in vitro study, MH was found to exert anti-inflammatory activity on PMA-stimulated A549 airway epithelial cells by suppressing the secretion of IL-6, IL-8, MCP-1, and ICAM-1. In addition, MH exerted an inhibitory effect not only on NF-κB (p-NF-κB and p-IκB) and AP-1 (p-c-Fos and p-c-Jun) activation but also on A549 cell and EOL-1 cell (eosinophil cell lines) adhesion. In LPS-stimulated RAW264.7 macrophages, MH had an inhibitory effect on TNF-α, IL-1β, IL-6, and MCP-1. The results from in vivo study revealed that the increases in eosinophils/Th2 cytokines/MCP-1 in the bronchoalveolar lavage fluid (BALF) and IgE in the serum of OVA-induced mice with AA were effectively inhibited by MH administration. MH also exerted a reductive effect on the immune cell influx, mucus secretion, and iNOS/COX-2 expression in the lungs of mice with AA. The effects of MH were accompanied by the inactivation of NF-κB. Collectively, the findings of the present study indicated that MH attenuates airway inflammation in mice with AA, suggesting its potential as an adjuvant in asthma therapy.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Epithelial Cells; Inflammation; Interleukin-6; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Topics: Animals; Artemisia; Asthma; Cell Degranulation; Cytokines; Disease Models, Animal; Immunoglobulin G; Inflammation; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Rhinitis, Allergic; Th1 Cells; Th2 Cells; Transcription Factors

The circadian clock is closely associated with inflammatory reactions. Increased inflammatory cytokine levels have been detected in the airways of nocturnal asthma. However, the mechanisms that contribute to the nocturnal increase in inflammatory responses and the relationship with circadian clock remain unknown.. Inflammatory cytokine levels were measured in asthma patients with and without nocturnal symptoms. Allergic airway disease was induced in mice by ovalbumin (OVA), and different periods of light/dark cycles were used to induce circadian rhythm disorders. Serum shock was used to stimulate the rhythmic expression in human bronchial epidermal cells (16HBE). The expression and oscillation of circadian clock genes and inflammatory cytokines in 16HBE cells subjected to brain and muscle ARNT-like protein-1 (BMAL1) and Forkhead Box A2 (FOXA2) knockdown and treatment with a FOXA2 overexpression plasmid were assessed.. Serum IL-6 was found to be significantly higher in asthmatic patients with nocturnal symptoms than those without nocturnal symptoms. The OVA-induced asthma model with a circadian rhythm disorder and 16HBE cells treated with serum shock showed an increase in IL-6 levels and a negative correlation with BMAL1 and FOXA2. The knockdown of BMAL1 resulted in a lower correlation between IL-6 and other rhythm clock genes. Furthermore, knockdown of the BMAL1 and FOXA2 in 16HBE cells reduced the expression and rhythmic fluctuations of IL-6.. Our findings suggest that there are increased IL-6 levels in nocturnal asthma resulting from inhibition of the BMAL1/FOXA2 signalling pathway in airway epithelial cells.

Topics: Animals; Asthma; Cytokines; Hepatocyte Nuclear Factor 3-beta; Humans; Hypersensitivity; Interleukin-6; Mice; Ovalbumin

Global air pollution and diesel exhaust particles (DEPs) generated by intratracheal instillation aggravate asthma. In this study, we evaluated the effect of probiotics via tracheal- or oral-route administration on allergies or asthma. We continuously perfused rats daily, using the oral and tracheal routes, with approximately 10

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; Rats

Airway fibrosis is one of the pathological characteristics of severe asthma. Transforming growth factor (TGF)-β has been known to promote epithelial-mesenchymal transition formation and to play a role in the progression of tissue fibrosis. Cellular communication network factor 2 (CCN2) and fibronectin (FN) are well-known markers of EMT and fibrosis. However, whether AREG is involved in TGF-β-induced CCN2 and FN expression in human lung epithelial cells is unknown.. AREG and FN were analyzed by immunofluorescence staining on ovalbumin-challenged mice. CCN2 and FN expression were evaluated in human lung epithelial (A459) cells following TGF or AREG treatment for the indicated times. Secreted AREG from A549 cells was detected by ELISA. Cell migration was observed by a wound healing assay. Chromatin immunoprecipitation was used to detect the c-Jun binding to the CCN2 promoter.. AREG and FN expression colocalized in lung tissues from mice with ovalbumin-induced asthma by immunofluorescence staining. Moreover, TGF-β caused the release of AREG from A549 cells into the medium. Smad3 siRNA down-regulated AREG expression. AREG also stimulated CCN2 and FN expression, JNK and c-Jun phosphorylation, and cell migration in A549 cells. AREG small interfering (si) RNA inhibited TGF-β-induced expression of CCN2, FN, and cell migration. Furthermore, AREG-induced CCN2 and FN expression were inhibited by EGFR siRNA, a JNK inhibitor (SP600125), and an activator protein-1 (AP-1) inhibitor (curcumin). EGFR siRNA attenuated AREG-induced JNK and c-Jun phosphorylation. Moreover, SP600125 downregulated AREG-induced c-Jun phosphorylation.. These results suggested that AREG mediates the TGF-β-induced EMT in human lung epithelial cells through EGFR/JNK/AP-1 activation. Understanding the role of AREG in the EMT could foster the development of therapeutic strategies for airway remodeling in severe asthma.

Topics: Amphiregulin; Animals; Asthma; Epithelial Cells; Epithelial-Mesenchymal Transition; ErbB Receptors; Fibronectins; Fibrosis; Humans; Lung; Mice; Ovalbumin; RNA, Small Interfering; Transcription Factor AP-1; Transforming Growth Factor beta; Transforming Growth Factor beta1

Asthma is a heterogeneous disorder of the airways related to inflammation; it affects millions of people worldwide. Due to the side effects of inhaled corticosteroids, researchers focused on the therapeutic effects of compounds derived from natural products.. To investigate the therapeutic benefits of Narirutin a valuable flavonoid in Citri Reticulatae Pericarpium for asthma.. Narirutin was extracted using the enzyme-assisted method with the L9 (34) orthogonal array to optimize the temperatures, pH, and reaction time. The mechanism of action of Narirutin was investigated via ELISA, flow cytometry, and Western blot analysis in vivo.. Narirutin suppressed inflammatory cell infiltration in the lung tissue and decreased IgE and IgG1 levels in serum in vivo. It can also alleviate interleukin (IL)-4, IL-5, and interferon-γ concentrations in bronchoalveolar lavage fluid in mice. Moreover, it increased the ratio of CD4+/CD8+ T cells. Additionally, Narirutin significantly suppressed p-ERK1/2 and p-JNK expression in the MAPK signaling pathway.. Narirutin affects the Th1/Th2 imbalance through the p-ERK and p-JNK suppression in the MAPK signaling pathway.

Topics: Animals; Asthma; Disaccharides; Disease Models, Animal; Flavanones; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

The use of ketamine, an anesthetic, as a treatment for asthma has been investigated in numerous studies. However, how ketamine affects asthma is unclear. The present study examined the effects of ketamine on a murine model of mixed-granulocytic asthma, and the role of the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway.. The murine model of mixed-granulocytic asthma was established using ovalbumin (OVA) for sensitization and the combination of OVA and lipopolysaccharides (LPS) for challenge. The main characteristics of asthma, oxidative stress biomarkers, and the expression of the Nrf2 pathway were examined. ML385 was administered to verify the role of the Nrf2 pathway.. Mice in the OVA +LPS group developed asthmatic characteristics, including airway hyperresponsiveness, mixed-granulocytic airway inflammation, mucus overproduction, as well as increased levels of oxidative stress and impaired apoptosis of inflammatory cells. Among the three concentrations, ketamine at 75mg/kg effectively attenuated these asthmatic symptoms, activated the Nrf2 pathway, decreased oxidative stress, and induced apoptosis of eosinophils and neutrophils in bronchoalveolar lavage fluid (BALF) with a reducing level of myeloid cell leukemia 1(Mcl-1). ML385 (an Nrf2 inhibitor) eliminated the protective effects of ketamine on the mixed-granulocytic asthma model.. The study concluded that ketamine reduced oxidative stress and attenuated asthmatic symptoms (neutrophilic airway inflammation) by activating the Nrf2-Keap1 pathway, with 75 mg/kg ketamine showing the best results. Ketamine administration also increased neutrophil and eosinophil apoptosis in BALF, which may contribute to the resolution of inflammation. The use of ketamine as a treatment for asthma may therefore be beneficial.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Kelch-Like ECH-Associated Protein 1; Ketamine; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; Ovalbumin

Bronchial asthma is a chronic inflammatory disease marked by inflammation, oxidative stress, and structural remodeling. Here, we prepared two pomegranate fractions from the seed oil, saponifiable (Sap) and unsaponifiable (UnSap). Two organogels (Orgs) were also formulated with the Sap (Org1) or the UnSap (Org2) fraction and beeswax (BW). All preparations were evaluated in vitro for their antioxidant and anti-inflammatory impacts. The transdermal delivery of the most efficient one was evaluated against ovalbumin (OV)-induced bronchial asthma in rats compared to dexamethasone (DEX). The results showed that the prepared pomegranate fractions and BW had considerable amounts of phenolics (flavonoids and tannins) and triterpenoids. Org1 was shown to be the most effective antioxidant and anti-inflammatory fraction with synergistic activities (combination index, 1), as well as having protective and therapeutic influences on OV-sensitized rats. Org1 inhibited the multiple OV-induced signaling pathways, comprising ROS, WNT/β-catenin, and AKT, with an efficiency superior to DEX. Subsequently, the pro-inflammatory (COX-2, NO, and IL-13), and pro-fibrotic (COL1A1) mediators, oxidative stress, and mucin secretion, were all down-regulated. These outcomes were verified by the histopathological results of lung tissue. Collectively, these outcomes suggest that the transdermal delivery of Org1 to OV-sensitized rats shows promise in the protection and treatment of the pathological hallmarks of asthma.

Topics: Animals; Asthma; Gels; Interleukin-13; Male; Ovalbumin; Oxidative Stress; Plant Oils; Pomegranate; Rats; Seeds; Wnt Signaling Pathway

Allergic asthma was typically considered as an inflammatory disease mediated by type 2 immunity. However, recent studies revealed that asthma is a complex disease displaying a variety of phenotypes and endotypes.. We examined cellular phenotypes in the mouse model of allergic asthma sensitized with different adjuvants. The aim of our study was to determine immunologic cellular characteristics in mouse asthma models induced by ovalbumin (OVA) and a variety of adjuvants.. Mice were sensitized intraperitoneally with the admixture of OVA and various adjuvants such as Alhydrogel (alum), papain, lipopolysaccharide (LPS), or CpG, and subsequently challenged with OVA intranasally. The cells in bronchoalveolar lavage (BAL) fluid, lung, and mediastinal lymph node (mLN) were examined by flow cytometric analyses.. In the lung and BAL fluid, the highest eosinophil levels were observed in the alum group while the highest neutrophil levels were detected in the LPS group. Meanwhile, the LPS group exhibited the most elevated levels of both RORγt+ innate lymphoid cells (ILCs) and IL-17A+ Th cells in the lung and mediastinal lymph node. In the lung, the number of T-bet+ ILCs was highest in the papain group whereas the number of IFN-γ+ Th cells was highest in the CpG group.. Notable variances are found in the composition of immune cells and expression of cytokines at the site of pathogenesis among the different mouse models of allergic asthma created by the sensitization with different adjuvants.

Topics: Adjuvants, Immunologic; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Humans; Immunity, Innate; Inflammation; Lipopolysaccharides; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Papain

To clarify the possible influence of miR-135b on CXCL12 and airway inflammation in children and experimental mice with asthma.. The expressions of miR-135b and CXCL12 were detected using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) in the serum of asthmatic children. Besides, the experimental asthmatic mice were established by aerosol inhalation of ovalbumin (OVA) followed by the treatment with agomiR-135b and antagomir-135b. Pathological changes of lung tissues were observed via HE staining and PAS staining. Besides, the airway hyperresponsiveness of mice was elevated and bronchoalveolar lavage fluid (BALF) was isolated for cell categorization and counting. The inflammatory cytokines in BALF were determined by enzyme-linked immunosorbent assay (ELISA), and the infiltration of Th17 cells in lung tissues was measured using flow cytometry.. MiR-135b was downregulated and CXCL12 was upregulated in asthmatic children and mice. Overexpression of miR-135b may down-regulate CXCL12 expression in the lung of OVA mice, resulting in significant decreases in inflammatory infiltration, hyperplasia of goblet cell, airway hyperresponsiveness, cell quantity, as well as the quantity of eosinophilic granulocytes, neutrophils and lymphocytes in BALF. Also, the levels of inflammatory cytokines (IL-4, IL-5, IL-13 and IL-17) and the ratio of Th17 cells and IL-17 levels in lung tissues were decreased. However, miR-135b downregulation reversed these changes in OVA mice.. MiR-135b may inhibit immune responses of Th17 cells to alleviate airway inflammation and hyperresponsiveness in asthma possibly by targeting CXCL12, showing the potential value in asthma treatment.

Topics: Animals; Asthma; Chemokine CXCL12; Child; Disease Models, Animal; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin

Topics: Animals; Asthma; Becaplermin; Bronchoalveolar Lavage Fluid; Collagen; Flavonoids; Humans; Inflammation; Lung; Mice; MicroRNAs; NF-kappa B; Ovalbumin; Toll-Like Receptor 4

Ruscogenin is a natural product exhibiting anti-inflammatory, antioxidant, and anti-apoptotic effects; however, its effectiveness for asthma management has not yet been reported. The aim of this study was to explore the role of ruscogenin in airway inflammation and apoptosis in asthma.. Ruscogenin reduced oxidative stress and apoptosis in the airway epithelium by inhibiting VDAC1 expression and mitochondrial handling of calcium.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Calcium; Disease Models, Animal; Female; Humans; Hydrogen Peroxide; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Spirostans

As recorded, agarwood has the function of improving qi reception and relieving asthma, but the underlying mechanism is unclear and rarely reported. Therefore, this study explored the anti-asthmatic effect of the alcohol extract of agarwood produced by the whole-tree agarwood-inducing technique(Agar-Wit) in the asthma mouse model induced by intraperitoneal injection of ovalbumin(OVA) + Al(OH)_3 combined with intranasal administration of OVA and the mechanism, and compared the anti-asthmatic effects of agarwood induced with different methods. Firstly, the anti-inflammatory and anti-asthmatic effects of Agar-Wit agarwood in mice were evaluated based on the asthma frequency, lung tissue injury, and peripheral inflammatory white blood cell(WBC) count and eosinophil count. Then, the levels of interleukin-1β(IL-1β), IL-17, and IL-10 in serum of mice were detected by enzyme-linked immunoassay(ELISA) and the expression of inflammation-and apoptosis-related genes in tissues was measured by reverse transcription polyme-rase chain reaction(RT-PCR) so as to preliminarily explore the anti-asthmatic mechanism. RESULTS:: showed that the alcohol extract of Agar-Wit agarwood significantly reduced asthma frequency, relieved pathological injury, improved peripheral WBC count and eosinophil count, decreased the levels of inflammatory cytokines IL-1β and IL-17, elevated the level of anti-inflammatory cytokine IL-10, and down-regulated the mRNA expression of IL-1 R, tumor necrosis factor receptor R(TNFR), nuclear transcription factor-kappa B(NF-κB), Bax, and caspase 3, but had no significant influence on the expression of high-mobility group box 1(HMGB1) protein, caspase 8, and Bcl-2. The effect of Agar-Wit agarwood alcohol extract was better than that of wild agarwood alcohol extract and alcohol extract of agarwood induced with the burning-chisel-drilling method at the same dose. In conclusion, Agar-Wit agarwood can significantly alleviate inflammation and asthma, which is related to its anti-inflammation and anti-apoptosis activity.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Plant Extracts

Asthma is driven by group 2 innate lymphoid cells, antigen-specific CD4+ T helper type 2 cells and their cytokines such as interleukin (IL)-4, IL-5, IL-13. IL-37 is decreased in asthma and negatively related to Th2 cytokines and other pro-inflammatory cytokines. Our study showed that IL-37 level in asthmatic peripheral blood mononuclear cells was lower than in healthy. Further, IL-37 was negatively correlated with exhaled nitric oxide, asthma control test score, atopy and rhinitis history in asthmatics. Then an OVA-induced asthma mice model treated with rhIL-37 was established. An antibody array was employed to uncover altered cytokines induced by IL-37 in mice lung tissue. 20 proteins differentially expressed after rhIL-37 treatment and five of them were validated in asthmatic peripheral blood mononuclear cells. Consistent with cytokine antibody array, CCL3, CCL4, CCL5 decreased after IL-37 administration. While CXCL9 and CXCL13 were no change. We concluded that IL-37 reduce asthmatic symptoms by inhibit pro-inflammatory cytokine such as CCL3, CCL4, CCL5.

Topics: Adult; Animals; Antibodies; Asthma; CD4-Positive T-Lymphocytes; Chemokines, CC; Disease Models, Animal; Female; Humans; Immunity, Innate; Interleukin-1; Leukocytes, Mononuclear; Lung; Male; Mice; Middle Aged; Ovalbumin; Protein Array Analysis; Young Adult

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Lung; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Th17 Cells

Titanium dioxide nanoparticles (TiO

Topics: Animals; Apoptosis; Asthma; bcl-2-Associated X Protein; Bronchoalveolar Lavage Fluid; Carrier Proteins; Caspase 3; Cell Count; Cell Line; Chemical Phenomena; Cytokines; Disease Models, Animal; Gene Expression Regulation; Humans; Hypersensitivity; Immunoglobulin E; Inflammation; Inflammation Mediators; Lung; MAP Kinase Kinase Kinase 5; Mice; Mucus; Nanoparticles; Ovalbumin; Respiratory Hypersensitivity; RNA, Messenger; Thioredoxins; Titanium; Up-Regulation

Asthma is a T helper 2 (Th2) cell-associated chronic inflammatory diseases characterized with airway obstruction, increased mucus production, and eosinophil infiltration. Conventional medications for asthma treatment cannot fully control the symptoms, and potential side effects are also the concerns. Thus, complement or alternative medicine (CAM) became a new option for asthma management. Ding Chuan Tang (DCT) is a traditional Chinese herbal decoction applied mainly for patients with coughing, wheezing, chest tightness, and asthma. Previously, DCT has been proved to improve children airway hyperresponsiveness (AHR) in a randomized and double-blind clinical trial. However, the mechanisms of how DCT alleviates AHR remain unclear. Since asthmatic features such as eosinophil infiltration, IgE production, and mucus accumulation are relative with Th2 responses, we hypothesized that DCT may attenuate asthma symptoms through regulating Th2 cells. Ovalbumin (OVA) was used as a stimulant to sensitize BALB/c mice to establish an asthmatic model. AHR was detected one day before sacrifice. BALF and serum were collected for immune cell counting and antibody analysis. Splenocytes were cultured with OVA in order to determine Th2 cytokine production. Lung tissues were collected for histological and gene expression analyses. Our data reveal that DCT can attenuate AHR and eosinophil accumulation in the 30-day sensitization asthmatic model. Histological results demonstrated that DCT can reduce cell infiltration and mucus production in peribronchial and perivascular site. In OVA-stimulated splenocyte cultures, a significant reduction of IL-5 and IL-13 in DCT-treated mice suggests that DCT may alleviate Th2 responses. In conclusion, the current study demonstrates that DCT has the potential to suppress allergic responses through the reduction of mucus production, eosinophil infiltration, and Th2 activity in asthma.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Down-Regulation; Eosinophils; Female; Immunization; Immunoglobulin E; Interleukin-13; Interleukin-5; Mice, Inbred BALB C; Mucus; Ovalbumin; Plant Extracts; Pneumonia; Spleen

Dimeric translationally controlled tumor protein (dTCTP), also known as histamine-releasing factor, amplifies allergic responses and its production has been shown to increase in inflammatory diseases such as allergic asthma. Despite the critical role of dTCTP in allergic inflammation, little is known about its production pathways, associated cellular networks, and underlying molecular mechanisms. In this study, we explored the dTCTP-mediated inflammatory networks and molecular mechanisms of dTCTP associated with lipopolysaccharides (LPS)-induced severe asthma. LPS stimulation increased dTCTP production by mast cells and dTCTP secretion during degranulation, and extracellular dTCTP subsequently increased the production of pro-inflammatory molecules, including IL-8, by airway epithelial cells without affecting mast cell activation. Furthermore, dimeric TCTP-binding peptide 2 (dTBP2), a dTCTP inhibitor peptide, selectively blocked the dTCTP-mediated signaling network from mast cells to epithelial cells and decreased IL-8 production through IkB induction and nuclear p65 export in airway epithelial cells. More importantly, dTBP2 efficiently attenuated LPS-induced severe airway inflammation in vivo, resulting in decreased immune cell infiltration and IL-17 production and attenuated dTCTP secretion. These results suggest that dTCTP produced by mast cells exacerbates airway inflammation through activation of airway epithelial cells in a paracrine signaling manner, and that dTBP2 is beneficial in the treatment of severe airway inflammation by blocking the dTCTP-mediated inflammatory cellular network.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Coculture Techniques; Cytokines; Disease Models, Animal; Epithelial Cells; HEK293 Cells; Humans; Inflammation Mediators; Lipopolysaccharides; Lung; Male; Mast Cells; Mice, Inbred C57BL; Ovalbumin; Paracrine Communication; Peptides; Pneumonia; Severity of Illness Index; Signal Transduction; Transcription Factor RelA; Tumor Protein, Translationally-Controlled 1

It is becoming increasingly clear that environment factors during early life play a pivotal role in the development of allergic asthma. Among these, a traditional farm is one of the strongest protective environments, and the protective effects have been, at least in part, attributed to the high-level exposure to lipopolysaccharide (LPS) on farms. However, the underlying mechanisms remain elusive, especially in ovalbumin (OVA)-induced neonatal allergic asthma model. Here, we used the OVA-induced asthma model in two age groups, neonatal and adult, when mice were first sensitized with peritoneal OVA/alum as neonates and adults, respectively. LPS was injected in the peritoneal cavity before OVA/alum sensitization. The effects of LPS treatment on allergic airway inflammation in the lung and the immune milieu in the peritoneal cavity were determined and compared between these two age groups. We found that LPS treatment abrogated the development of Th2 allergic airway responses in the neonatal group. In the adult group, the ameliorated Th2 allergic responses were accompanied with Th17 responses and neutrophil infiltration upon LPS treatment. We further investigated the immune milieu in the peritoneal cavity to elucidate the underlying mechanisms of this age-dependent difference. Our data show that in neonatal mice, LPS treatment significantly reduced the number of inflammatory monocytes in the peritoneal cavity. In the adult group, LPS treatment shifted the function of these cells which associated with Th1 and Th17 polarization. Our results provide more evidence that immunity in early life is distinct from that in adults, especially in the peritoneal cavity, and emphasize the importance of timing for the intervention of allergic asthma. Our results suggest that LPS treatment during early life is protective for the development of Th2 allergic responses. On the other hand, it might lead to a more severe phenotype of asthma when dampening the Th2 responses in adult mice.

Topics: Age Factors; Alum Compounds; Animals; Animals, Newborn; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Inflammation; Lipopolysaccharides; Lung; Male; Mice; Mice, Inbred BALB C; Monocytes; Ovalbumin; Th17 Cells; Th2 Cells

To investigate the therapeutic effect of LiuJunZi decoction (LJZD) in an experimental model of asthma and uncover its potential mechanism.. The ovalbumin (OVA) was applied to induce asthma in Balb/C mice, and LJZD was orally administrated to asthmatic mice. The lung function and histological lesion were evaluated by airway hyperresponsiveness assay, lung edema assay, and hematoxylin and eosin staining. The amounts of CD4. LJZD improves OVA-induced asthma in Balb/C mice, which is manifested by decreasing lung edema, Penh levels, lung histological lesion, and inflammatory cell infiltration. LJZD increased the number of CD4. LJZD improved OVA-induced asthma in Balb/C mice by inhibiting allergic inflammation and Th2 immunoreaction, which might be associated with the inactivation of the NF-

Topics: Animals; Anti-Asthmatic Agents; Asthma; Disease Models, Animal; Drugs, Chinese Herbal; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; T-Lymphocytes, Regulatory; Th2 Cells

Asthma animal models provide valuable information about the pathogenesis and the treatment of asthma. An ovalbumin (OVA)/complete Freund's adjuvant (CFA)-sensitized model was developed to induce neutrophil-dominant asthma and to investigate whether fungal immunomodulatory peptide-fve (FIP-fve) could improve asthma features in the OVA/CFA-sensitized model.. We used female BALB/c mice and sensitized them intraperitoneally with OVA/CFA on days 1, 2, and 3. On days 14, 17, 21, 24, and 27, they were challenged with intranasal OVA. The airway hyper-responsiveness (AHR) was detected by BUXCO, inflammatory cells were stained with Liu's stain, the cytokines were detected using ELISA, and the airway inflammation was analyzed with hematoxylin and eosin stain.. According to the results, OVA/CFA sensitization could induce AHR, high levels of IgE, and inflammatory cells especially neutrophils infiltration in the lung and airway inflammation. IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, IL-17, IL-25, IL-33, and transforming growth factor-β (TGF-β) increased in the OVA/CFA-sensitized mice. OVA/CFA-sensitized mice treated with FIP-fve not only increased IL-12 and IFN-γ but also decreased IL-4, IL-5, IL-6, IL-8, IL-13, IL-17, IL-25, IL-33, and TGF-β in the bronchoalveolar lavage fluid. Moreover, FIP-fve significantly decreased neutrophil infiltration in the lung.. The OVA/CFA model induced neutrophilic asthma successfully, and FIP-fve improved neutrophil-dominant asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Biomarkers; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Freund's Adjuvant; Fungal Proteins; Immunoglobulin E; Mice; Mice, Inbred BALB C; Neutrophil Infiltration; Neutrophils; Ovalbumin; Real-Time Polymerase Chain Reaction; Treatment Outcome

Allergic asthma is a complicated respiratory problem characterized by airway inflammation, airway hyperresponsiveness (AHR), breathlessness, mucus hyper-secretion, and goblet cell hyperplasia. Asthma is controlled by genetic and environmental factors. Allergy is the main trigger of asthma and is mediated by Th2 cytokines along with IgE production. Vitamin D (Vit D) is the main supplementary factor for the immune system. In the present study, we investigated the effect of Vit D on the exacerbation of allergic asthma. A murine model of allergic asthma was induced by ovalbumin (OVA) in four of five groups of studied female BALB/c mice (each group, n=20). One group was considered as control. Of OVA-induced mice, two groups received Vit D via oral (10,000 IU/kg diet) or intranasal (inhalation) forms (30 min on days 25, 27, and 29), and the third group received budesonide. At least, AHR, the levels of IL-4, IL-5, IL-13, and INF-g in bronchoalveolar lavage fluid (BALF), serum IgE and histamine, IL-25 and IL-33 gene expression, as well as histopathology study of the lung were done. The Penh values, type2 Cytokines in BALF (in both protein and molecular levels), total IgE and histamine, perivascular and peribronchial inflammation, goblet cell hyperplasia, and mucus hypersecretion decreased significantly in both oral and intranasal Vit D-treated asthmatic mice groups, especially on day 38 of orally treated mice. Here, we found Vit D as a promising agent in control of allergic asthma with a remarkable ability to decrease the severity of inflammation. Therefore, Vit D sufficiency is highly recommended in asthmatic patients.

Topics: Animals; Asthma; Biomarkers; Bronchi; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Disease Susceptibility; Immunoglobulin E; Mice; Ovalbumin; Vitamin D

A 7-amino acid peptide (7P), (Gly-Gln-Thr-Tyr-Thr-Ser-Gly) is one of the synthesized mimic polypeptides, which is the second envelope protein at hypervariable region 1 of chronic hepatitis C virus (HCV HVR1). It contributed to the anti-inflammatory reaction and inhibited lung Th9 responses in asthma through binding to CD81. In this study, we examined the effects of 7P on bronchoconstriction, acute inflammation of the airways, and lung Th2-type responses during allergic lung inflammation. Our results determined that 7P decreased bronchoconstriction and inhibited both acute inflammatory cytokines (TNFα, IL-1β, and IL-6) and Th2 cell cytokine responses (IL-5, IL-4, and IL-13) during allergic lung inflammation. 7P directly inhibited lung Th2 cell differentiation (7P: 5.1% vs. vehicle:12.2% and control 7P:12.2%) and suppressed airway inflammatory cytokine signal transduction to decrease Th2 cell response. Overall, 7P significantly decreased airway hyperresponsiveness (AHR), airway inflammation, and Th2 responses, which may serve as a novel therapeutic candidate during allergic lung inflammation.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Cell Differentiation; Cytokines; Disease Models, Animal; Inflammation; Male; Mice, Inbred C57BL; Ovalbumin; Peptides; Respiratory Hypersensitivity; Th2 Cells

Accumulating evidence has implicated the potential of natural compounds in treatment of asthma. Bixin is a natural food coloring isolated from the seeds of Bixa Orellana, which possesses anti-tumor, anti-inflammatory and antioxidative properties. Nevertheless, its therapeutic effect in asthma has not been elucidated. Our present study demonstrated that administration of Bixin suppressed allergic airway inflammation and reversed glucocorticoids resistance, as well as alleviated airway remodeling and airway hyperresponsiveness (AHR) in asthmatic mice. In vitro studies showed that Bixin treatment could inhibit the development of epithelial-mesenchymal transition (EMT) mediated by transforming growth factor beta (TGF-β) signaling. Importantly, Bixin antagonized activation of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway both in vitro and in vivo. Above all, our findings reveal that Bixin functions as a potent antagonist of PI3K/Akt signaling to protect against allergic asthma, highlighting a novel strategy for asthma treatment based on natural products.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Asthma; Bixaceae; Carotenoids; Disease Models, Animal; Female; Humans; Mice; Mice, Inbred BALB C; Oncogene Protein v-akt; Ovalbumin; Phosphatidylinositol 3-Kinases; Respiratory Hypersensitivity; Signal Transduction; Transforming Growth Factor beta

The fermented soy product ImmuBalance contains many active ingredients and its beneficial effects on some allergic diseases have been reported. We hypothesized that ImmuBalance could have potential effects on airway inflammation in a murine model of asthma. Mice sensitized and challenged with ovalbumin developed airway inflammation. Bronchoalveolar lavage fluid was assessed for inflammatory cell counts and levels of cytokines. Lung tissues were examined for cell infiltration and mucus hypersecretion. Oral administration of ImmuBalance significantly inhibited ovalbumin-induced eosinophilic inflammation and decreased Th2 cytokine levels in bronchoalveolar lavage fluid (

Topics: Animals; Asthma; Body Weight; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Diet; Disease Models, Animal; Eosinophils; Feeding Behavior; Female; Fermented Foods; Glycine max; Immunoglobulin E; Inflammation; Lung; Mice, Inbred BALB C; Ovalbumin

Allergic asthma is well known as a common respiratory disorder comprising an allergic inflammatory nature and excessive immune characteristic.

Topics: Adenosine; Allergens; Animals; Asthma; Disease Models, Animal; DNA Methylation; Epigenesis, Genetic; Epigenome; Female; Gene Expression Profiling; Humans; Hypersensitivity; Immunity; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Signal Transduction

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Brucea javanica; Cytokines; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Quassins; Receptors, Notch; Th1-Th2 Balance

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Asthma; Cell Line; Cell Movement; Cell Proliferation; Disease Models, Animal; Epithelial Cells; Epithelial-Mesenchymal Transition; Female; Humans; Interleukin-4; Lung; Mice, Inbred BALB C; Morphinans; Ovalbumin; Signal Transduction; Smad3 Protein; Transforming Growth Factor beta1

Allergic asthma is a chronic airway inflammatory disease with a number of cytokines participating in its pathogenesis and progress. Interleukin (IL)-22, which is derived from lymphocytes, acts on epithelial cells and play a role in the chronic airway inflammation. However, the actual role of IL-22 in allergic asthma is still unclear. Therefore, we explored the effect of IL-22 on allergic airway inflammation and airway hyperresponsiveness (AHR) in an ovalbumin (OVA)-induced asthma mouse model.. To evaluate the effect of IL-22 in an allergic asthma model, BALB/c mice were sensitized and challenged with OVA; then the recombinant mouse IL-22 was administered intranasally 24 h prior to each challenge. The IL-22 levels in lung homogenates and bronchoalveolar lavage fluid (BALF) were measured by enzyme linked immunosorbent assay, respectively. AHR was evaluated through indicators including airways resistance (Rrs), elastance (Ers) and compliance (Crs); the inflammatory cell infiltration was assessed by quantification of differential cells counts in BALF and lung tissues stained by hematoxylin and eosin (H&E); IL-22 specific receptors were determined by immunohistochemistry staining.. The concentration of IL-22 was significantly elevated in the OVA-induced mice compared with the control mice in lung homogenates and BALF. In the OVA-induced mouse model, IL-22 administration could significantly attenuate AHR, including Rrs, Ers and Crs, decrease the proportion of eosinophils in BALF and reduce inflammatory cell infiltration around bronchi and their concomitant vessels, compared with the OVA-induced group. In addition, the expression of IL-22RA1 and IL-10RB in the lung tissues of OVA-induced mice was significantly increased compared with the control mice, while it was dramatically decreased after the treatment with IL-22, but not completely attenuated in the IL-22-treated mice when compared with the control mice.. Interleukin-22 could play a protective role in an OVA-induced asthma model, by suppressing the inflammatory cell infiltration around bronchi and their concomitant vessels and airway hyperresponsiveness, which might associate with the expression of its heterodimer receptors. Thus, IL-22 administration might be an effective strategy to attenuate allergic airway inflammation.

Topics: Animals; Asthma; Disease Models, Animal; Female; Interleukin-22; Interleukins; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Topics: Airway Remodeling; Animals; Asthma; Bronchial Arteries; Disease Models, Animal; Endothelial Cells; Endothelial Growth Factors; Humans; Mice; Mice, Inbred BALB C; Ovalbumin; Peptides, Cyclic; Vascular Endothelial Growth Factor A

Neutrophil cytosolic factor 1 (

Topics: Animals; Asthma; Disease Models, Animal; Eosinophils; Female; Humans; Immunity, Innate; Lung; Lymphocyte Activation; Mice; Mice, Transgenic; Mutation; NADPH Oxidases; Ovalbumin; Reactive Oxygen Species; Signal Transduction; Th1 Cells

Topics: Airway Remodeling; Animals; Asthma; Drug Resistance; Hyaluronan Synthases; Mice; Mice, Knockout; Ovalbumin; Steroids

It is largely known that photobiomodulation (PBM) has beneficial effects on allergic pulmonary inflammation. Our previous study showed an anti-inflammatory effect of the PBM in an acute experimental model of asthma, and we see that this mechanism is partly dependent on IL-10. However, it remains unclear whether the activation of regulatory T cells is mediated by PBM in a chronic experimental model of asthma. In this sense, the objective of this study was to verify the anti-inflammatory role of the PBM in the pulmonary inflammatory response in a chronic experimental asthma model. The protocol used for asthma induction was the administration of OVA subcutaneously (days 0 and 14) and intranasally (3 times/week, for 5 weeks). On day 50, the animals were sacrificed for the evaluation of the different parameters. The PBM used was the diode, with a wavelength of 660 nm, a power of 100 mW, and 5 J for 50 s/point, in three different application points. Our results showed that PBM decreases macrophages, neutrophils, and lymphocytes in the bronchoalveolar lavage fluid (BALF). Moreover, PBM decreased the release of cytokines by the lung, mucus, and collagen in the airways and pulmonary mechanics. When we analyzed the percentage of Treg cells in the group irradiated with laser, we verified an increase in these cells, as well as the release of IL-10 in the BALF. Therefore, we conclude that the use of PBM therapy in chronic airway inflammation attenuated the inflammatory process, as well as the pulmonary functional and structural parameters, probably due to an increase in Treg cells.

Topics: Animals; Anti-Inflammatory Agents; Asthma; CD4-Positive T-Lymphocytes; Disease Models, Animal; Forkhead Transcription Factors; Inflammation; Interleukin-10; Low-Level Light Therapy; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Asthma is an inflammatory airway disease affecting most of the population in the world. The current medication for asthma relieves airway inflammation but it has serious adverse effects. Biochanin A (BCA), a phytoestrogen, is an active component present in red clover, alfalfa, soy having anti-oxidant and anti-inflammatory properties. BCA was identified as a natural activator of peroxisome proliferator-activated receptor-gamma (PPARγ).. The study aims to evaluate the effects of BCA in ovalbumin (OVA)-induced murine model of asthma and to study the role of PPARγ.. We found that BCA administration reduced the severity of murine allergic asthma as evidenced histologically, and measurement of allergen-specific IgE levels in serum as well as in BAL fluid. BCA also reversed the elevated levels of inflammatory cytokines, cell infiltration, protein leakage into the airways and expression of hemoxygenase-1 in OVA-induced lungs. Further, we confirmed that BCA mediated inhibitory effects are mediated through PPARγ as assessed by treatment with PPARγ antagonist GW9662.. Our results suggest that BCA is efficacious in a preclinical model of asthma and may have the potential for the treatment of asthma in humans.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Disease Models, Animal; Female; Genistein; Inflammation; Mice; Ovalbumin; PPAR gamma; Respiratory Tract Diseases; Signal Transduction; Treatment Outcome

Asthma characterized by airway hyperresponsiveness, inflammation, fibrosis, and angiogenesis. SRY-related HMG-box 18 (SOX18) is an important transcription factor involved in angiogenesis, tissue injury, wound-healing, and in embryonic cardiovascular and lymphatic vessels development. The role of angiogenic transcription factors, SOX18 and the related, prospero homeobox 1 (PROX1) and chicken ovalbumin upstream promoter transcription factor II (COUP-TFII), in asthma has had limited study.. In this study, we aimed to elucidate the role of SOX18 in the pathogenesis of bronchial asthma.. Plasma SOX18 protein was measured in control subjects, and subject with stable or exacerbated asthma. SOX18, PROX1, and COUP-TFII protein was measured by western blot, and immunohistochemistry in a murine model of ovalbumin-induced allergic asthma (OVA). SOX18, PROX1, and COUP-TFII protein was measured in lung human microvascular endothelial cells (HMVEC-L) and normal human bronchial epithelial (NHBE) cells treated with house dust mite (. Plasma SOX18 tended to be higher in subject with asthma compared to control subjects and increased more during exacerbation as compared to stable disease. In mice, OVA challenge lead to increased lung SOX18, PROX1, COUP-TFII, mucous gland hyperplasia and submucosal collagen. In NHBE cells, SOX18, PROX1 and COUP-TFII increased following. These results suggest that SOX18 may be involved in asthma pathogenesis and be associated with asthma exacerbation.

Topics: Adult; Aged; Allergens; Animals; Antigens, Dermatophagoides; Arthropod Proteins; Asthma; Bronchoalveolar Lavage Fluid; Cell Line; COUP Transcription Factor II; Cysteine Endopeptidases; Disease Progression; Female; Fibrosis; Homeodomain Proteins; Humans; Interleukin-5; Lung; Male; Mice, Inbred BALB C; Middle Aged; Neovascularization, Physiologic; Ovalbumin; SOXF Transcription Factors; Tumor Suppressor Proteins; Vascular Endothelial Growth Factor A

Although the importance of mast cells in asthma has been studied, mast cellsinduced global changes in lungs are largely unknown. Data-driven identification contributes to discovering significant biomarkers or therapeutic targets, which are the basis of effective clinical medications.. This study aims to explore the effects of mast cells on gene expression in asthmatic lungs, and to assess the curative effects of inhaled budesonide (BUD).. Pulmonary gene expression in KitWsh mice with or without mast cell engraftment was analyzed with R software. Functional enrichment of Gene Ontology and KEGG was carried out through the DAVID online tool. Hub genes were identified with String and Cytoscape software.. The array analyses showed that the mast cell engraftment enhanced inflammation/immune response, cytokine/chemokine signal, and monocyte/neutrophil/lymphocyte chemotaxis. Interleukin (IL)-6 was identified to be a significant hub gene with the highest interaction degree. Based on this, the effects of BUD were investigated on the aspects of anti-inflammation. BUD's treatment was found to reduce serum IL-6 content and pulmonary inflammation in ovalbumin-induced asthma rats. The treatment also downregulated beta-tryptase expression both in lung tissues and serum. Morphologically, the accumulation and degranulation of mast cells were significantly suppressed. Notably, the effects of BUD on inflammation and degranulation were comparable with Tranilast (a classic mast cell inhibitor), while a remarkable synergy was not observed.. This study presented a unique pulmonary gene profile induced by mast cell engraftment, which could be reversed through blockage of mast cells or inhaled BUD.

Topics: Administration, Inhalation; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Budesonide; Data Analysis; Drug Delivery Systems; Gene Expression Regulation; Lung; Male; Mast Cells; Mice; Ovalbumin; Rats; Rats, Sprague-Dawley; Treatment Outcome

Group 2 innate lymphoid cells (ILC2s) are known to serve important functions in the pathogenesis of allergic airway inflammation. Studies have shown that acupuncture has an anti-inflammatory effect in the airways. However, how acupuncture treatment affects innate immunity, especially with regard to the function of ILC2s in ovalbumin (OVA)-induced allergic airway inflammation, is poorly understood.. BALB/c mice were injected and subsequently challenged with OVA ± treated with manual acupuncture. At the end of the experimental course, lung function was assessed by measurement of airway resistance (R. The results showed that airway inflammation and mucus secretion were significantly suppressed by acupuncture treatment. R. In conclusion, this study has demonstrated that acupuncture treatment alleviates allergic airway inflammation and inhibits pulmonary ILC2 influx and IL-5, IL-9 and IL-13 production. The inhibition of ILC2s by acupuncture may be associated with the IL-33/ST2-signaling pathway and IL-25 levels, thereby offering protection from the respiratory inflammation associated with asthma.

Topics: Acupuncture Therapy; Animals; Asthma; Bronchoalveolar Lavage Fluid; Female; Humans; Immunity, Innate; Interleukin-1 Receptor-Like 1 Protein; Interleukins; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin

Allergen-specific immunotherapy via the skin targets a tissue rich in antigen-presenting cells, but can be associated with local and systemic side effects. Allergen-polysaccharide neoglycogonjugates increase immunization efficacy by targeting and activating dendritic cells via C-type lectin receptors and reduce side effects.. We investigated the immunogenicity, allergenicity, and therapeutic efficacy of laminarin-ovalbumin neoglycoconjugates (LamOVA).. The biological activity of LamOVA was characterized in vitro using bone marrow-derived dendritic cells. Immunogenicity and therapeutic efficacy were analyzed in BALB/c mice. Epicutaneous immunotherapy (EPIT) was performed using fractional infrared laser ablation to generate micropores in the skin, and the effects of LamOVA on blocking IgG, IgE, cellular composition of BAL, lung, and spleen, lung function, and T-cell polarization were assessed.. Conjugation of laminarin to ovalbumin reduced its IgE binding capacity fivefold and increased its immunogenicity threefold in terms of IgG generation. EPIT with LamOVA induced significantly higher IgG levels than OVA, matching the levels induced by s.c. injection of OVA/alum (SCIT). EPIT was equally effective as SCIT in terms of blocking IgG induction and suppression of lung inflammation and airway hyperresponsiveness, but SCIT was associated with higher levels of therapy-induced IgE and TH2 cytokines. EPIT with LamOVA induced significantly lower local skin reactions during therapy compared to unconjugated OVA.. Conjugation of ovalbumin to laminarin increased its immunogenicity while at the same time reducing local side effects. LamOVA EPIT via laser-generated micropores is safe and equally effective compared to SCIT with alum, without the need for adjuvant.

Topics: Allergens; Animals; Asthma; beta-Glucans; Lasers; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia

When the integrity of airway epithelium is destroyed, the ordered airway barrier no longer exists and increases sensitivity to viral infections and allergens, leading to the occurrence of airway inflammation such as asthma. Here, we found that galectin-7 transgenic(+) mice exhibited abnormal airway structures as embryos and after birth. These abnormalities included absent or substantially reduced pseudostratified columnar ciliated epithelium and increased monolayer cells with irregular arrangement and widening of intercellular spaces. Moreover, airway tissue from galectin-7 transgenic(+) mice showed evidence of impaired cell-cell junctions and decreased expression of zonula occludens-1(ZO-1) and E-cadherin. When treated with respiratory syncytial virus (RSV) or ovalbumin (OVA), galectin-7 transgenic(+) mice developed substantially increased bronchial epithelial detachment and apoptosis, airway smooth muscle and basement membrane thickening, and enhanced airway responsiveness. We found that Galectin-7 localized in the cytoplasm and nucleus of bronchial epithelial cells, and that increased apoptosis was mediated through mitochondrial release of cytochrome c and upregulated JNK1 activation and expression of caspase-3 in galectin-7 Tg(+) mice. These findings suggested that Galectin-7 causes airway structural defects and destroys airway epithelium barrier, which predispose the airways to RSV or OVA-induced epithelial apoptosis, injury, and other asthma responses.

Topics: Animals; Apoptosis; Asthma; Bronchi; Cadherins; Disease Models, Animal; Epithelial Cells; Galectins; Mice, Transgenic; Ovalbumin; Rats, Sprague-Dawley; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses; Tight Junctions

Daphne pseudomezereum var. koreana Hamaya is distributed in the Gangwon-do of South Korea and is traditionally used to treat chronic inflammatory diseases, including rheumatoid arthritis.. We investigated the anti-inflammatory effect of biflavonoid-rich fraction (BF) obtained from an extract of D. pseudomezereum leaves on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and mouse model of ovalbumin (OVA)-induced allergic asthma.. Neochamaejasmin B (NB) and chamaejasmin D (CD) were spectroscopically characterized as major components of BF obtained from the leaves of D. pseudomezereum. RAW264.7 cells pretreated with NB, CD and BF and activated by LPS (500 ng/ml) were used to assess the anti-inflammatory effects of these materials in vitro. To evaluate the protective effect of BF on allergic asthma, female BALB/c mice were sensitized to OVA by intraperitoneal (i.p.) injection and treated with BF by oral administration (15 or 30 mg/kg).. Pretreatment with BF inhibited LPS-stimulated nitric oxide (NO), TNF-α and IL-6, and led to upregulation of heme oxygenase-1 (HO-1) in RAW264.7 macrophages. Orally administered BF significantly inhibited the recruitment of eosinophils and the production of IL-5, IL-6, IL-13 and MCP-1 as judged by the analysis of BALF from OVA-induced asthma animal model. BF also decreased the levels of IgE in the serum of asthmatic mice. BF suppressed the influx of inflammatory cells into nearby airways and the hypersecretion of mucus by the airway epithelium of asthmatic mice. In addition, the increase in Penh in asthmatic mice was reduced by BF administration. Furthermore, BF led to Nrf2 activation and HO-1 induction in the lungs of mice.. These data have shown the anti-asthmatic effects of BF, and therefore we expect that BF may be a potential candidate as a natural drug/nutraceutical for the prevention and treatment of allergic asthma.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Biflavonoids; Daphne; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Inflammation; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; RAW 264.7 Cells

Loki Zupa (LKZP) decoction is one of the herbal prescriptions in traditional Uyghur medicine, which is commonly used for treating airway abnormality. However, underlying pathological mechanism and pathways involved has not been well studied.. In this paper, we aim to further confirmed the anti-inflammatory and anti-fibrotic role of LKZP decoction in airway, and uncover the passible mechanism involved via comprehensive quantitative proteomic DIA-MS analysis.. Mice asthmatic model was established with sensitizing and challenging with OVA. Lung function, pathological status, and inflammatory cytokines were assessed. Total of nine lung tissues were analyzed using proteomic DIA-MS analysis and 18 lung tissues were subjected to PRM validation.. Total of 704 differentially expressed proteins (DEPs) (363 up regulated, 341 down regulated) were quantified in comparison of asthmatic and healthy mice, while 152 DEPs (91 up regulated, 61 down regulated) were quantified in LKZP decoction treated compared to asthmatic mice. Total of 21 proteins were overlapped between three groups. ECM-receptor interaction was significantly enriched and commonly shared between downregulated DEPs in asthma and upregulated DEPs in LKZP decoction treated mice. Total of 20 proteins were subjected to parallel reaction monitoring (PRM) analysis and 16 of which were quantified. At last, two proteins, RMB 10 and COL6A6, were validated with significant difference (P < 0.001) in protein abundance.. Our results suggest that attenuated airway inflammation and fibrosis caused by LKZP decoction may associated with ECM-receptor interaction and RMB 10 and COL6A6 may be targeted by LKZP decoction in OVA-induced asthmatic mice.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Female; Inflammation; Medicine, Chinese Traditional; Mice; Mice, Inbred BALB C; Ovalbumin; Proteomics; Receptors, Cell Surface

Recent studies have revealed that YKL-40 is involved in the pathogenesis of asthma. However, its specific mechanism remains unclear. The present study aims to investigate the effect of adenovirus vector-mediated YKL-40 short hairpin RNA (shRNA) on regulation of airway inflammation in a murine asthmatic model. Mice were assessed for airway hyperresponsiveness (AHR), total leukocytes and the percentage of eosinophil cells in bronchoalveolar lavage fluid (BALF). YKL-40 mRNA and protein expression levels were detected using quantitative real-time PCR and western blot assays. Enzyme-linked immunosorbent assay (ELISA) was used to detect YKL-40 and eosinophil-related chemokine expression levels in BALF and serum. Lung histology analyses were performed to evaluate the degree of inflammatory cell infiltration around the airway and airway mucus secretion.YKL-40 shRNA significantly inhibited the YKL-40 gene expression in asthmatic mice. In addition, YKL-40 shRNA alleviated eosinophilic airway inflammation, AHR, airway mucus secretion and decreased the levels of YKL-40 in BALF and serum in a murine asthmatic model. The levels and mRNA expression of IL-5, IL-13 in asthmatic mice lung tissues, eotaxin, and GM-CSF in BALF and serum significantly decreased. Bone marrow signaling molecules including IL-5, eotaxin, and GM-CSF were correlated with decreased levels of YKL-40. The study reveals that YKL-40 could be involved in asthma inflammation by altering bone marrow signaling molecules. YKL-40 gene RNA interference could provide new therapeutic strategies for asthma.

Topics: Adenoviridae; Animals; Asthma; Chitinase-3-Like Protein 1; Disease Models, Animal; Eosinophils; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Small Interfering

Asthma is an airway inflammatory disease and a major health problem worldwide. Anti-inflammatory steroids and bronchodilators are the gold-standard therapy for asthma. However, they do not prevent the development of the disease, and critically, a subset of asthmatics are resistant to steroid therapy.. To elucidate the therapeutic potential of human β-defensins (hBD), such as hBD2 mild to moderate and severe asthma.. We investigated the role of hBD2 in a steroid-sensitive, house dust mite-induced allergic airways disease (AAD) model and a steroid-insensitive model combining ovalbumin-induced AAD with C muridarum (Cmu) respiratory infection.. In both models, we demonstrated that therapeutic intranasal application of hBD2 significantly reduced the influx of inflammatory cells into the bronchoalveolar lavage fluid. Furthermore, key type 2 asthma-related cytokines IL-9 and IL-13, as well as additional immunomodulating cytokines, were significantly decreased after administration of hBD2 in the steroid-sensitive model. The suppression of inflammation was associated with improvements in airway physiology and treatment also suppressed airway hyper-responsiveness (AHR) in terms of airway resistance and compliance to methacholine challenge.. These data indicate that hBD2 reduces the hallmark features and has potential as a new therapeutic agent in allergic and especially steroid-resistant asthma.

Topics: Airway Resistance; Animals; Asthma; beta-Defensins; Bronchoalveolar Lavage Fluid; Chlamydia Infections; Chlamydia muridarum; Disease Models, Animal; Inflammation; Interleukin-13; Interleukin-9; Lung; Lung Compliance; Mice; Ovalbumin; Pyroglyphidae; Respiratory Hypersensitivity; Respiratory Tract Infections

Allergic asthma is a chronic inflammatory lung disease characterized by a Th2-type immune response pattern. The development of nonspecific immunotherapy is one of the primary goals for the control of this disease.. In this study, we evaluated the therapeutic effects of Lactococcus lactis-producing mycobacterial heat shock protein 65 (LLHsp65) in an ovalbumin (OVA)-induced allergic asthma model. OVA-challenged BALB/c mice were orally administrated with LLHsp65 for 10 consecutive days. The results demonstrate that LLhsp65 attenuates critical features of allergic inflammation, like airway hyperresponsiveness and mucus production. Likewise, the treatment decreases the pulmonary eosinophilia and the serum level of OVA-specific IgE. In addition to deviating immune responses towards Th1-cytokine profile, increase regulatory T cells, and cytokine levels, such as IL-6 and IL-10.. Our results reveal that the mucosal immunotherapy of LLHsp65 significantly reduces the overall burden of airway allergic inflammation, suggesting a promising therapeutic strategy for allergic asthma treatment.. This research reveals new perspectives on nonspecific immunotherapy based on the delivery of recombinant proteins by lactic acid bacteria to treat of allergic disorders.

Topics: Administration, Oral; Animals; Asthma; Bacterial Proteins; Bronchoalveolar Lavage Fluid; Chaperonin 60; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Immunotherapy; Inflammation; Lactococcus lactis; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory

Callicarpa japonica Thunb., as an herbal medicine has been used for the treatment of inflammatory diseases in China and Korea.. Ultra performance liquid chromatography-photodiode array-quadrupole time-of-flight mass spectrometer (UPLC-PDA-QTof MS) was used to detect the major phenylethanoid glycosides in the C. japonica extract. BALB/c mice were intraperitoneally sensitized by ovalbumin (OVA) (on days 0 and 7) and challenged by OVA aerosol (on days 11-13) to induce airway inflammatory response. The mice were also administered with C. japonica Thunb. (CJT) (20 and 40 mg/kg Per oral) on days 9-13. CJT pretreatment was conducted in lipopolysaccharide (LPS)-stimulated RAW264.7 or phorbol 12-myristate 13-acetate (PMA)-stimulated A549 cells.. CJT administration significantly reduced the secretion of Th2 cytokines, TNF-α, IL-6, immunoglobulin E (IgE) and histamine, and the recruitment of eosinophils in an OVA-exposed mice. In histological analyses, the amelioration of inflammatory cell influx and mucus secretion were observed with CJT. The OVA-induced airway hyperresponsiveness (AHR), iNOS expression and NF-κB activation were effectively suppressed by CJT administration. In addition, CJT led to the upregulation of HO-1 expression. In an in vitro study, CJT pretreatment suppressed the LPS-induced TNF-α secretion in RAW264.7 cells and attenuated the PMA-induced IL-6, IL-8 and MCP-1 secretion in A549 cells. These effects were accompanied by downregulated NF-κB phosphorylation and by upregulated HO-1 expression.. These results suggested that CJT has protective activity against OVA-induced airway inflammation via downregulation of NF-κB activation and upregulation of HO-1, suggesting that CJT has preventive potential for the development of allergic asthma.

Topics: A549 Cells; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchial Hyperreactivity; Bronchoconstriction; Callicarpa; Cytokines; Disease Models, Animal; Female; Heme Oxygenase-1; Humans; Lung; Membrane Proteins; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Plant Extracts; RAW 264.7 Cells; Signal Transduction; Up-Regulation

Bronchial asthma (BA) is a chronic disease of the airways. The great majority of BA exacerbations are associated with respiratory viral infections. Recent findings point out a possible role of proinflammatory cytokine interleukin-33 (IL-33) in the development of atopic diseases. Although, little is known about the role of IL-33 in virus-induced BA exacerbations.. We used mouse models of RSV (respiratory syncytial virus)-induced inflammation exacerbation in OVA-sensitized mice and RSV infection alone in adult animals to characterize expression of il33 in the mouse lungs. Moreover, we studied the influence of il33 knockdown with intranasally administrated siRNA on the development of RSV-induced inflammation exacerbation. In addition, we evaluated the expression of IL33 in the ex vivo stimulated PBMCs from allergic asthma patients and healthy subjects with and without confirmed acute respiratory viral infection.. Using mouse models, we found that infection with RSV drives enhanced il33 mRNA expression in the mouse lung. Treatment with anti-il33 siRNA diminishes airway inflammation in the lungs (we found a decrease in the number of inflammatory cells in the lungs and in the severity of histopathological alterations) of mice with RSV-induced inflammation exacerbation, but do not influence viral load. Elevated level of the IL33 mRNA was detected in ex vivo stimulated blood lymphocytes of allergic asthmatics infected with respiratory viruses. RSV and rhinovirus were the most detected viruses in volunteers with symptoms of respiratory infection.. The present study provides additional evidence of the crucial role of the IL-33 in pathogenesis of RSV infection and virus-induced allergic bronchial asthma exacerbations.

Topics: Adolescent; Adult; Aged; Animals; Asthma; Disease Models, Animal; Female; Humans; Hypersensitivity; Inflammation; Interleukin-33; Leukocytes, Mononuclear; Lung; Male; Mice; Mice, Inbred BALB C; Middle Aged; Ovalbumin; Respiratory Syncytial Virus Infections; Respiratory Syncytial Virus, Human; Respiratory Syncytial Viruses; RNA, Messenger; RNA, Small Interfering; Up-Regulation; Young Adult

Cysteinyl leukotrienes (CysLTs), a group of inflammatory lipid mediators, are found elevated in obese-asthmatic patients. Leukotriene D. Primary human small airway epithelial cells (SAECs) were stimulated with different concentrations of LTD

Topics: Airway Remodeling; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cells, Cultured; Cytokines; Epithelial Cells; Humans; Inflammasomes; Inflammation; Leukocyte Count; Leukotriene D4; Male; Mice, Inbred BALB C; Mucin 5AC; NLR Family, Pyrin Domain-Containing 3 Protein; Obesity; Ovalbumin; Smad2 Protein; Smad3 Protein; Vimentin

Asthma is an allergic chronic inflammatory disease of the pulmonary airways, characterized by the infiltration of white blood cells and release of inflammatory cytokines of complex pathways linked to its pathogenesis. Syringin extracted from various medicinal plants has been used extensively for the treatment of inflammatory diseases. Hence, this study was conducted to further explore the protective effects of the syringin in ovalbumin (OVA) induced-asthma mice model. OVA-sensitized BALB mice were treated intraperitonealy with three doses (25, 50 and 100 mg/kg) of the syringin which was validated by the alteration in the immunoglobulin E (IgE) levels, cytokines levels, histopathological evaluation inflammatory cell count, lung weight, nitrite (NO) levels, oxidative stress biomarkers and gene markers. The treatment of syringin intensely reduced the increased IgE, inflammatory cytokines, WBC count and restored the antioxidant stress markers OVA stimulated animals. In addition, a significant reduction in inflammation and mucus production was evidenced in histopathological analysis which was further validated by suppression NF-κB pathway activation by syringin. These results suggest that syringin may improve asthma symptoms in OVA-induced mice by modulating NF-κB pathway activation.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Disease Models, Animal; Female; Glucosides; Inflammation; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Phenylpropionates; Pneumonia; Signal Transduction

The use of probiotics has been broadly popularized due to positive effects in the attenuation of aberrant immune responses such as asthma. Allergic asthma is a chronic respiratory disease characterized by airway inflammation and remodelling.. This study was aimed to evaluate the effect of oral administration of Lactococcus lactis NZ9000 on asthmatic airway inflammation and lung tissue remodelling in rats and its relation to the maintenance of an adequate intestinal barrier.. Wistar rats were ovalbumin (OVA) sensitized and challenged and orally treated with L. lactis. Lung inflammatory infiltrates and cytokines were measured, and remodelling was evaluated. Serum OVA-specific immunoglobulin (Ig) E levels were assessed. We also evaluated changes on intestinal environment and on systemic immune response.. L. lactis diminished the infiltration of proinflammatory leucocytes, mainly eosinophils, in the bronchoalveolar compartment, decreased lung IL-4 and IL-5 expression, and reduced the level of serum allergen-specific IgE. Furthermore, L. lactis prevented eosinophil influx, collagen deposition, and goblet cell hyperplasia in lung tissue. In the intestine, L. lactis-treated asthmatic rats increased Peyer's patch and goblet cell quantity and mRNA expression of IgA, MUC-2, and claudin. Additionally, intestinal morphological alterations were normalized by L. lactis administration. Splenocyte proliferative response to OVA was abolished, and serum levels of transforming growth factor (TGF)-β were increased by L. lactis treatment.. These findings suggest that L. lactis is a potential candidate for asthma prevention, and the effect is mediated by the improvement of intestinal barrier function and systemic TGF-β production.

Topics: Airway Remodeling; Animals; Asthma; Cytokines; Disease Models, Animal; Immunoglobulin E; Inflammation Mediators; Intestinal Mucosa; Lactococcus lactis; Leukocytes; Ovalbumin; Probiotics; Rats; Transforming Growth Factor beta

Allergic asthma is a chronic inflammatory disease of the lung and the airway, which is characterized by aberrant type 2 immune responses to otherwise unharmful aeroallergens. While the central role of Th2 cells and type 2 cytokines in the pathogenesis of allergic asthma is well documented, the regulation and plasticity of Th2 cells remain incompletely understood. By using an animal model of allergic asthma in IL-4-reporter mice, we found that Th2 cells in the lung expressed higher levels of Rora than those in the lymph nodes, and that treatment with an RORα agonist SR1078 resulted in diminished Th2 cell responses in vivo. To determine the T cell-intrinsic role of RORα in allergic asthma in vivo, we established T cell-specific RORα-deficient (Cd4creRora

Topics: Administration, Intranasal; Animals; Aspergillus oryzae; Asthma; Benzamides; CD8-Positive T-Lymphocytes; Cell Differentiation; Disease Models, Animal; Interleukin-4; Lung; Mice; Nuclear Receptor Subfamily 1, Group F, Member 1; Ovalbumin; Th2 Cells

Topics: Animals; Asthma; Disease Models, Animal; Embryonic Development; Estrogens; Female; Humans; Mice; Ovalbumin; Oxidative Stress; Pregnancy; Pregnancy Complications; Progesterone

Steroids are the main drugs currently used to treat asthma. However, the toxic and side effects of these drugs and the tolerance of the drugs due to long-term administration are still problems in the clinical treatment of asthma. Bavachinin has a good effect in the treatment of mouse asthma models, and it can effectively inhibit the expression of a variety of cytokines. However, it is extremely difficult to dissolve in water, has low bioavailability, and is quickly cleared in the blood. These characteristics limit its clinical application potential. In this study, nanotechnology was used to construct an effective oral drug delivery system. Through analysis of serum-related antibodies and cytokines, the system showed significant therapeutic effects on asthma-positive groups. Far-infrared imaging results showed that the system has a good targeted enrichment effect on pathological parts, while showing lower toxicity and higher therapeutic effect. Whether it is the splenocyte flow typing or the analysis of lung tissue, the system has verified the excellent treatment, and through the observation of paraffin sections of lung tissue, it was found that the bronchial morphology returned to normal after drug treatment, and the leakage of inflammatory cells was significantly reduced.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Flavonoids; Lung; Mice; Mice, Inbred BALB C; Nanospheres; Ovalbumin

Mouse models of allergic asthma have been utilized to establish the role of T helper type 2 (Th2) cells in driving lung inflammation, airway hyperresponsiveness, and obstruction. Here, we present the allergic asthma models, in which mice are hypersensitized to ovalbumin (OVA) and house dust mite (HDM). These models mimic the major characteristics of human asthma including the eosinophilic inflammation and hyperactivity of the airway, overproduction of Th2 cytokines in the lung, and elevated total and allergen-specific immunoglobulin E (IgE) in serum.

Topics: Adjuvants, Immunologic; Allergens; Aluminum Hydroxide; Animals; Asthma; Biomarkers; Dendritic Cells; Disease Models, Animal; Eosinophils; Flow Cytometry; Forkhead Transcription Factors; Gene Expression; Immunoglobulin E; Interferon-gamma; Interleukins; Lung; Macrophages; Mice; Mice, Inbred C57BL; Ovalbumin; Pyroglyphidae; Respiratory Function Tests; Respiratory Hypersensitivity; Th2 Cells

Polymorphonuclear neutrophils (PMN) are one fraction of the major inflammatory cells in allergic asthma (asthma, in short); the role of PMN in the asthma pathogenesis is not fully understood yet. This study aims to investigate the effects of specific Ag-guiding exosomes on suppressing the neutrophil-dominant airway inflammation. In this study, BALB/c mice were immunized with ovalbumin plus complete Freund adjuvant to induce an asthma model featured with neutrophil-dominant lung inflammation. The Ag specific PMN (sPMN)-targeting exosomes (tExo), that were exosomes carrying a complex of specific Ag/anti-CD64 Ab and Fas ligand, were constructed to be used to alleviate neutrophilic asthma in mice. We found that sPMNs were the major cellular component in bronchoalveolar lavage fluid (BALF) in asthma mice, while less than 3% PMNs in naive control mice. The sPMNs expressed higher levels of CD64, which formed complexes with Ag-specific IgG (sIgG). The sIgG/CD64 complex-carrying PMNs could be activated upon exposing to specific Ags. Exposure to tExos induced Ag-specific PMNs apoptosis. Administration of tExos efficiently suppressed experimental asthma. We conclude that a fraction of sPMN was identified in the airway of asthma mice. The sPMNs could be activated upon exposure to specific Ags. tExos could induce sPMNs apoptosis, that show the translational potential in the treatment of asthma.

Topics: Animals; Antibodies; Antigens; Apoptosis; Asthma; Bronchoalveolar Lavage Fluid; Drug Carriers; Exosomes; Freund's Adjuvant; Hypersensitivity; Immunoglobulin G; Lung; Mice; Mice, Inbred BALB C; Nanoparticles; Neutrophils; Ovalbumin; Pneumonia; Receptors, IgG; Vaccines

Asthma is a chronic inflammatory lung disorder with continuously increasing prevalence worldwide. Novel strategies are needed to prevent or improve asthma. The aim of this study was to investigate the effects of sophoricoside from Sophora japonica on allergic asthma. The mature seeds of S. japonica contain a large amount of sophoricoside. Sophoricoside reduced allergic and asthmatic symptoms by suppressing airway inflammation and antibody-antigen reaction in mouse models. In particular, sophoricoside suppressed immune cell recruitment into the airway lumens of the lungs and production of pro-inflammatory cytokines in the bronchoalveolar lavage fluid (BALF) of ovalbumin (OVA)-induced mice. It also decreased the amounts of histamine and arachidonic acid metabolites released in OVA-induced mice and antibody-antigen stimulated mast cells. In addition, sophoricoside decreased differentiation of naïve CD4

Topics: Animals; Anti-Allergic Agents; Anti-Asthmatic Agents; Asthma; Benzopyrans; CD4-Positive T-Lymphocytes; Cell Degranulation; Cell Differentiation; Cells, Cultured; Cytokines; Disease Models, Animal; Histamine Release; Immunoglobulins; Inflammation Mediators; Lung; Mast Cells; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Sophora

Asthma is a chronic airway inflammatory disease caused by a variety of cytokines and signaling pathways closely related to immunoregulation. Corticosteroids are the most widely used drug in the asthma treatment. However, the use of corticosteroids could cause topical side effects. So, it's important to find new drugs for asthma treatment. Our study aims to explore the pharmacological effect of borneol on asthma and its underlying mechanism.. We constructed the OVA-induced asthma model to investigate the effect of borneol on asthma in mice. HE and PAS staining was used to detect the effect of borneol on pathological change of mice with asthma. Inflammatory cytokines were measured by ELISA. qRT-PCR was used to explore the effect of borneol on microRNAs expression. Cell proliferation of CD4 + T cells was detected by CCK-8 assay and flow cytometry. Western blot was used to detect pten expression and Akt activation.. We found that borneol significantly alleviated asthma progression in mice. Borneol inhibited CD4 + T cells infiltration in vivo and proliferation in vitro by downregulating miR-26a and miR-142-3p. miR-26a and miR-142-3p promoted CD4 + T cells proliferation in vitro through targeting Pten. Overexpression of miR-26a and miR-142-3p abolished the effect of borneol in vivo.. In a word, these findings suggested that borneol attenuated asthma in mice by decreasing the CD4 + T cells infiltration. The molecular mechanism of borneol was dependent on the downregulation of miR-26a and miR-142-3p to upregulate the Pten expression.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Camphanes; CD4-Positive T-Lymphocytes; Cell Proliferation; Disease Models, Animal; Down-Regulation; Lung; Male; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; PTEN Phosphohydrolase; Signal Transduction

Growing evidence indicates insufficient autophagy is crucial to airway remodeling in asthma. However, it is uncertain whether p62, an autophagy major regulator, mediates the airway remodeling process. This study aimed to evaluate the role and underlying mechanism of p62 in airway remodeling in asthma.. Airway remodeling was confirmed via histopathology. Western blotting and RT-PCR were used to detect the expression of autophagic and glycolytic proteins, as well as glycolytic genes. Glycolysis was measured by glucose consumption and lactate production. Cell proliferation was analyzed by CCK8 assays while and the scratch test and transwell method were used for cell migration.. We found that insufficient autophagic flux and increased p62 expression existed in chronic asthma mice. Additionally, knockdown of p62 inhibited asthmatic human bronchial smooth muscle cells (BSMCs) proliferation and migration in vitro. To elucidate the underlying mechanism of p62-mediated autophagy flux in directing BSMCs function, we demonstrated that knockdown of p62 decreased the glucose consumption and lactate production in BSMCs, whereas p62 overexpression had the opposite effect. Furthermore, we showed that p62 regulated glycolysis in BSMCs by the mTOR/c-Myc/hexokinase 2 (HK2) pathway.. Our findings suggest that p62 is involved in BSMCs proliferation and migration via the mTOR/c-Myc/HK2-mediated glycolysis, thereby providing a new target for airway remodeling treatment.

Topics: Airway Remodeling; Animals; Asthma; Autophagy; Cell Movement; Cell Proliferation; Cellular Reprogramming; Disease Models, Animal; Female; Glycolysis; Hexokinase; Mice; Mice, Inbred BALB C; Myocytes, Smooth Muscle; Ovalbumin; Proto-Oncogene Proteins c-myc; Sequestosome-1 Protein; TOR Serine-Threonine Kinases

Asthma is a chronic airway inflammatory disease and acupuncture is frequently used in patients suffering from asthma in clinic. However, the regulatory mechanism of acupuncture treatment in asthma is not fully elucidated. We sought to investigate the effectiveness of acupuncture on asthma and the associated regulatory mechanism. An ovalbumin (OVA)-induced mouse asthma model was established and the effect of acupuncture on airway hyperresponsiveness (AHR), mucus hypersecretion and inflammation was assessed. Tandem mass tag (TMT)-based quantitative proteomics analysis of lung tissue and bioinformatics analysis were performed. Our results revealed that the OVA-induced mouse asthma model was successfully established with the significantly elevated AHR to methacholine (Mch), and acupuncture was effective in attenuation of AHR to Mch, peribronchial and perivascular inflammation and mucus production. The inflammatory cells around the airways, mucous secretion as well as levels of IgE, CCL5, CCL11, IL-17A in bronchoalveolar lavage fluid (BALF) and IL-4, IL-5 and IL-13 levels in serum were siginificantly inhibited by acupuncture. TMT-based quantitative proteomics analysis found that a total of 6078 quantifiable proteins were identified, and 564 (334 up-regulated and 230 down regulated) differentially expressed proteins (DEPs) were identified in OVA-induced asthma model group (A) versus normal control group (NC). Acupuncture treatment resulted in 667 DEPs (416 up-regulated and 251 down regulated) compared with A group, and 86 overlapping DEPs were identified in NC, A and AA groups. Among the 86 overlapping DEPs, we identified 41 DEPs regulated by acupuncture. Based on the above data, we performed a systematic bioinformatics analysis of the 41 DEPs, and results showed that these 41 DEPs were predominantly related to 4 KEGG pathways including SNARE interactions in vesicular transport, ferroptosis, endocrine and other factor-regulated calcium reabsorption, and protein digestion and absorption. DEPs of SLC3A2 and ATP1A3 expression levels were verified by immumohistochemical staining. Mice in OVA-induced asthma model group had elevated SLC3A2 and ATP1A3 expression and acupuncture had the ability to downregulate SLC3A2 and ATP1A3 protein expression. Furthermore, acupuncture reduced the MDA level and increased the GSH and SOD levels in the lung tissue. Taken together, our data suggested that acupuncture was effective in treating asthma by attenuation of AHR, mucus secretion

Topics: Acupuncture Therapy; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Fusion Regulatory Protein 1, Heavy Chain; Inflammation; Lung; Methacholine Chloride; Mice; Mice, Inbred BALB C; Mucus; Ovalbumin; Proteomics; Respiratory Hypersensitivity; Sodium-Potassium-Exchanging ATPase

Evidence proved that gestational ozone (O. This study aimed to investigate whether gestational O. The pregnant ICR mice were randomly grouped and were administered O. OVA sensitization and challenge successfully induced asthma in offspring. Compared with the air control, pulmonary inflammation infiltration, mucous secretion, the concentration of OVA-specific IgE, the level of tumor necrosis factor (TNF)-α, and T helper (Th) 2-skewed response were significantly exacerbated when O. Exposure to O

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred ICR; Ovalbumin; Pregnancy

Growing evidence shows that enhancer of zeste homolog 2 (EZH2) plays a role in various physiological functions and cancer pathogenesis. However, its contribution to allergic diseases remains controversial. We sought to investigate the role of EZH2 in the pathogenesis of allergic airway inflammation.. 3-Deazaneplanocin A (DZNep), an indirect inhibitor of EZH2, was administered via intraperitoneal injection in an ovalbumin (OVA)-induced murine model of allergic airway inflammation. The expression of EZH2 in the allergic airway tissues was examined by immunohistochemistry (IHC) and western blot. The inflammatory cell infiltration and the goblet cell hyperplasia in the murine nose and lung were detected by hematoxylin and eosin (H&E) staining and periodic acid-Schiff (PAS) staining. Levels of cytokines, including IL-4, IFN-γ, IL-6, and IL-10, were evaluated in the bronchoalveolar lavage fluid (BALF) using Enzyme-linked immune sorbent assay (ELISA).. EZH2 expression was inhibited by DZNep treatment (P < 0.05). The administration of DZNep significantly inhibited the inflammatory cell infiltration (P < 0.0001) and goblet cell hyperplasia (P < 0.001). Moreover, it suppressed the secretion of IL-4 (P < 0.0001) and IL-6 (P < 0.01) in the BALF.. Our findings demonstrate that DZNep attenuates allergic airway inflammation and could be a new therapeutic option for allergic rhinitis and asthma.

Topics: Adenosine; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Hanchuan Zupa Granule (HCZP), a traditional Chinese ethnodrug, has the functions of supressing a cough, resolving phlegm, warming the lungs, and relieving asthma. In clinical practice employing traditional Chinese medicine (TCM), HCZP is commonly used to treat acute colds, cough and abnormal mucous asthma caused by a cold, or "Nai-Zi-Lai" in the Uygur language. Studies have confirmed the use of HCZP to treat cough variant asthma (CVA) and other respiratory diseases. However, the pharmacological mechanisms of HCZP remain unrevealed.. To investigate the anti-tussive and anti-asthmatic effects and the possible pharmacological mechanisms of HCZP in the treatment of CVA.. A guinea pig CVA animal model was established by intraperitoneal injection of ovalbumin (OVA) combined with intraperitoneal injection of aluminium hydroxide adjuvant and atomized OVA. Meanwhile, guinea pigs with CVA received oral HCZP (at dosages of 0.571, 0.285 and 0.143 g/kg bodyweight). The number of coughs induced by aerosol capsaicin was recorded, and the airway hyperresponsiveness (AHR) of CVA guinea pigs was detected with the FinePointe series RC system. H&E staining of lung tissues was performed to observe pathological changes. ELISA was used to detect inflammatory cytokines. qRT-PCR and western blotting analyses were used to detect the expression of Th1-specific transcription factor (T-bet), Th2-specific transcription factor (GATA3), and Toll-like receptor 4 (TLR4) signal transduction elements. These methods were performed to assess the protective effects and the potential mechanisms of HCZP on CVA.. Great changes were found in the CVA guinea pig model after HCZP treatment. The number of coughs induced by capsaicin in guinea pigs decreased, the body weights of guinea pigs increased, and inflammation of the eosinophilic airway and AHR were reduced simultaneously. These results indicate that HCZP has a significant protective effect on CVA. A pharmacological study of HCZP showed that the levels of interleukin-4 (IL-4) and IL-5 and tumour necrosis factor-α (TNF-α) in serum decreased. The amount of interferon-γ (IFN-γ) increased, mRNA and protein expression of TLR4 and GATA3 weakened, and mRNA and protein expression of T-bet increased.. HCZP ameliorated the symptoms of guinea pigs with CVA induced by OVA by regulating the Th1/Th2 imbalance and TLR4 receptors.

Topics: Animals; Anti-Asthmatic Agents; Antitussive Agents; Asthma; Body Weight; Capsaicin; Cough; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Flavonoids; GATA3 Transcription Factor; Glycyrrhizic Acid; Guinea Pigs; Lung; Medicine, Chinese Traditional; Ovalbumin; Respiratory Hypersensitivity; T-Box Domain Proteins; Th1 Cells; Th2 Cells; Toll-Like Receptor 4; Triterpenes

The airway epithelium is continuously exposed to a variety of pollutants and allergens, thanks to both natural and manmade environmental pollution. With numerous protective mechanisms, the airway epithelium protects the lungs. DNA repair mechanism is one such protective response and its failure could lead to the accumulation of DNA mutations. Our lab had earlier demonstrated the dysfunctional mitochondria in airway epithelium of the asthmatic mice lungs. Here, we show that Ku70 modulation by the administration of Ku70 plasmid attenuates asthma features and reduces mitochondrial dysfunction in the lungs of allergen exposed mice. Ku70 is a key DNA repair protein with diverse roles including VDJ recombination, telomere maintenance, and maintenance of cell homeostasis. Recently, we found a reduction in Ku70 expression in asthmatic airway epithelium, and this was associated with mitochondrial dysfunction in asthmatic condition. In this study, we have shown that Ku70 over-expression in asthmatic mice attenuated airway hyperresponsiveness, airway inflammation, sub-epithelial fibrosis along with reduction in TGF-β with no effect in IL-13 levels and goblet cell metaplasia. Ku70 over-expression in asthmatic mice reduced 8-isoprostane, a marker of oxidative stress, and restored the mitochondrial function in asthmatic mice. We further found these roles of Ku70 to be independent of DNA damage as Ku70 overexpressed mice did not show any reduction in DNA tail, an index of DNA damage. Thus, our findings indicate that Ku70 can attenuate crucial features of asthma along with the restoration of mitochondrial function. This implies that Ku70 could be a therapeutic target for asthma without affecting DNA repair function.

Topics: Animals; Asthma; Bronchoalveolar Lavage; Disease Models, Animal; Genetic Vectors; Injections, Intravenous; Ku Autoantigen; Lung; Male; Mice; Mitochondria; Ovalbumin; Plasmids; Transforming Growth Factor beta

Traditionally, the roots of Angelica reflexa B.Y.Lee (AR) have been used to treat cough, phlegm, neuralgia, and arthralgia in Northeast Asia.. The anti-asthmatic effect of AR root extract (ARE) was determined using a murine airway allergic inflammation model and the primary T cell polarization assay.. To evaluate the anti-asthmatic effect of ARE, inflammatory cell infiltration was determined histologically and inflammatory mediators were measured in bronchoalveolar lavage fluid (BALF). Furthermore, the effects of AREs on Th2 cell differentiation and activation were determined by western blotting and flow cytometry.. Asthmatic phenotypes were alleviated by ARE treatment, which reduced mucus production, inflammatory cell infiltration (especially eosinophilia), and type 2 cytokine levels in BALF. ARE administration to mice reduced the number of activated Th2 (CD4. Our findings indicate that the anti-asthmatic effect of AREs is mediated by the reduction in Th2 cell activation by regulating IRF4.

Topics: Angelica; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; GATA3 Transcription Factor; Hypersensitivity; Interferon Regulatory Factors; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Ovalbumin; Plant Extracts; Plant Roots; Pneumonia; Pulmonary Eosinophilia; RAW 264.7 Cells; Th2 Cells

Mesenchymal stem cells (MSCs) have been investigated in preventing and treating allergic asthma in many reports. Recently, MSC-derived exosomes (MSC-Exo) were showed a promising alternative to stem cell-based therapy in many kinds of diseases. However, the effect of MSC-Exo on allergic asthma has not been investigated thoroughly thus far. Here, we aimed to investigate the immunomodulation effect of MSC-Exo in a murine model of asthma and explore the underlying mechanisms. BALB/c mice were sensitized and challenged by OVA to establish asthma model. MSC-Exo were intranasally delivered before or during challenge and the protective effect were assessed after the last OVA challenge. Allergic airway inflammation elicited by OVA were significantly attenuated by intranasal delivery of MSC-Exo. To explore the protective mechanism of MSC-Exo, lung interstitial macrophages (IMs) and alveolar macrophages (AMs) were analyzed by flow cytometry and the origin of IMs were traced. Lung IMs ratios were significantly enhanced and high level of IL-10 was produced after MSC-Exo intranasal delivery. IMs ratios were not obviously affected by CCR2 inhibitor or Clodronate liposome administration, whereas significantly decreased in splenectomized mice. Cx3cr1

Topics: Animals; Asthma; Cell Proliferation; Cells, Cultured; CX3C Chemokine Receptor 1; Disease Models, Animal; Exosomes; Interleukin-10; Lung; Macrophages, Alveolar; Mesenchymal Stem Cell Transplantation; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Spleen; Splenectomy

Asthma is characterized by airway hyperresponsiveness (AHR), inflammation, and airway remodeling. Airway hyperresponsiveness results from enhanced airway smooth muscle (ASM) contraction potentially under the control of an epithelium-derived relaxing factor (EpDRF). However, relatively rare is known about EpDRF. We aimed to elucidate the role of epithelium-derived stanniocalcin-1 (STC1) on AHR and ASM contraction.. Stanniocalcin-1 levels in the serum of asthmatic patients and healthy volunteers and in bronchoalveolar lavage fluid (BALF) from ovalbumin (OVA)-challenged mice were measured by ELISA. The effects of exogenous STC1 on AHR and on inflammation were examined in mice. IL-13 modulation of STC1 mRNA and protein levels was studied in human bronchial epithelial cell lines (16HBE). The function of STC1 on Ca. Serum STC1 was decreased in asthma (n = 93) compared with healthy volunteers (1071 ± 30.4 vs 1414 ± 75.1 pg/ml, p < 0.0001, n = 23) and correlated with asthma control (p = 0.0270), lung function (FEV1, p = 0.0130), and serum IL-13 levels (p = 0.0009). Treatment of ten asthmatic subjects with inhaled corticosteroids/long-acting beta2-agonists (ICS/LABA) for 1 year enhanced STC1 expression which correlated with improved asthma control (p = 0.022). STC1 was mainly expressed in bronchial epithelium and intranasal administration of recombinant human STC1 (rhSTC1) reduced AHR and inflammation in mice. IL-13 suppressed STC1 release from 16HBE, whereas rhSTC1 blocked store-operated Ca. Our data indicate that STC1 deficiency in asthmatic airways promotes STIM1 hyperactivity, enhanced ASM contraction, and AHR. STC1 may be a candidate EpDRF.

Topics: Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Calcium Channels; Glycoproteins; Humans; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity

As one of the main functional forms of mesenchymal stem cells (MSCs), MSC-derived extracellular vesicles (MSC-EVs) have shown an alternative therapeutic option in experimental models of allergic asthma. Oxygen concentration plays an important role in the self-renewal, proliferation, and EV release of MSCs and a recent study found that the anti-asthma effect of MSCs was enhanced by culture in hypoxic conditions. However, the potential of hypoxic MSC-derived EVs (Hypo-EVs) in asthma is still unknown.. BALB/c female mice were sensitized and challenged with ovalbumin (OVA), and each group received PBS, normoxic human umbilical cord MSC-EVs (Nor-EVs), or Hypo-EVs weekly. After treatment, the animals were euthanized, and their lungs and bronchoalveolar lavage fluid (BALF) were collected. With the use of hematoxylin and eosin (HE), periodic acid-Schiff (PAS) and Masson's trichrome staining, enzyme-linked immune sorbent assay (ELISA), Western blot analysis, and real-time PCR, the inflammation and collagen fiber content of airways and lung parenchyma were investigated.. Hypoxic environment can promote human umbilical cord MSCs (hUCMSCs) to release more EVs. In OVA animals, the administration of Nor-EVs or Hypo-EVs significantly ameliorated the BALF total cells, eosinophils, and pro-inflammatory mediators (IL-4 and IL-13) in asthmatic mice. Moreover, Hypo-EVs were generally more potent than Nor-EVs in suppressing airway inflammation in asthmatic mice. Compared with Nor-EVs, Hypo-EVs further prevented mouse chronic allergic airway remodeling, concomitant with the decreased expression of pro-fibrogenic markers α-smooth muscle actin (α-SMA), collagen-1, and TGF-β1-p-smad2/3 signaling pathway. In vitro, Hypo-EVs decreased the expression of p-smad2/3, α-SMA, and collagen-1 in HLF-1 cells (human lung fibroblasts) stimulated by TGF-β1. In addition, we showed that miR-146a-5p was enriched in Hypo-EVs compared with that in Nor-EVs, and Hypo-EV administration unregulated the miR-146a-5p expression both in asthma mice lung tissues and in TGF-β1-treated HLF-1. More importantly, decreased miR-146a-5p expression in Hypo-EVs impaired Hypo-EV-mediated lung protection in OVA mice.. Our findings provided the first evidence that hypoxic hUCMSC-derived EVs attenuated allergic airway inflammation and airway remodeling in chronic asthma mice, potentially creating new avenues for the treatment of asthma.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Extracellular Vesicles; Female; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Obesity increases the morbidity and severity of asthma, with poor sensitivity to corticosteroid treatment. Metformin has potential effects on improving asthma airway inflammation. Regulatory T cells (Tregs) play a key role in suppressing the immunoreaction to allergens. We built an obese asthmatic mouse model by administering a high-fat diet (HFD) and ovalbumin (OVA) sensitization, with daily metformin treatment. We measured the body weight and airway inflammatory status by histological analysis, qRT-PCR, and ELISA. The percentage of Tregs was measured by flow cytometry. Obese asthmatic mice displayed more severe airway inflammation and more significant changes in inflammatory cytokines. Metformin reversed the obese situation and alleviated the airway inflammation and remodelling with increased Tregs and related transcript factors. The anti-inflammatory function of metformin may be mediated by increasing Tregs.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Body Weight; Bronchoalveolar Lavage Fluid; CD4 Lymphocyte Count; Diet, High-Fat; Disease Models, Animal; Humans; Hypoglycemic Agents; Inflammation; Interleukin-4; Lung; Metformin; Mice; Obesity; Ovalbumin; Spleen; T-Lymphocytes, Regulatory; Tumor Necrosis Factor-alpha

The objective of this study was to investigate the inhibitory effect of miR-135a in regulating JAK/STAT signaling pathway on airway inflammation in asthmatic mice. An asthma model was established by sensitization and stimulation with ovalbumin (OVA), and the corresponding drug intervention was given from the day of stimulation by means of nasal drops. Airway hyperresponsiveness was tested. The content of miR-135a in the lung tissue of mice was detected by RT-PCR. The pathological changes of lung tissue were evaluated by HE staining. Tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-5, and eotaxin in bronchoalveolar lavage fluid (BALF) and lung tissue were detected by ELISA and immunohistochemistry, respectively. The expression of JAK/STAT signaling pathway-related protein in lung tissue was detected by western blot. To further validate the effect of miR-135a overexpression on the JAK/STAT signaling pathway, pathway activators and inhibitors were added. Compared with the OVA group, the airway hyperresponsiveness of the mice was significantly decreased after treatment with the miR-135a agonist. The expression of miR-135a was significantly increased in the lung tissue and the pathological changes of the lung tissue were alleviated. The contents of TNF-α, IL-6, IL-5, and eotaxin in BALF and lung tissues were decreased. The expression of JAK/STAT signaling pathway-related proteins p-JAK3/JAK3, p-STAT1/STAT1, and p-STAT3/STAT3 were significantly reduced in lung tissue (P<0.05). Addition of JAK inhibitor AG490 reduced airway inflammation in asthmatic mice. miR-135a agonists inhibit airway inflammation in asthmatic mice by regulating the JAK/STAT signaling pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Lung; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Signal Transduction

Substantial recent advances in the comprehension of the molecular and cellular mechanisms behind asthma have evidenced the importance of the lung immune environment for disease outcome, making modulation of local immune responses an attractive therapeutic target against this pathology. Live attenuated mycobacteria, such as the tuberculosis vaccine BCG, have been classically linked with a type 1 response, and proposed as possible modulators of the type 2 response usually associated with asthma.. In this study we used different acute and chronic murine models of asthma to investigate the therapeutic efficacy of intranasal delivery of the live tuberculosis vaccines BCG and MTBVAC by regulating the lung immune environment associated with airway hyperresponsiveness (AHR).. Intranasal administration of BCG, or the novel tuberculosis vaccine candidate MTBVAC, abrogated AHR-associated hallmarks, including eosinophilia and lung remodeling. This correlated with the re-polarization of allergen-induced M2 macrophages towards an M1 phenotype, as well as with the induction of a strong allergen-specific Th1 response. Importantly, vaccine treatment was effective in a scenario of established chronic asthma where a strong eosinophil infiltration was already present prior to immunization. We finally compared the nebulization efficiency of clinical formulations of MTBVAC and BCG using a standard commercial nebulizer for potential aerosol application.. Our results demonstrate that pulmonary live tuberculosis vaccines efficiently revert established asthma in mice. These data support the further exploration of this approach as potential therapy against asthma.. Spanish Ministry of Science [grant numbers: BIO2014-5258P, RTI2018-097625-B-I00], Instituto de Salud Carlos III, Gobierno de Aragón/Fondo Social Europeo, University of Zaragoza [grant number: JIUZ-2018-BIO-01].

Topics: Administration, Intranasal; Airway Remodeling; Allergens; Animals; Asthma; BCG Vaccine; Biomarkers; Cellular Microenvironment; Cytokines; Disease Models, Animal; Eosinophils; Female; Immunization; Mice; Ovalbumin; Tuberculosis Vaccines; Vaccines, Attenuated

There is growing inclination towards developing bioactive molecule-based strategies for the management of allergic airway inflammation associated respiratory diseases. Vitex negundo Linn., also known as Nirgundi, is one such medicinal plant enriched with phytochemicals and used for inflammatory and respiratory disorders including asthma in traditional system of medicine. Preliminary studies have claimed anti-tussive and bronchodilator potential of V. negundo Linn. However, its attributes as well as molecular mechanism (s) in modulation of asthma mediated by allergic inflammation are yet to be delineated scientifically.. Present study attempted to assess the effectiveness of Vitex negundo leaf extract (VNLE) in mitigation of allergen induced inflammation associated asthmatic lung damage with emphasis to delineate its molecular mechanism (s).. Allergic lung inflammation was established in Balb/c mice using Ovalbumin-lipopolysaccharide (OVA-LPS). Several allergic inflammatory parameters, histopathological changes, alveolar macrophage activation and signalling pathways were assessed to examine protective effects of VNLE. UHPLC-DAD-QTOF-ESI-IMS was used to characterize VLNE.. VNLE administration effectively attenuated LPS-induced oxi-inflammatory stress in macrophages suggesting its anti-inflammatory potential. Further, VNLE showed protective effect in mitigating asthmatic lung damage as evident by reversal of pathological changes including inflammatory cell influx, congestion, fibrosis, bronchial thickness and alveolar collapse observed in allergen group. VNLE suppressed expressions of inflammatory Th1/Th2 cytokines, chemokines, endopeptidases (MMPs), oxidative effector enzyme (iNOS), adhesion molecules, IL-4/IFN-γ release with simultaneous enhancement in levels of IL-10, IFN-γ, MUC3 and tight junction proteins. Subsequent mechanistic investigation revealed that OVA-LPS concomitantly enhanced phosphorylation of NF-κB, PI3K, Akt and p38MAPKs and downregulated AMPK which was categorically counteracted by VNLE treatment. VNLE also suppressed OVA-LPS induced fibrosis, apoptosis, autophagy and gap junction proteins which were affirmed by reduction in TGF-β, Smad2/3/4, Caspase9/3, Bax, LC3A/B, connexin 50, connexin 43 and enhancement in Bcl2 expression. Additionally, suppression of alveolar macrophage activation, inflammatory cells in blood and elevation of splenic CD8+T cells was demonstrated. UHPLC-DAD-QTOF-ESI-IMS revealed presence of iridoids glycoside and phenolics which might contribute these findings.. These findings confer protective effect of VNLE in attenuation of allergic lung inflammation and suggest that it could be considered as valuable medicinal source for developing safe natural therapeutics for mitigation of allergic inflammation during asthma.

Topics: AMP-Activated Protein Kinases; Animals; Anti-Inflammatory Agents; Asthma; Caspases; Disease Models, Animal; Inflammation; Lipopolysaccharides; Lung Injury; Macrophage Activation; Mice, Inbred BALB C; Microtubule-Associated Proteins; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Phosphatidylinositol 3-Kinases; Plant Extracts; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Signal Transduction; Smad Proteins; Transforming Growth Factor beta; Vitex

Asthma is a chronic respiratory disease in which the nervous system plays a central role. Sensory nerve activation, amongst others via Transient Receptor Potential Ankyrin 1 (TRPA1) channels, contributes to asthma characteristics including cough, bronchoconstriction, mucus secretion, airway hyperresponsiveness (AHR) and inflammation. In the current study, we evaluated the efficacy of the novel TRPA1 antagonist BI01305834 against AHR and inflammation in guinea-pig models of asthma.. First, a pilot study was performed in a guinea-pig model of allergic asthma to find the optimal dose of BI01305834. Next, the effect of BI01305834 on (1) AHR to inhaled histamine after the early and late asthmatic reaction (EAR and LAR), (2) magnitude of EAR and LAR and (3) airway inflammation was assessed. Precision-cut lung slices and trachea strips were used to investigate the bronchoprotective and bronchodilating-effect of BI01305834. Statistical evaluation of differences of in vivo data was performed using a Mann-Whitney U test or One-way nonparametric Kruskal-Wallis ANOVA, for ex vivo data One- or Two-way ANOVA was used, all with Dunnett's post-hoc test where appropriate.. A dose of 1 mg/kg BI01305834 was selected based on AHR and exposure data in blood samples from the pilot study. In the subsequent study, 1 mg/kg BI01305834 inhibited AHR after the EAR, and the development of EAR and LAR elicited by ovalbumin in ovalbumin-sensitized guinea pigs. BI01305834 did not inhibit allergen-induced total and differential cells in the lavage fluid and interleukin-13 gene expression in lung homogenates. Furthermore, BI01305834 was able to inhibit allergen and histamine-induced airway narrowing in guinea-pig lung slices, without affecting histamine release, and reverse allergen-induced bronchoconstriction in guinea-pig trachea strips.. TRPA1 inhibition protects against AHR and the EAR and LAR in vivo and allergen and histamine-induced airway narrowing ex vivo, and reverses allergen-induced bronchoconstriction independently of inflammation. This effect was partially dependent upon histamine, suggesting a neuronal and possible non-neuronal role for TRPA1 in allergen-induced bronchoconstriction.

Topics: Administration, Inhalation; Animals; Asthma; Bronchoconstriction; Bronchodilator Agents; Dose-Response Relationship, Drug; Guinea Pigs; Humans; Lung; Male; Organ Culture Techniques; Ovalbumin; Pilot Projects; TRPA1 Cation Channel

Obesity is a correctable factor for uncontrolled bronchial asthma. However, the effects of glucagon-like peptide-1 receptor (GLP-1R) agonist, a recently approved antiobestic drug, on airway hyperresponsiveness (AHR) and immune responses are not known.. Mice were fed with high-fat diet (HFD, 60% fat) for 8 weeks to induce obesity. Ovalbumin (OVA) sensitization and challenges were performed for 7 weeks. The mice were injected intraperitoneally with GLP-1R agonist 5 times a week for 4 weeks after OVA sensitization. After AHR measurement, expression of Th2, Th17 cytokines, and interleukin (IL)-33 were measured in BALF and lung tissues. Moreover, IL-1β and activity level of nucleotide oligomerization domain-like receptor protein 3 (NLRP3) were analyzed to investigate the mechanism of GLP-1R agonist on asthmatic airway inflammation.. HFD induced significant weight gain, OVA sensitization and challenge in obese mice made eosinophilic airway inflammation, and increased AHR. Treatment with GLP-1R agonist-induced weight loss suppressed eosinophilic airway inflammation and decreased AHR. Expression of IL-4, 5, and 33 was increased in BALF of obese asthma mice followed by a decrease in response to GLP-1R agonist treatment. Moreover, lung tissue H&E stain revealed that peribronchial inflammation induced by obesity and OVA was effectively suppressed by GLP-1R agonist. Expressions of NLRP3, activated caspase-1, and IL-1β were increased in lung tissues of obese asthma mice and demonstrated a decrease in response to GLP-1R agonist treatment.. GLP-1R agonist effectively induced weight loss, suppressed eosinophilic bronchial airway inflammation, and AHR in obese asthma mice. These effects were mediated by suppression of NLRP3 inflammasome activity and IL-1β. GLP-1R agonist is proposed as a novel anti-asthmatic agent targeting the obese asthmatics.

Topics: Animals; Asthma; Disease Models, Animal; Glucagon-Like Peptide 1; Inflammasomes; Inflammation; Mice; Mice, Inbred BALB C; Mice, Obese; NLR Family, Pyrin Domain-Containing 3 Protein; Obesity; Ovalbumin; Pharmaceutical Preparations

Defective absorption of acute allergic airway inflammation is involved in the initiation and development of chronic asthma. After allergen exposure, there is a rapid recruitment of macrophages around the airways, which promote acute inflammatory responses. The Ang-(1-7)/Mas receptor axis reportedly plays protective roles in various tissue inflammation and remodeling processes in vivo. However, the exact role of Mas receptor and their underlying mechanisms during the pathology of acute allergic airway inflammation remains unclear.. We investigated the role of Mas receptor in acute allergic asthma and explored its underlying mechanisms in vitro, aiming to find critical molecules and signal pathways.. Mas receptor expression was assessed in ovalbumin (OVA)-induced acute asthmatic murine model. Then we estimated the anti-inflammatory role of Mas receptor in vivo and explored expressions of several known inflammatory cytokines as well as phosphorylation levels of MAPK pathways. Mas receptor functions and underlying mechanisms were studied further in the human bronchial epithelial cell line (16HBE).. Mas receptor expression decreased in acute allergic airway inflammation. Multiplex immunofluorescence co-localized Mas receptor and EpCAM, indicated that Mas receptor may function in the bronchial epithelium. Activating Mas receptor through AVE0991 significantly alleviated macrophage infiltration in airway inflammation, accompanied with down-regulation of CCL2 and phosphorylation levels of MAPK pathways. Further studies in 16HBE showed that AVE0991 pre-treatment inhibited LPS-induced or anisomycin-induced CCL2 increase and THP-1 macrophages migration via JNK pathways.. Our findings suggested that Mas receptor activation significantly attenuated CCL2 dependent macrophage recruitments in acute allergic airway inflammation through JNK pathways, which indicated that Mas receptor, CCL2 and phospho-JNK could be potential targets against allergic airway inflammation.

Topics: Acute Disease; Angiotensin I; Animals; Asthma; Chemokine CCL2; Cytokines; Imidazoles; Inflammation; Macrophage Activation; Macrophages; Male; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Ovalbumin; Peptide Fragments; Phosphorylation; Proto-Oncogene Mas; Proto-Oncogene Proteins; Receptors, G-Protein-Coupled; Respiratory System

Allergic asthma is a common chronic inflammatory disease characterized by airway inflammation, mucus hypersecretion and airway remodeling. Autophagy is a highly conserved intracellular degradation pathway in eukaryotic cells. There is growing evidence suggesting that dysregulation of autophagy is involved in the pathological process of asthma. Luteolin is a typical flavonoid compound with anti-inflammatory, anti-allergic and immune-enhancing functions. Previous studies have shown that luteolin can attenuate airway inflammation and hypersensitivity in asthma. However, whether luteolin can play a role in treating asthma by regulating autophagy remains unclear. The aim of the present study was to evaluate the therapeutic effect of luteolin on ovalbumin (OVA)-induced asthmatic mice, observe its effect on the level of autophagy in lung tissues, and further elucidate its underlying mechanism. The results showed that OVA-induced mice developed airway hyperresponsiveness, mucus over-production and collagen deposition. The number of inflammatory cells, levels of interleukin (IL)-4, IL-5 and IL-13 in bronchoalveolar lavage fluid (BALF) and OVA-specific IgE in serum were significantly increased. Furthermore, the infiltration of inflammatory cells was observed along with the activation of autophagy in lung tissues. Luteolin treatment significantly inhibited the OVA-induced inflammatory responses and the level of autophagy in lung tissues as well. Moreover, luteolin activated the PI3K/Akt/mTOR pathway and inhibited the Beclin-1-PI3KC3 protein complex in lung tissues of asthmatic mice. In conclusion, this study explored the regulatory mechanism of luteolin on autophagy in allergic asthma, providing biologic evidence for its clinical application.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Asthma; Autophagy; Beclin-1; Bronchoalveolar Lavage Fluid; Cell Line; Cytokines; Female; Luteolin; Mice, Inbred BALB C; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Signal Transduction; TOR Serine-Threonine Kinases

Asthma is a chronic inflammatory disease that cannot be cured. Maresin 1 (MaR1) is a specific lipid synthesized by macrophages that exhibits powerful anti-inflammatory effects in various inflammatory diseases. The goal of this study was to evaluate the effect of MaR1 on allergic asthma using an ovalbumin- (OVA-) induced asthma model. Thirty BALB/c mice were randomly allocated to control, OVA, and MaR1 + OVA groups. Mice were sacrificed 24 hours after the end of the last challenge, and serum, bronchoalveolar lavage fluid (BALF), and lung tissue were collected for further analysis. Western blotting was used to measure the protein level of I

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cyclooxygenase 2; Disease Models, Animal; Docosahexaenoic Acids; Female; Intercellular Adhesion Molecule-1; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Signal Transduction

Previous reports have shown that pathogen-associated patterns (PAMPs) induce the production of interleukin (IL)-1β in macrophages. Moreover, studies using mouse models also suggest that chitin, which acts as a PAMP, induces adjuvant effects and eosinophilic infiltration in the lung. Thus, we investigated the effects of inhaled chitin in mouse models.. We developed mouse models of inhaled chitin particle-induced airway inflammation and steroid-resistant ovalbumin (OVA)-induced airway inflammation. Some experimental groups of mice were treated additionally with dexamethasone (DEX). Murine alveolar macrophages (AMs), which were purified from bronchoalveolar lavage (BAL) fluids, were incubated with chitin, and treated with or without DEX.. The numbers of total cells, AMs, lymphocytes, eosinophils, and neutrophils among BAL-derived cells, as well as the IL-1β levels in BAL fluids and the numbers of IL-1β-positive cells in lung, were significantly increased by chitin stimulation. Airway hyperresponsiveness (AHR) was aggravated in mice of the chitin inflammation model compared to control animals. The production of IL-1β was significantly increased in murine AMs by chitin treatment, but DEX administration did not inhibit this chitin-induced IL-1β production. Furthermore, in mouse models, DEX treatment inhibited the OVA-induced airway inflammation and AHR but not the airway inflammation and AHR induced by chitin or the combination of OVA and chitin.. These results suggest that inhaled chitin induces airway inflammation, AHR, and the production of IL-1β. Furthermore, our findings demonstrate for the first time that inhaled chitin induces steroid-resistant airway inflammation and AHR. Inhaled chitin may contribute to features of steroid-resistant asthma.

Topics: Administration, Inhalation; Animals; Asthma; Bronchoalveolar Lavage Fluid; Chitin; Dexamethasone; Disease Models, Animal; Drug Resistance; Glucocorticoids; Inflammation; Interleukin-1beta; Lung; Macrophages, Alveolar; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Pathogen-Associated Molecular Pattern Molecules; Respiratory Hypersensitivity

In this study, we investigated whether humidifier disinfectants (HDs) induce asthmatic airway inflammation in an animal model and compared the features of HD-induced inflammatory symptoms with ovalbumin (OVA)-induced allergic asthma. Mice were intratracheally instilled three times with either the control or 0.1, 0.3, or 0.5 mg/kg of polyhexamethylene guanidine phosphate (PHMG-P). To characterize asthmatic features, the following parameters were analyzed: (i) differential cell counts and cytokine expression in the bronchoalveolar lavage fluid (BALF); (ii) presence of mucus-producing goblet cells and pulmonary eosinophilic infiltration in the lungs; (iii) serum immunoglobulin levels; and (iv) airway hyperresponsiveness (AHR). RNA-Seq and bioinformatics tools were used to investigate whether PHMG-P altered asthma-related gene expression in lung tissues. The PHMG-P exposure groups showed higher peribronchial/perivascular inflammation, elevated goblet cell hyperplasia, and inhaled methacholine-induced airway resistance. Additionally, IL-13 and IL-17 in BALF were significantly increased in the PHMG-P exposure groups. However, there were no significant differences in total serum IgE and BALF IL-4 and IL-5 levels in the PHMG-P exposure groups compared to the control group. PHMG-P exposure modulated the expression of genes related to Th17 signaling pathways including the IL-17A, IL-23, and STAT3 signaling pathways, but not the Th2 signaling pathway. Altogether, our results suggest that repeated exposure to low does PHMG-P induces asthma-like symptoms and is thus a possible risk factor for developing asthma. The PHMG-P-induced asthmatic airway inflammation showed a different pattern from that found in typical allergic asthma and may be related to irritant-induced airway inflammation and hyperresponsiveness characterized by Th2-low, Th17-related, IgE-independent, and mixed granulocytic features.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Disinfectants; Female; Humidifiers; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; Th17 Cells

MicroRNA-181b (miR-181b) has been well noted with anti-inflammatory properties in several pathological conditions. It has also been suggested to be downregulated in patients with asthma. In this study, we explored the function of miR-181b in airway remodeling in asthmatic mice and the molecular mechanism. A mouse model with asthma was induced by ovalbumin (OVA) challenge, and miR-181b was found to be downregulated in lung tissues in the OVA-challenged mice. Overexpression of miR-181b was introduced in mice, after which the respiratory resistance, inflammatory infiltration, mucus production, and epithelial-mesenchymal transition (EMT) and fibrosis in mouse airway tissues were decreased. The integrated bioinformatics analysis suggested long non-coding RNA (lncRNA) TUG1 as a sponge for miR-181b. miR-181 directly targeted high mobility group box 1 (HMGB1) mRNA. HMGB1 was suggested to enhance activation of the nuclear factor kappa B (NF-κB) signaling. Further upregulation of lncRNA TUG1 blocked the protective functions of miR-181b in asthmatic mice. To conclude, this study evidenced that lncRNA TUG1 reinforces HMGB1 expression through sequestering microRNA-181b, which activates the NF-κB signaling pathway and promotes airway remodeling in asthmatic mice. This study may provide novel ideas in asthma management.

Topics: Airway Remodeling; Allergens; Animals; Asthma; Cells, Cultured; Disease Models, Animal; Female; HMGB1 Protein; Lung; Mice, Inbred BALB C; MicroRNAs; Mucus; NF-kappa B; Ovalbumin; RNA, Long Noncoding; Signal Transduction

Eosinophils are terminally differentiated cells derived from hematopoietic stem cells (HSCs) in the bone marrow. Several studies have confirmed the effective roles of eosinophils in asthmatic airway pathogenesis. However, their regulatory functions have not been well elucidated. Here, increased C-C chemokine ligand 6 (CCL6) in asthmatic mice and the human orthologs CCL15 and CCL23 that are highly expressed in asthma patients are described, which are mainly derived from eosinophils. Using Ccl6 knockout mice, further studies revealed CCL6-dependent allergic airway inflammation and committed eosinophilia in the bone marrow following ovalbumin (OVA) challenge and identified a CCL6-CCR1 regulatory axis in hematopoietic stem cells (HSCs). Eosinophil differentiation and airway inflammation were remarkably decreased by the specific CCR1 antagonist BX471. Thus, the study identifies that the CCL6-CCR1 axis is involved in the crosstalk between eosinophils and HSCs during the development of allergic airway inflammation, which also reveals a potential therapeutic strategy for targeting G protein-coupled receptors (GPCRs) for future clinical treatment of asthma.

Topics: Adolescent; Adult; Aged; Animals; Asthma; Bone Marrow; Cell Differentiation; Chemokines, CC; Eosinophils; Female; Healthy Volunteers; Hematopoietic Stem Cells; Humans; Hypersensitivity; Inflammation; Lung; Macrophage Inflammatory Proteins; Male; Mice; Mice, Knockout; Middle Aged; Ovalbumin; Phenylurea Compounds; Piperidines; Receptors, CCR1; Signal Transduction; Young Adult

Asthma is a common respiratory disease currently affecting more than 300 million worldwide and is characterized by airway inflammation, hyperreactivity, and remodeling. It is a heterogeneous disease consisting of corticosteroid-sensitive T-helper cell type 2-driven eosinophilic and corticosteroid-resistant, T-helper cell type 17-driven neutrophilic phenotypes. One pathway recently described to regulate asthma pathogenesis is cholesterol trafficking. Scavenger receptors, in particular SR-BI (scavenger receptor class B type I), are known to direct cellular cholesterol uptake and efflux. We recently defined SR-BI functions in pulmonary host defense; however, the function of SR-BI in asthma pathogenesis is unknown. To elucidate the role of SR-BI in allergic asthma, SR-BI-sufficient (SR-BI

Topics: Adrenal Insufficiency; Animals; Asthma; CD36 Antigens; Hypersensitivity; Inflammation; Interleukin-17; Lung; Male; Mice, Inbred C57BL; Neutrophils; Ovalbumin; Pyroglyphidae; Th17 Cells

In this study, immunoregulation and desensitization therapies were jointly applied in the treatment of asthma, in which chitosan (CS) nanoparticles were used. BALB/c mice were selected and mouse models of asthma were constructed. Mice were divided into 7 groups. A double-chamber plethysmograph, MTT, hematoxylin-eosin staining, and ELISA were used. The expression levels of IL-4 and IL-5 in lung tissue cells were detected. CS-BCG-PSN-OVA sustained-release vaccines significantly alleviated airway hyperresponsiveness (AHR) in asthmatic mice. The numbers of total lymphocytes and eosinophils in BALF were remarkably reduced. The expression levels of IL-4 and IL-5 in lung tissue cells of the treatment groups were dramatically decreased. CS-BCG-PSN-OVA was found in vitro to be able to inhibit OVA-induced T-cell proliferation and upregulate the proportion of CD4+CD25+Foxp3+ T cells. CS-BCG-PSN-OVA sustained-release vaccine could significantly attenuate AHR and airway inflammation in asthmatic mice. Thus, it has a promising application prospect for the treatment of bronchial asthma.

Topics: Animals; Asthma; BCG Vaccine; CD4-Positive T-Lymphocytes; Chitosan; Drug Liberation; Female; Inflammation; Interleukin-4; Interleukin-5; Lung; Mice, Inbred BALB C; Nanoparticles; Nucleic Acids; Ovalbumin; Polysaccharides

Autophagy plays an essential role in modulating asthma progression. MiR-20a-5p can regulate autophagy, but its effects on allergic asthma are still unclear. The aim of this study was to explore the potential of miR-20a-5p on autophagy-modulated airway remodeling and to reveal the underlying molecular mechanisms. We found that miR-20a-5p expression was markedly down-regulated in lung of ovalbumin (OVA)-induced mouse model with allergic asthma and in cells stimulated by OVA. Meanwhile, autophagy, apoptosis, fibrosis and inflammatory response were detected in pulmonary tissues from OVA-treated mice. Importantly, luciferase assays showed that ATG7 was a target of miR-20a-5p. We also found that miR-20a-5p over-expression markedly reduced ATG7, while its inhibition promoted ATG7 in cells. In addition, over-expressing miR-20a-5p in OVA-treated cells significantly decreased ATG7 expression levels, along with markedly reduced autophagy, apoptotic cell death, fibrosis and inflammatory response. These results were similar to the effects of autophagy inhibitor 3-Methyladenine (3-MA), indicating that miR-20a-5p was involved in autophagy-induced apoptosis, fibrosis and inflammation. In vivo experiments further demonstrated that miR-20a-5p over-expression was associated with ATG7 reduction in parallel with the alleviated airway remodeling in OVA-treated mice also through suppressing collagen accumulation, apoptosis and inflammation. Similarly, animal studies further confirmed that miR-20a-5p functioned as an autophagy inhibitor to mitigate allergic asthma development. Therefore, miR-20a-5p may be a promising biomarker and therapeutic target during asthma progression by regulating ATG7-modulated autophagy.

Topics: Allergens; Animals; Apoptosis; Asthma; Autophagy-Related Protein 7; Biomarkers; Disease Models, Animal; Down-Regulation; Female; Fibrosis; Inflammation; Lung; Mice, Inbred C57BL; MicroRNAs; Ovalbumin

Transient receptor potential ankyrin 1 (TRPA1) is a nonselective calcium-permeable ion channel highly expressed in the primary sensory neurons functioning as a polymodal sensor for exogenous and endogenous stimuli and has generated widespread interest as a target for inhibition due to its implication in neuropathic pain and respiratory disease. Herein, we describe the optimization of a series of potent, selective, and orally bioavailable TRPA1 small molecule antagonists, leading to the discovery of a novel tetrahydrofuran-based linker. Given the balance of physicochemical properties and strong

Topics: Animals; Asthma; CHO Cells; Cricetulus; Furans; Guinea Pigs; Humans; Inflammation; Ligands; Male; Molecular Structure; Ovalbumin; Oxadiazoles; Protein Binding; Purines; Rats, Sprague-Dawley; Structure-Activity Relationship; TRPA1 Cation Channel

Allergic asthma is one of the inflammatory diseases, which has become a major public health problem. Qu zhi qiao (QZQ), a dry and immature fruit of Citrus paradisi cv. Changshanhuyou, has various flavonoids with pharmacological properties. However, there is a knowledge gap on the pharmacological properties of QZQ on allergic asthma. Therefore, here, we explored the efficacy and mechanism of total flavonoids from QZQ (TFCH) on allergic asthma. We extracted and purified TFCH and conducted animal experiments using an Ovalbumin (OVA)-induced mice model. Bronchoalveolar lavage fluid and Swiss-Giemsa staining were used to count different inflammatory cells in allergic asthma mice. We conducted histopathology and immunohistochemistry to evaluate the changes in the lungs of allergic asthma mice. Moreover, we used ELISA assays to analyze chemokines and inflammatory cytokines. Furthermore, western blot analyses were conducted to elucidate the mechanism of TFCH on allergic asthma. We established that TFCH has anti-inflammatory effects and inhibits airway remodeling, providing a potential therapeutic strategy for allergic asthma.

Topics: Airway Remodeling; Animals; Asthma; Citrus paradisi; Drugs, Chinese Herbal; Female; Flavonoids; Fruit; Male; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinase Kinases; Ovalbumin; Signal Transduction; Smad2 Protein; Smad3 Protein

Jiawei Shengjiang Powder (JWSJP) is a classical Chinese medicinal formula, which has been widely applied in the treatment of asthma and complications for many years due to its curative effect.. To verify the effect of JWSJP in improving abnormal sperm motility caused by asthma and to explore its potential mechanism.. The active compounds of JWSJP were obtained from high performance liquid chromatography tandem mass spectrometry and the Traditional Chinese Medicine System Pharmacology. The key active components and targets of JWSJP were predicted based on network pharmacological analysis and bioinformatics research. Rats were randomly divided into normal, model and treatment groups. The rat model of allergic asthma was induced by intraperitoneal injection of ovalbumin solution. The experiment judged improvement of semen quality by evaluating sperm motility, and detected the expression of related proteins in testicular tissue of Sprague-Dawley rats by RT-qPCR and Western blot methods. Hematoxylin and eosin (HE) staining was used to observe the changes in testicular tissue structure in rats.. Through the analysis of network pharmacology and bioinformatics, it was found that beta-sitosterol, quercetin, gallic acid, pelargonidin and kaempferol were the key active components of Jiawei Shengjiang Powder. Tumor necrosis factor (TNF), interleukin-6 (IL-6) and insulin (INS) genes are crucial targets of JWSJP in the treatment of spermatogenic dysfunction caused by acute asthma. After 8 weeks of intervention, compared with the model group, the treatment group had significantly improved sperm motility (P < 0.05). There were significant differences in TNF, IL6, and INS proteins in the treatment group, and the HE staining of testicular tissue structure in the treatment group was significantly improved.. JWSJP can improve the abnormal sperm motility induced by asthma, and its mechanism may be related to the expression of related proteins and mRNA of TNF, IL6, and INS.

Topics: Animals; Asthenozoospermia; Asthma; Computational Biology; Disease Models, Animal; Drugs, Chinese Herbal; Male; Medicine, Chinese Traditional; Ovalbumin; Powders; Rats; Rats, Sprague-Dawley; Sperm Motility

Asthma is an inflammatory lung disease that affects more women than men in adulthood. Clinical evidence shows that hormonal fluctuation during the menstrual cycle and menopause are related to increased asthma severity in women. Considering that life expectancy has increased and that most women now undergo menopause, strategies to prevent the worsening of asthma symptoms are particularly important. A recent study from our group showed that re-exposure of ovariectomised allergic mice to antigen (ovalbumin) leads to an exacerbation of lung inflammation that is similar to clinical conditions. However, little is known about the role of probiotics in the prevention of asthma exacerbations during the menstrual cycle or menopause. Thus, our objective was to evaluate the effects of supplementation with kefir, a popular fermented dairy beverage, as a preventive strategy for modulating allergic disease. The results show that the preventive kefir administration decreases the influx of inflammatory cells in the airways and exacerbates the production of mucus and the interleukin 13 cytokine. Additionally, kefir changes macrophage polarisation by decreasing the number of M2 macrophages, as shown by RT-PCR assay. Thus, kefir is a functional food that potentially prevents allergic airway inflammation exacerbations in ovariectomised mice.

Topics: Animals; Asthma; Female; Fermentation; Humans; Interleukin-13; Kefir; Lactobacillales; Lung; Macrophage Activation; Macrophages; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics

Quinoline-3-carboxamides have been used to treat autoimmune/inflammatory diseases in humans because they inhibit the functions of S100 calcium-binding protein A9 (S100A9), which participates in the development of neutrophilic inflammation in asthmatics and in an animal model of neutrophilic asthma. However, the therapeutic effects of these chemicals have not been evaluated in asthma. The purpose of this study was to evaluate the effect of paquinimod, one of the quinoline-3-carboxamides, on a murine model of neutrophilic asthma.. Paquinimod was orally administered to 6-week-old C57BL/6 mice sensitized and challenged with ovalbumin (OVA)/complete Freund's adjuvant (CFA) and OVA. Lung inflammation and remodeling were evaluated using bronchoalveolar lavage (BAL) and histologic findings including goblet cell count. S100A9, caspase-1, IL-1. Paquinimod restored the enhancement of airway resistance and the increases in numbers of neutrophils and macrophages of BAL fluids and those of goblet cells in OVA/CFA mice toward the levels of sham-treated mice in a dose-dependent manner (0.1, 1, 10, and 25 mg/kg/day, p.o.). Concomitantly, p20 activated caspase-1, IL-1. These data indicate that paquinimod effectively inhibits neutrophilic inflammation and remodeling in the murine model of neutrophilic asthma, possibly via downregulation of IL-17, IFN-

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Freund's Adjuvant; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; Quinolines

This experimental study evaluated the anti-asthmatic capacity of the dihydroxyflavone chrysin in the settings of ovalbumin (OVA)-induced allergic inflammation.. The parameters that were used to assess the anti-asthmatic activity of chrysin included the specific airway resistance to histamine, the sensitivity to a chemically induced cough and the activity of chrysin on the ciliary beat frequency (CBF) of the respiratory epithelium. The anti-inflammatory potential was confirmed by the measurement of cytokine concentrations Th2 (IL-4, IL-5 and IL-13), Th1 (Granulocyte-macrophage colony-stimulating factor [GM-CSF], INF-γ and IL-12), leucocyte count in the bronchoalveolar lavage fluid (BALF) and growth factor TBF-β1 in lung homogenate.. Chronic administration of chrysin (30 mg/kg/day for 21 days) to OVA-sensitised guinea pigs showed bronchodilatory activity comparable to that of long-acting β 2 receptors agonist (LABA) salmeterol. Chrysin revealed antitussive efficiency but was not able to abolish the negative effect of OVA on CBF. Chrysin managed to ameliorate the progression of chronic airway inflammation by decreasing the count of eosinophils, lymphocytes and basophils, IL-5, L-13, GM-CSF, INF-γ in BALF, and TGF-β1 in lung homogenate.. The acquired results support the complex anti-asthmatic profile of chrysin. The flavone may represent an attractive compound for further studies concerning the prevention or treatment of asthma.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antitussive Agents; Asthma; Bronchoalveolar Lavage Fluid; Cough; Cytokines; Disease Models, Animal; Disease Progression; Flavonoids; Guinea Pigs; Inflammation; Male; Ovalbumin; Salmeterol Xinafoate

(1) Background: six mammalian ceramide synthases (CerS1-6) determine the acyl chain length of sphingolipids (SLs). Although ceramide levels are increased in murine allergic asthma models and in asthmatic patients, the precise role of SLs with specific chain lengths is still unclear. The role of CerS2, which mainly synthesizes C22-C24 ceramides, was investigated in immune responses elicited by airway inflammation using CerS2 null mice. (2) Methods: asthma was induced in wild type (WT) and CerS2 null mice with ovalbumin (OVA), and inflammatory cytokines and CD4 (cluster of differentiation 4)+ T helper (Th) cell profiles were analyzed. We also compared the functional capacity of CD4+ T cells isolated from WT and CerS2 null mice. (3) Results: CerS2 null mice exhibited milder symptoms and lower Th2 responses than WT mice after OVA exposure. CerS2 null CD4+ T cells showed impaired Th2 and increased Th17 responses with concomitant higher T cell receptor (TCR) signal strength after TCR stimulation. Notably, increased Th17 responses of CerS2 null CD4+ T cells appeared only in TCR-mediated, but not in TCR-independent, treatment. (4) Conclusions: altered Th2/Th17 immune response with higher TCR signal strength was observed in CerS2 null CD4+ T cells upon TCR stimulation. CerS2 and very-long chain SLs may be therapeutic targets for Th2-related diseases such as asthma.

Topics: Animals; Asthma; Mice; Mice, Knockout; Ovalbumin; Receptors, Antigen, T-Cell; Signal Transduction; Sphingosine N-Acyltransferase; Th17 Cells; Th2 Cells

Topics: Animals; Asthma; Complex Mixtures; GATA3 Transcription Factor; Hemiptera; Interleukin-17; Male; Mice; Mice, Inbred BALB C; Nuclear Receptor Subfamily 1, Group F, Member 3; Oleic Acid; Ovalbumin; Signal Transduction; Th2 Cells

Global industrialization not only improved the quality life of millions but also paved the way to solving many health problems. One among them is allergic asthma, which affects approximately 20% of the global population. Poor air quality is the major culprit in allergic asthma, which not only affects the individual's health, but also impairs his or her life quality and that of family members. Asthma is a chronic pulmonary inflammatory disease characterized by excess mucus production, airway hyperresponsiveness, and bronchoconstriction. Inhalation of corticosteroids, leukotriene modifiers, and β-adrenergic agonists is one treatment prescribed to control the symptoms of asthma, but there is still no effective cure. Phytochemicals such as carotenoids, phenolics, alkaloids, and nitrogen and organosulfur compounds are proven to possess immense pharmacological properties. Betalain is one such phytochemical present in plants of the order Caryophyllales. It is a water-soluble nitrogen-based pigment proven to possess antimicrobial, antioxidant, anti-inflammatory, hepatoprotective, antilipidemic, antidiabetic, and anticancer properties. We examined the curative potential of betalain against allergic asthma in a mouse model. Betalain treatment effectively decreased lung weight and infiltration of inflammatory cells in BAL fluid, and lowered IgE, eotaxin, and cytokine levels in asthma-induced mice. It also improved pulmonary mechanics and decreased oxidative stress and nitric oxide levels. Betalain significantly decreased gene expression of TGF-β and its downstream signaling Smad proteins. Lung histology confirmed that betalain protected the lung tissue of mice from ovalbumin-induced allergic asthma. Overall, our results show that betalain is a potent antiallergic drug that effectively protects mice from ovalbumin-induced allergic asthma. With further research, it can be prescribed as a treatment for asthma in humans.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antioxidants; Asthma; Betalains; Cytokines; Disease Models, Animal; Female; Immunoglobulin E; Lung; Mice, Inbred BALB C; Ovalbumin; Signal Transduction; Smad Proteins; Transforming Growth Factor beta1

T cell immunoglobulin and mucin domain-containing molecule-3 (Tim-3) has been described as a transmembrane protein, expressed on the surface of various T cells as well as different cells of innate immunity. It has since been associated with Th1 mediated autoimmune diseases and transplantation tolerance studies, thereby indicating a possible role of this receptor in counter-regulation of Th2 immune responses. In the present study we therefore directly examined the role of Tim-3 in allergic inflammation and respiratory tolerance. First, Tim-3-/- mice and wild type controls were immunized and challenged with the model allergen ovalbumin (OVA) to induce an asthma-like phenotype. Analysis of cell numbers and distribution in the bronchoalveolar lavage (BAL) fluid as well as lung histology in H&E stained lung sections demonstrated a comparable degree of eosinophilic inflammation in both mouse strains. Th2 cytokine production in restimulated cell culture supernatants and serum IgE and IgG levels were equally increased in both genotypes. In addition, cell proliferation and the distribution of different T cell subsets were comparable. Moreover, analysis of both mouse strains in our respiratory tolerance model, where mucosal application of the model allergen before immunization, prevents the development of an asthma-like phenotype, revealed no differences in any of the parameters mentioned above. The current study demonstrates that Tim-3 is dispensable not only for the development of allergic inflammation but also for induction of respiratory tolerance in mice in an OVA-based model.

Topics: Allergens; Animals; Asthma; Cytokines; Female; Hepatitis A Virus Cellular Receptor 2; Immune Tolerance; Immunization; Immunoglobulin E; Inflammation; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Respiratory Tract Diseases; Th2 Cells

In traditional Chinese medicine (TCM), the leaf of Elaeagnus pungens Thunb. (Family Elaeagnaceae) is a herb documented as an antiasthmatic remedy to treat the severe asthma, bronchitis and other respiratory diseases in the early material medica "Bencao Gangmu" (Ming dynasty, about 442 years ago).. This work is purposed to investigate the pharmacological effects and mechanism of total flavonoids from Elaeagnus pungens leaves (FLA) on asthma in vivo and vitro.. Female BALB/c mice were sensitized by intraperitoneal injection of OVA with aluminum hydroxide and intranasal challenged with OVA. After treatment with FLA (10, 20 mg/kg p.o.), the behaviors of mice were observed by score evaluation. Enumeration of total cells and OVA-specific IgE assay in the blood were measured as well as enumeration of total cells and cytokines assay in the BALF. Furthermore, histopathological analysis was performed by HE staining. The in vitro relaxing action on muscle force of FLA (0.0316-10.0 mg/ml) was evaluated using isometric tension in tracheal rings, and VDLCC currents were recorded to explore the relaxation mechanism in the isolated tracheal rings and mouse ASM cells, respectively. In vitro anti-inflammatory actions were assessed with LPS-stimulated RAW 264.7 macrophages. The production of inflammatory mediators and MAPK signaling pathway was estimated using ELISA and Western blotting analysis, respectively.. The high dose of flavones from E. pungens leaf (20 mg/kg) can significantly improve the symptom of asthma breakout and relieve the lung swelling. FLA treatment decreased eosinophils and leukocytes numbers in blood and BLAF with a dosedependent manner. Furthermore, the inhibiting effect of FLA on the level of Ig E and inflammatory-related cytokines including TNF-α, IL-5 showed dose-independent. FLA relaxed high K + -induced contraction in a dose-dependent manner. The maximal relaxation produced by FLA was 99.7% (IC 50 = 0.46 mg/ml). The whole-cell VDLCC currents were abolished by FLA (3.16 mg/ml) and FLA significantly decreased the maximal amplitude of VDLCCs. No cytotoxic effect of FLA was observed in RAW264.7 cells under the tested concentrations (1-300 μg/mL). The increased IL-6 and NO by the stimulation of LPS in RAW264.7 cells were significantly inhibited by FLA in the dosedependent manner. Treatment with LPS in the presence of FLA, LPS-induced phosphorylation of ERK1/2 and JNK was inhibited in the macrophages.. FLA from Elaeagnus pungens leaf can alleviate the inflammation symptom via reducing the eosinophils and leukocytes numbers as well as the production of pro-inflammatory cytokines. This anti-inflammatory effect is related to the modulation of the MAPK signaling pathway. FLA can relax the precontracted TRs by blocking the VDLCCs, which interrupts extracellular Ca 2+ influx and inhibit the rise of [Ca 2+ ]i. It strongly suggests that these flavonoids components are the substances basis of Elaeagnus pungens leaves for allergic action, bronchospasm and inflammation in asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Calcium Channels, L-Type; Cytokines; Disease Models, Animal; Elaeagnaceae; Female; Flavonoids; Immunoglobulin E; Inflammation; Lipopolysaccharides; Macrophages; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Muscle Relaxation; Ovalbumin; Plant Extracts; Plant Leaves; RAW 264.7 Cells; Trachea

Asthma is a respiratory disease with a dramatically increasing incidence globally. The present study explored the roles of S-phase kinase-associated protein 2 (SKP2) and forkhead box O3 (FOXO3) in asthma and their involvement in the Krüppel-like factor 15-lipoprotein receptor-related protein 5 (KLF15-LRP5) axis. SKP2 expression in patients with asthma and OVA-induced asthmatic Sprague Dawley rats was detected by reverse transcription quantitative PCR and Western blot assays. Alterations in SKP2 and LRP5 expression were evaluated in OVA-induced asthmatic rats, followed by measurement of inflammatory cytokines using ELISA and airway resistance using a methacholine challenge test. We applied TGF-β1 to establish the airway smooth muscle cell (ASMC) proliferation model of asthma. The FOXO3 ubiquitination and changes in cell biological behaviors were detected using immunoprecipitation, MTT, and Annexin V/propidium iodide assays. Flow cytometry was adopted to detect cell cycle, and ELISA was used to measure the concentrations of IL-4, IL-5, IL-13, and IgE in rat bronchoalveolar lavage fluid. SKP2 was highly expressed and FOXO3 was poorly expressed in patients with asthma and in OVA-induced asthmatic rats. SKP2 silencing decreased IL-4, IL-5, IL-13, and IgE expression in rat bronchoalveolar lavage fluid, whereas SKP2 enhanced FOXO3 ubiquitination to upregulate KLF15, which bound to the LRP5 promoter in TGF-β1-induced ASMCs and increased LRP5 expression. SKP2 enhanced airway hyperresponsiveness and inflammation in the OVA-induced rat model and augmented TGF-β1-induced ASMC proliferation by inhibiting the FOXO3/KLF15/LRP5 axis. Additionally, overexpressed SKP2 resulted in reduced numbers of ASMCs in the G1 phase but increased numbers in the G2/M phase. Collectively, we show that SKP2 promotes FOXO3 ubiquitination to suppress the KLF15-LRP5 axis, thereby exacerbating asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Case-Control Studies; Cell Proliferation; Cells, Cultured; Disease Models, Animal; Female; Forkhead Box Protein O3; Humans; Male; Myocytes, Smooth Muscle; Ovalbumin; Rats; Rats, Sprague-Dawley; S-Phase Kinase-Associated Proteins; Signal Transduction; Trachea; Transforming Growth Factor beta1; Ubiquitination

Asthma is one of the most common noninfectious chronic diseases characterized by type II inflammation. This study aimed to investigate the effects of molecular hydrogen on the pathogenesis of asthma.. OVA sensitized asthma mouse model and house dust mite treated 16HBE cellular model were established and hydrogen/oxygen mixture was used to treat asthmatic mice and 16HBE cells. Serum and BALF cytokines were measured with specific ELISA assays. E-cadherin and ZO-1 were detected by immunohistochemical staining and expression of caspase 3 and 9, NF-κB, IL-33 and ST2 was assessed by quantitative real-time PCR, western blot and/or immunofluorescence. IL-33 promoter activity was analyzed by dual-luciferase assay. ILC2 population was assayed by flow cytometry and differentially expressed miRNAs were detected using miRNA array.. Serum and BALF levels of IL-33 and other alarmin and type II cytokines were greatly increased by OVA and inhibited by H. These data demonstrated that H

Topics: Allergens; Animals; Anti-Asthmatic Agents; Apoptosis; Asthma; Bronchoalveolar Lavage Fluid; Cell Line; Cytokines; Epithelial Cells; Female; Humans; Hydrogen; Interleukin-1 Receptor-Like 1 Protein; Mice, Inbred ICR; MicroRNAs; Ovalbumin; Pyroglyphidae; Respiratory Mucosa

Asthma-induced pulmonary fibrosis (PF) is an important public health concern that has few treatment options given its poorly understood etiology; however, the epithelial to mesenchymal transition (EMT) of pulmonary epithelial cells has been implicated to play an important role in inducing PF. Although previous studies have found atractylon (Atr) to have anti-inflammatory effects, whether Atr has anti-PF abilities remains unknown. The purpose of the current study was to validate the protective efficiency of Atr in both an animal model of ovalbumin (OVA)-induced asthma and an EMT model induced by transforming growth factor-β1 (TGF-β1) using TC-1 cells. The results of this study revealed that Atr treatment suppressed OVA-induced PF via fibrosis-related protein expression. Atr treatment suppressed OVA-induced circRNA-0000981 and TGFBR2 expression but promoted miR-211-5p expression. In vivo studies revealed that Atr suppressed TGF-β1-induced EMT and fibrosis-related protein expression via suppressing circRNA-0000981 and TGFBR2 expression. The results also suggested that the downregulation of circRNA-0000981 expression suppressed TGFBR2 by sponging miR-211-5p, which was validated by a luciferase reporter assay. Collectively, the findings of the present study suggest that Atr treatment attenuates PF by regulating the mmu_circ_0000981/miR-211-5p/TGFBR2 axis in an OVA-induced asthma mouse model.

Topics: Animals; Asthma; Cell Line; Male; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Pulmonary Fibrosis; Receptor, Transforming Growth Factor-beta Type II; RNA, Circular; Sesquiterpenes; Treatment Outcome

The combined allergic rhinitis and asthma syndrome (CARAS) is a chronic airway inflammation of allergic individuals, with a type 2 immune response. Pharmacotherapy is based on drugs with relevant side effects. Thus, the goal of this study was to evaluate the synthetic alkaloid, MHTP in the experimental model of CARAS. Therefore, BALB/c mice were ovalbumin (OVA) -sensitized and -challenged and treated with MHTP by intranasal or oral routes. Treated animals showed a decrease (p < 0.05) of sneezing, nasal rubbings, and histamine nasal hyperactivity. Besides, MHTP presented binding energy and favorable interaction for adequate anchoring in the histamine H1 receptor. MHTP treatment inhibited the eosinophil migration into the nasal (NALF) and the bronchoalveolar (BALF) fluids. Histological analysis showed that the alkaloid decreased the inflammatory cells in the subepithelial and perivascular regions of nasal tissue and in the peribronchiolar and perivascular regions of lung tissue. The MHTP treatment also reduced the pulmonary hyperactivity by decreasing the smooth muscle layer hypertrophy and the collagen fiber deposition in the extracellular matrix. The immunomodulatory effect of the alkaloid was due to the decrease of cytokines like IL-5 and IL-17A (type 2 and 3), TSLP (epithelial), and the immunoregulatory cytokine, TGF-β. These MHTP effects on granulocytes were dependent on the p38/ERK1/2 MAP kinase signaling pathway axis. Indeed, the synthetic alkaloid reduced the frequency of activation of both kinases independent of the NF-κB (p65) pathway indicating that the molecule shut down the intracellular transduction signals underlie the cytokine gene transcription.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Disease Models, Animal; Female; Humans; MAP Kinase Signaling System; Mice; Mice, Inbred BALB C; Molecular Docking Simulation; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Receptors, Histamine H1; Rhinitis, Allergic; Tetrahydroisoquinolines

Sophocarpine (SPC) as a quinolizidine alkaloid displays powerful effects on inflammatory diseases through regulating multiple targets. Asthma is a complex heterogeneous and inflammatory disease with an increasing incidence worldwide. Here we established a mice asthma model and investigated the effect of SPC. Mice induced by ovalbumin (OVA) exhibits exacerbated Th1/Th2 immune imbalance and allergic lung inflammation. SPC treatment regulated Th1/Th2 cytokines production (IL-4, IL-5 and INF-γ) in BALF, reduced IgE level in serum, inhibited inflammatory cell infiltration, and improved the lung tissue pathology. Proteomic results showed that 5064 proteins in lung tissue were detected and among them 223 preliminary therapeutic targets of SPC were selected. Subsequently, excluding non-human genes, 109 targets with established crystal structures were harvested. Meanwhile, the molecular docking results showed that the binding energy of 87 targets with SPC was varied from -9.72 kcal/mol to 227.16 kcal/mol. Further, SPC suppressed arrb2, anxa1, myd88 and sphk1 expression and activated p-stat1. All of the five targets based on the screened results of proteomics and molecular docking are critical in allergic asthma. Thus, our data revealed that SPC alleviated bronchial asthma via targeting multi-targets.

Topics: Alkaloids; Allergens; Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Disease Models, Animal; Female; Humans; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Molecular Docking Simulation; Molecular Targeted Therapy; Ovalbumin; Proteome; Th2 Cells

Asthma is an inflammatory airway disease wherein bronchoconstriction, airway inflammation, and airway obstruction during asthma attacks are the main problems. It is recognized that imbalance of Th1/Th2 and Th17/Treg is a critical factor in asthma pathogenesis. Manipulation of these with signaling molecules such as mTOR, PI3K, Akt, and MyD88 can control asthma. Mouse model of allergic asthma was produced and treated with ketamine, metformin, metformin and ketamine, triciribine, LY294002, and torin2. MCh challenge test, BALf's Eos Count, the IL-4, 5, INF-γ, eicosanoid, total IgE levels were determined. The MUC5a, Foxp3, RORγt, PI3K, mTOR, Akt, PU.1, and MyD88 gene expressions and histopathology study were done. Asthma groups that were treated with all six components had reduced Penh value, total IgE, IL-4 and IL-5 levels, MUC5a, RORγt, MyD88 and mTOR expression, goblet cell hyperplasia, and mucus hyper-secretion. The eosinophil percentage and Cys-LT level were decreased by metformin and ketamine, triciribine, LY294002, and torin2. The level of IFN-γ was increased in triciribine, LY294002, and torin2. Metformin, metformin and ketamine, triciribine, LY294002, and torin2 reduced Akt and PI3K expression, peribronchial and perivascular inflammation, and increased expression of Foxp3. Torin2 had an effect on PU.1 expression. Inhibition of PI3K/AKT/mTOR and TLR4/MyD88/NF-κB signaling with targeted molecules can attenuate asthma pathology and play an important role in airways protection.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Chromones; Female; Metformin; Mice; Mice, Inbred BALB C; Morpholines; Myeloid Differentiation Factor 88; Naphthyridines; NF-kappa B; Ovalbumin; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Signal Transduction; Toll-Like Receptor 4; TOR Serine-Threonine Kinases

Asthma is a chronic airway inflammation that caused by many factors. The voltage-gated proton channel Hv1 has been proposed to extrude excessive protons produced by NADPH oxidase (NOX) from cytosol to maintain its activity during respiratory bursts. Here, we showed that loss of Hv1 aggravates ovalbumin (OVA)-induced allergic lung asthma in mice. The numbers of total cells, eosinophils and neutrophils in bronchoalveolar lavage fluid (BALF) of Hv1-deficiency (KO) mice are obviously increased after OVA challenge compared with that of wild-type (WT) mice. Histopathological staining reveals that Hv1-deficiency aggravates OVA-induced inflammatory cell infiltration and goblet cell hyperplasia in lung tissues. The expression of IL-4, IL-5 and IL-13 are markedly increased in lung tissues of OVA-challenged KO mice compared with that of WT mice. Furthermore, the expression levels of NOX2, NOX4 and DUOX1 are dramatically increased, while the expression levels of SOD2 and catalase are significantly reduced in lung tissues of OVA-challenged KO mice compared with that of WT mice. The production of ROS in lung tissues of KO mice is significantly higher than that of WT mice after OVA challenge. Our data suggest that Hv1-deficiency might aggravate the development of allergic asthma through increasing ROS production.

Topics: Acetylcysteine; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Goblet Cells; Hyperplasia; Ion Channels; Mice, Knockout; NADPH Oxidases; Ovalbumin; Reactive Oxygen Species; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Th2 Cells

The occurrence of asthma is closely related to environmental factors such as cigarette smoke (CS), one of the common risk factors. Environmental stimuli have the potential to activate transient receptor potential ankyrin 1 (TRPA1) and cause or aggravate asthma. The destruction of tight junctions (TJs) between airway epithelial cells by environmental stimuli in asthma has been researched. It is worth exploring whether CS can injury TJs and aggravate asthma by activating TRPA1. The objective of this study was to investigate the aggravation of CS on ovalbumin (OVA)-induced asthma related phenotypes and TJs expression in mice, and to explore the relationship between TRPA1 and the expression of TJs protein. Female wild type (WT) C57BL/6 mice, induced by OVA, CS and OVA plus CS (OVA + CS) respectively, were used to establish a 42-day asthma model, and mice with TRPA1 knockout (TRPA1

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Claudins; Disease Models, Animal; Enzyme Activation; Interleukin-13; Interleukin-4; Interleukin-5; Mice; Mice, Inbred C57BL; Mice, Knockout; Nicotiana; Occludin; Ovalbumin; Smoke; Smoking; Tight Junctions; TRPA1 Cation Channel; Zonula Occludens-1 Protein

Asthma is a complex disorder characterized by chronic inflammation of the airways. We aimed to investigate the role of Atractylenolide III (ATL III) in ovalbumin (OVA)-induced mouse asthma. Asthma was induced to BALB/c mice by sensitization with intraperitoneal injection of OVA, followed by treatment with ATL III. Pathological changes in lung tissue were examined by hematoxylin/ eosin and sirius red staining. The levels of inflammation- and oxidative stress-related factors in the bronchoalveolar lavage fluid (BALF) were monitored using kits. Additionally, the contents of inflammatory cells including macrophages, lymphocytes, eosinophils and neutrophils in BALF were counted. The expression of signal transducer and activator of transcription 3 (STAT3) was tested using Western blotting and immunohistochemistry assay. Results revealed that ATL III markedly attenuated OVA-induced pathological injury of lung tissues in mice. Furthermore, ATL III controlled the cytokines production and balanced the oxidative stress condition, which was exhibited by the reduced levels of inflammation- and oxidative stress-related factors. Moreover, mice in ATL III-treated groups presented less inflammatory cells in BALF and ATL III largely inhibited STAT3 expression in lung tissues. Taken together, ATL III alleviates inflammation, oxidative stress and is associated with changes in pulmonary functions in a mouse asthma model through inhibiting STAT3.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Inflammation; Lactones; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Sesquiterpenes

2,6-Bis-(4-hydroxyl-3-methoxybenzylidine) cyclohexanone (BHMC), a synthetic curcuminoid analogue, has been shown to exhibit anti-inflammatory properties in cellular models of inflammation and improve the survival of mice from lethal sepsis. We further evaluated the therapeutic effect of BHMC on acute airway inflammation in a mouse model of allergic asthma. Mice were sensitized and challenged with ovalbumin (OVA), followed by intraperitoneal administration of 0.1, 1, and 10 mg/kg of BHMC. Bronchoalveolar lavage fluid, blood, and lung samples were collected, and the respiratory function was measured. OVA sensitization and challenge increased airway hyperresponsiveness (AHR) and pulmonary inflammation. All three doses of BHMC (0.1-10 mg/kg) significantly reduced the number of eosinophils, lymphocytes, macrophages, and neutrophils, as well as the levels of Th2 cytokines (IL-4, IL-5 and IL-13) in bronchoalveolar lavage fluid (BALF) as compared to OVA-challenged mice. However, serum level of IgE was not affected. All three doses of BHMC (0.1-10 mg/kg) were effective in suppressing the infiltration of inflammatory cells at the peribronchial and perivascular regions, with the greatest effect observed at 1 mg/kg which was comparable to dexamethasone. Goblet cell hyperplasia was inhibited by 1 and 10 mg/kg of BHMC, while the lowest dose (0.1 mg/kg) had no significant inhibitory effect. These findings demonstrate that BHMC, a synthetic nonsteroidal small molecule, ameliorates acute airway inflammation associated with allergic asthma, primarily by suppressing the release of inflammatory mediators and goblet cell hyperplasia to a lesser extent in acute airway inflammation of allergic asthma.

Topics: Acute Disease; Animals; Asthma; Bronchial Hyperreactivity; Curcumin; Cyclohexanones; Cytokines; Goblet Cells; Immunoglobulin E; Leukocytes; Male; Mice; Mice, Inbred BALB C; Ovalbumin

Allergic asthma is a common airway inflammatory disorder with increasing morbidity and mortality worldwide. Gentiopicroside (GPS) is a secoiridoid glycoside compound that exhibits anti-inflammatory property. However, the effect of GPS on allergic asthma has not been reported yet. In this study, we investigated the role of GPS in a mouse model of ovalbumin (OVA)-induced allergic asthma and explored its potential mechanism. Mice were sensitized with OVA and gavaged with 20, 40, or 80 mg/kg GPS. Administration of GPS decreased lung wet-to-dry weight ratio. Histological analysis of H&E and PAS staining showed that GPS treatment alleviated inflammatory cell infiltration and goblet cell hyperplasia in lung tissue of OVA-sensitized mice. Moreover, GPS inhibited the recruitment of inflammatory cells including total cells, macrophages, eosinophils, lymphocytes and neutrophils and the secretion of T helper type 2 (Th2) cytokines (interleukin (IL)-4, IL-5 and IL-13) in bronchoalveolar lavage fluid (BALF) of OVA-sensitized mice in a dose dependent manner. The levels of OVA-specific immunoglobulin E (IgE) and pro-inflammatory tumor necrosis factor (TNF)-α were also attenuated by GPS treatment. Interestingly, GPS upregulated the expression of silent information regulator 1 (SIRT1) while downregulated the expression of acetyl-nuclear factor kappa B (NF-κB) p65 in lung tissue of OVA-sensitized mice. Furthermore, treatment with an SIRT1 inhibitor (EX-527) partially abolished the inhibitory effect of GPS on OVA-induced airway inflammation, suggesting that the anti-inflammation of GPS might be achieved through regulating SIRT1/NF-κB p65 signaling pathway. These findings indicate that GPS might be a novel drug candidate in the treatment of allergic asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Inflammation; Iridoid Glucosides; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Signal Transduction; Sirtuin 1

Asthma is characterized by airway remodeling. Glucocorticoid induced transcript 1 (GLCCI1) was reported to be associated with the development of asthma, while its exact mechanism is still not clear. In our study, ovalbumin (OVA) combined with aluminum hydroxide were used to establish asthmatic mouse model. ELISA assay was fulfilled to ensure the concentration of inflammatory factors in both bronchoalveolar lavage fluid and serum. The pathological changes and collagen deposition in lung tissues were analyzed using H&E staining and Masson staining, respectively. The expression of proteins was measured using western blot, and the expression of GLCCI1 mRNA was ensured by qRT-PCR. Here, we demonstrated that OVA-induced inflammation, lung structural remodeling and collagen deposition in asthmatic mice was notably improved by hydroprednisone treatment or GLCCI1 overexpressing. The expression of GLCCI1 was decreased, while IL-13, periostin and TGF-β1 were increased in the lung tissue of asthmatic mice. Importantly, upregulation of GLCCI1 suppressed the expression of IL-13, periostin and TGF-β1, phosphorylation of Smad2 and Smad3, and extracellular matrix (ECM) deposition-related proteins expression. IL-13-induced upregulation of periostin and TGF-β1 expression, phosphorylation of Smad2 and Smad3, and ECM deposition in airway epithelial cells (AECs) was repressed by GLCCI1 increasing. Furthermore, our results showed that overexpression of GLCCI1 repressed the effect of IL-13 on AECs via inhibiting periostin expression. Overall, our data revealed that GLCCI1 limited the airway remodeling in mice with asthma through inhibiting IL-13/periostin/TGF-β1 signaling pathway. Our data provided a novel target for asthma treatment.

Topics: Airway Remodeling; Aluminum Hydroxide; Animals; Asthma; Cell Adhesion Molecules; Disease Models, Animal; Female; Humans; Interleukin-13; Lung; Mice; Ovalbumin; Prednisone; Receptors, Glucocorticoid; Signal Transduction; Transforming Growth Factor beta1

Asthma is a well-known bronchial disease that causes bronchial inflammation, narrowing of the bronchial tubes, and bronchial mucus secretion, leading to bronchial blockade. In this study, we investigated the association between phosphodiesterase (PDE), specifically PDE1, and asthma using 3-isobutyl-1-methylxanthine (IBMX; a non-specific PDE inhibitor) and vinpocetine (Vinp; a PDE1 inhibitor). Balb/c mice were randomized to five treatment groups: control, ovalbumin (OVA), OVA + IBMX, OVA + Vinp, and OVA + dexamethasone (Dex). All mice were sensitized and challenged with OVA, except for the control group. IBMX, Vinp, or Dex was intraperitoneally administered 1 h before the challenge. Vinp treatment significantly inhibited the increase in airway hyper-responsiveness (P<0.001) and reduced the number of inflammatory cells, particularly eosinophils, in the lungs (P<0.01). It also ameliorated the damage to the bronchi and alveoli and decreased the OVA-specific IgE levels in serum, an indicator of allergic inflammation increased by OVA (P<0.05). Furthermore, the increase in interleukin-13, a known Th2 cytokine, was significantly decreased by Vinp (P<0.05), and Vinp regulated the release and mRNA expression of macrophage inflammatory protein-1β (MIP-1β) increased by OVA (P<0.05). Taken together, these results suggest that PDE1 is associated with allergic lung inflammation induced by OVA. Thus, PDE1 inhibitors can be a promising therapeutic target for the treatment of asthma.

Topics: 1-Methyl-3-isobutylxanthine; Animals; Anti-Inflammatory Agents; Asthma; Chemokine CCL4; Dexamethasone; Disease Models, Animal; Down-Regulation; Gene Expression Regulation; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Random Allocation; Vinca Alkaloids

Topics: Administration, Inhalation; Aerosols; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Lung; Mice; Mice, Inbred BALB C; Mycobacteriaceae; Ovalbumin

Studying the proteomes of tissue-derived extracellular vesicles (EVs) can lead to the identification of biomarkers of disease and can provide a better understanding of cell-to-cell communication in both healthy and diseased tissue. The aim of this study was to apply our previously established tissue-derived EV isolation protocol to mouse lungs in order to determine the changes in the proteomes of lung tissue-derived EVs during allergen-induced eosinophilic airway inflammation. A mouse model for allergic airway inflammation was used by sensitizing the mice intraperitoneal with ovalbumin (OVA), and one week after the final sensitization, the mice were challenged intranasal with OVA or PBS. The animals were sacrificed 24 h after the final challenge, and their lungs were removed and sliced into smaller pieces that were incubated in culture media with DNase I and Collagenase D for 30 min at 37 °C. Vesicles were isolated from the medium by ultracentrifugation and bottom-loaded iodixanol density cushions, and the proteomes were determined using quantitative mass spectrometry. More EVs were present in the lungs of the OVA-challenged mice compared to the PBS-challenged control mice. In total, 4510 proteins were quantified in all samples. Among them, over 1000 proteins were significantly altered (fold change >2), with 614 proteins being increased and 425 proteins being decreased in the EVs from OVA-challenged mice compared to EVs from PBS-challenged animals. The associated cellular components and biological processes were analyzed for the altered EV proteins, and the proteins enriched during allergen-induced airway inflammation were mainly associated with gene ontology (GO) terms related to immune responses. In conclusion, EVs can be isolated from mouse lung tissue, and the EVs' proteomes undergo changes in response to allergen-induced airway inflammation. This suggests that the composition of lung-derived EVs is altered in diseases associated with inflammation of the lung, which may have implications in type-2 driven eosinophilic asthma pathogenesis.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Extracellular Vesicles; Gene Ontology; Lung; Male; Mice; Mice, Inbred C57BL; Mitochondria; Nanoparticles; Ovalbumin; Proteome; Pulmonary Eosinophilia; Respiratory Hypersensitivity

Tregitopes (T regulatory epitopes) are IgG-derived peptides with high affinity to major histocompatibility complex class II (MHCII), that are known to promote tolerance by activating T regulatory cell (Treg) activity. Here we characterized the effect of IgG Tregitopes in a well-established murine model of allergic asthma, demonstrating

Topics: Adoptive Transfer; Animals; Animals, Genetically Modified; Anti-Asthmatic Agents; Antigens, Plant; Asthma; Bronchoconstriction; Cells, Cultured; Cytokines; Disease Models, Animal; Epitopes, T-Lymphocyte; Humans; Immunoglobulin Fab Fragments; Immunoglobulin Fc Fragments; Inflammation Mediators; Lung; Lymphocyte Activation; Mice, Inbred C57BL; Ovalbumin; Plant Extracts; T-Lymphocytes, Regulatory

Asthma is a chronic airway disease that causes excessive inflammation, oxidative stress, mucus production and bronchial epithelial cell apoptosis. Fructose-1,6-bisphosphatase (Fbp1) is one of the rate-limiting enzymes in gluconeogenesis and plays a critical role in several cancers. However, its role in inflammatory diseases, such as asthma, is unclear. Here, we examined the expression, function and mechanism of action of Fbp1 in asthma. Gene Expression Omnibus (GEO) data sets revealed that Fbp1 was overexpressed in a murine model of asthma and in interleukin (IL)-4- or IL-13-stimulated bronchial epithelial cells. We confirmed the findings in an animal model as well as Beas-2B and 16HBE cells. In vitro investigations revealed that silencing of Fbp1 reduced apoptosis and the proportion of cells in the G2/M phase, whereas overexpression led to increases. Fbp1 knock-down inhibited oxidative stress by activating the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, whereas Fbp1 overexpression aggravated oxidative stress by suppressingthe Nrf2 pathway. Moreover, the Nrf2 pathway inhibitor ML385 reversed the changes caused by Fbp1 inhibition in Beas-2B and 16HBE cells. Collectively, our data indicate that Fbp1 aggravates oxidative stress-induced apoptosis by suppressing Nrf2 signalling, substantiating its potential as a novel therapeutic target in asthma.

Topics: Animals; Asthma; Female; Fructose-Bisphosphatase; Gene Expression Regulation; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; Ovalbumin; Oxidative Stress

The objective of the study is to evaluate the protective effects of human mesenchymal stem cells (hMSCs) modified with miR-138-5p inhibitor against the allergic rhinitis and asthma syndrome (ARAS). MiR-138-5p or negative control was transfected into hMSCs, and fluorescence-activated cell sorting was used to evaluate hMSC surface markers. Quantitative real-time PCR (qRT-PCR) was used to evaluate miR-138-5p, SIRT1, caspase-3, IL-6, IL-1β and TNF-α levels after TNF-α and IL-6 stimulations. hMSCs with or without miR-138-5p inhibition was intranasally administered into ARAS mice (n = 10 each group), followed by monitoring sneezing and nasal rubbing events to evaluate the allergic symptoms. Histamine, ovalbumin-specific IgE, IgG2a, IgG1 and LTC4 release were monitored in the serum and nasal lavage fluid using enzyme-linked immunosorbent assay. Expression of SIRT1 and HMGB1/TLR4 pathway in nasal mucosa was assessed. After miR-138-5p inhibitor transfection, the hMSC lineage was preserved. Binding between SIRT1 and miR-138-4p was observed, and miR-138-5p inhibition led to upregulation of SIRT1. Inhibition of miR-138-5p led to attenuated inflammatory responses of hMSCs upon TNF-α and IL-6 stimulation, and allergic symptoms in mice, as well as histamine and ovalbumin-specific IgG release. hMSCs with miR-138-5p inhibition showed characteristics of activated SIRT1 and inhibited HMGB1/TLR4 pathway. Inhibition of miR-138-5p in hMSCs enhanced its effects in attenuating inflammatory responses and allergic reaction in the ARAS model, which is presumably regulated by SIRT1 and the HMGB1/TLR4 pathway.

Topics: Animals; Apoptosis; Asthma; Cell Proliferation; Cells, Cultured; Cytokines; Female; HMGB1 Protein; Humans; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Protective Agents; Rhinitis, Allergic; Sirtuin 1

Typical murine models of allergic inflammation are induced by the combination of ovalbumin and aluminum hydroxide. However, accumulating evidence indicates that, in models of asthma and atopic dermatitis, allergic inflammation can be generated in the absence of aluminum hydroxide. Moreover, co-administration of Staphylococcus aureus enterotoxin B with ovalbumin can enhance inflammation. The objective of this study was to establish a rapid and mast cell-dependent murine model of allergic inflammation by inducing allergic peritonitis using ovalbumin and S. aureus enterotoxin B. Allergic peritonitis was induced in C57BL/6 mice by subcutaneous sensitization and intraperitoneal challenge with ovalbumin and S. aureus enterotoxin B. Disease characteristics were assessed by flow cytometry, enzyme-linked immunosorbent assay (ELISA), trypan blue exclusion and colorimetric assays. The time-course of the allergic peritonitis revealed a peak of peritoneal inflammation 48 h after challenge, as assessed by total cells and eosinophil counts. The decrease of cell numbers started 96 h post-challenge, with complete clearance within 168 h. Moreover, significantly higher levels of tryptase and increased vascular permeability were found 30 min following challenge. Allergic inflammation induction by ovalbumin and S. aureus enterotoxin B was impaired in mast cell-deficient mice and partially restored by mice reconstitution with bone marrow-derived mast cells, indicating the mast cell role in this model. We present a novel model of allergic peritonitis that is mast cell-dependent, simple and robust. Moreover, the use of S. aureus enterotoxin B better resembles human allergic inflammation, which is known to be characterized by the colonization of S. aureus.

Topics: Animals; Asthma; Dermatitis, Atopic; Disease Models, Animal; Enterotoxins; Female; Immunization; Immunoglobulin E; Inflammation; Male; Mast Cells; Mice; Mice, Inbred C57BL; Ovalbumin; Peritonitis; Staphylococcus aureus

Although natural killer T cells (NKT cells) are altered in obese asthmatic mice, their function remains completely unclear. To further explore the potential mechanism of NKT cells in airway inflammation of obesity-associated asthma, we examined the effects of α-galactosylceramide (KRN7000) on airway inflammation in obese asthmatic mice. Male C57BL/6J mice were divided into five groups: (1) control; (2) asthma; (3) A + KRN, asthma with KRN7000; (4) obese asthma; and (5) OA + KRN, obese asthma with KRN7000. Cytometric bead array (CBA) was used to detect interleukin-4 (IL-4), IL-10, tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ) in the serum. Flow cytometry was used to detect NKT cells and CD69

Topics: Adjuvants, Immunologic; Animals; Asthma; Diet, High-Fat; Galactosylceramides; Inflammation Mediators; Killer Cells, Natural; Male; Mice; Mice, Inbred C57BL; Obesity; Ovalbumin

The aim of this study is to investigate the effects of ganoderic acid A (GAA) on OVA-induced asthma in mice. Mouse asthma model was established by ovalbumin (OVA) in vitro. Diff-Quik staining was used to observe the total numbers of cells and the number of classification cells in each group, and HE staining was used to observe lung inflammation in lung tissue sections. ELISA was used to detect the effect of GAA on the levels of interleukin-4 (IL-4), IL-5, and IL-13 in serum and lung tissue. The expression levels of TLR/NF-κB were detected by Western blot. Immunohistochemistry was used to observe the expression changes of TLR4 and P-P65. Compared with the normal group, the inflammatory cell count, IL-4, IL-5, and IL-13 expression in the model group increased, and TLR/NF-kB signal protein expression increased. Compared with the model group, in GAA group, the number of inflammatory cells, the expression of IL-4, IL-5, and IL-13 decreased, and the expression of TLR/NF-kB signaling protein decreased. GAA regulated lung inflammation in asthmatic mice by inhibiting TLR/NF-kB signaling pathway.

Topics: Animals; Asthma; Cytokines; Dose-Response Relationship, Drug; Female; Heptanoic Acids; Inflammation Mediators; Lanosterol; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Toll-Like Receptor 4

Ginsenoside Rh1 (Rh1) has anti-inflammatory effects in asthma mice, but the underlying mechanism remains unclear. BALB/c mice were sensitized and challenged with ovalbumin (OVA) to construct asthma model. Mice received Rh1 or tiotropium bromide 0.5 h before OVA challenge. Airway morphology and airway remodeling were assessed by HE staining and Masson's trichrome staining, respectively. Th1/Th2 cytokines in serum or broncho alveolar lavage fluid (BALF) were measured by ELISA kits. Rh1 significantly alleviated the lung resistance and airway resistance, and reduced the number of total inflammation cells, eosinophils, neutrophils, and lymphocytes in BALF of the asthmatic mice. The morphological changes and collagen deposition of airway were also reduced by Rh1 in asthmatic mice. The increase of Eotaxin, IL-4, IL-5, IL-13, and IL-33 and the decrease of IL-12 and IFN-γ in both BALF and serum of OVA exposed mice were reversed by Rh1. Rh1 attenuates OVA-induced asthma in the mice model by regulating Th1/Th2 cytokines balance.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Ginsenosides; Interferon-gamma; Interleukins; Mice; Mice, Inbred BALB C; Ovalbumin; Th1 Cells; Th2 Cells

TMEM16A, a Ca

Topics: Animals; Anoctamin-1; Asthma; Chloride Channels; Goblet Cells; Metaplasia; Mice; Mucus; Ovalbumin; Respiratory Mucosa

Asthma, a well-known disease with high morbidity, is characterized by chronic airway inflammation. However, the allergic inflammation mechanisms of follistatin-like protein 1 (FSTL1) have not been elucidated. This study aims to investigate the effects of FSTL1 in ovalbumin (OVA)-induced mice and macrophages on nucleotide-binding domain and leucine-rich repeat protein 3 (NLRP3)/interleukin-1β (IL-1β) signaling pathway.. The present results uncovered that Fstl1. The study results showed that FSTL1 played an important role in allergic airway inflammation by activating NLRP3/IL-1β. Hence, inhibition FSTL1 could be applied as a therapeutic agent against asthma.

Topics: Aged; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Line; Cytokines; Female; Follistatin-Related Proteins; Furans; Humans; Indenes; Lung; Macrophages; Male; Mice, Inbred C57BL; Middle Aged; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Signal Transduction; Sulfonamides

Starting from our previously described PI3Kγ inhibitors, we describe the exploration of structure-activity relationships that led to the discovery of highly potent dual PI3Kγδ inhibitors. We explored changes in two positions of the molecules, including macrocyclization, but ultimately identified a simpler series with the desired potency profile that had suitable physicochemical properties for inhalation. We were able to demonstrate efficacy in a rat ovalbumin challenge model of allergic asthma and in cells derived from asthmatic patients. The optimized compound, AZD8154, has a long duration of action in the lung and low systemic exposure coupled with high selectivity against off-targets.

Topics: Animals; Asthma; Class I Phosphatidylinositol 3-Kinases; Class Ib Phosphatidylinositol 3-Kinase; Crystallography, X-Ray; Humans; Leukocytes, Mononuclear; Male; Molecular Structure; Ovalbumin; Phosphatidylinositol 3-Kinases; Protein Binding; Protein Kinase Inhibitors; Rats, Inbred BN; Structure-Activity Relationship; Sulfonamides; Thiazoles

Asthma is a chronic inflammatory disease, characterized by airway inflammation, hyperresponsiveness, and bronchial smooth muscle contraction. Qingfei Xiaoyan Wan (QFXYW), a traditional Chinese formula, has been shown to exert anti-asthma effects and immune response in multiple diseases.. In this study, we evaluated the therapeutic mechanism of QFXYW in the suppression of allergic asthma by integrating of transcriptomics and system pharmacology.. BALB/c mice were sensitized with ovalbumin (OVA) to establish the allergic asthma model, and its success was confirmed with behavioral observations. Lung histopathological analysis, inflammatory pathology scores, transcription factors were used to evaluate the effects of QFXYW on allergic asthma. The therapeutic mechanism of QFXYW in treating allergic asthma through integrated transcriptomics and system pharmacology was then determined: hub genes were screened out by topological analysis and functional enrichment analysis were performed to identify key signaling pathway. Subsequently, quantitative RP-PCR and protein array were performed to detect the mRNA of hub genes and to predict the key pathway in OVA-induced allergic asthma, respectively.. Our results demonstrated that QFXYW could significantly attenuate inflammatory cell infiltration, mucus secretion, and epithelial damage. The transcriptomics analysis found the six hub genes with the highest values- CXCL10, CXCL2, CXCL1, IL-6, CCL-5, and CCL-4 were screened out. Functional enrichment analysis showed that the differentially expressed genes (DEGs) were mainly enriched in the inflammatory response and cytokine signaling pathway. Moreover, the quantitative RT-PCR verification experiment found the CXCL2 and CXCL1 were significantly suppressed after treatment with QFXYW. The results of protein array showed that QFXYW inhibited the multi-cytokines of OVA-induced allergic asthma via cytokine signaling pathway.. QFXYW may have mediated OVA-induced allergic asthma mainly through the hub genes CXCL2, CXCL1, and the cytokine signaling pathway. This finding will offer a novel strategy to explore effective and safe mechanism of Traditional Chinese Medicine (TCM) formula to treat allergic asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Drugs, Chinese Herbal; Female; Gene Expression Regulation; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Transcriptome

Asthma is a chronic and recurring airway disease, which related to mast cell activation. Many compounds derived from Chinese herbal medicine has promising effects on stabilizing mast cells and decreasing inflammatory mediator production. Safranal, one of the active compounds from. OVA-induced asthma and PSA model were used to evaluate the effect of safranal. Safranal reduced the level of serum IgE, the number of mast cells in lung tissue were decreased and Th1/Th2 cytokine levels were normalized in OVA-induced asthma model. Furthermore, safranal inhibited BMMCs degranulation and inhibited the production of LTC. Safranal alleviates OVA-induced asthma, inhibits mast cell activation and PSA reaction. The possible mechanism occurs through the inhibition of the MAPKs and NF-κB pathways.

Topics: Allergens; Animals; Anti-Inflammatory Agents; Asthma; Cell Degranulation; Cyclohexenes; Cytokines; Disease Models, Animal; Disease Susceptibility; Female; Immunoglobulin E; Inflammation Mediators; Mast Cells; Mice; NF-kappa B; Ovalbumin; Signal Transduction; Terpenes

The phenotype and function of immature DC (DCia), DClps or IL-10-activated-DC (DC10) were determined. OVA-sensitized/challenged mice were treated with OVA-pulsed DCia or DClps or DC10. We assessed the changes of histopathology, serum total IgE level, pulmonary signal transducers and activators of transcription (STAT), pulmonary regulatory T cells (Tregs), and airway recall responses to OVA rechallenge, including proliferation and cytokine secretory function of pulmonary memory CD4. DClps exhibited low levels of CD80 and MHCII and increased levels of anti-inflammatory cytokines such as IL-10 and TGF-β. Additionally, DClps treatment dramatically diminished infiltration of inflammatory cells, eosinophilia, serum IgE and STAT6 phosphorylation level, increased the number of pulmonary Tregs. In addition, DClps treatment decreased the proliferation of pulmonary memory CD4. LPS stimulation may lead to a tolerogenic phenotype on DC, and thereby alleviated the Th2 immune response of asthmatic mice, possibly by secreting anti-inflammatory cytokines, inhibiting pulmonary memory CD4

Topics: Animals; Asthma; Biomarkers; CD4 Lymphocyte Count; Cellular Microenvironment; Cytokines; Dendritic Cells; Disease Management; Disease Models, Animal; Disease Susceptibility; Female; Gene Expression; Immunologic Memory; Immunophenotyping; Lipopolysaccharides; Mice; Ovalbumin; Phosphorylation; STAT6 Transcription Factor; T-Lymphocyte Subsets

The detailed pathogenesis of eosinophilic bronchitis (EB) remains unclear. Transglutaminase 2 (TG2) has been implicated in many respiratory diseases including asthma. Herein, we aim to assess preliminarily the relationship of TG2 with EB in the context of the development of an appropriate EB model through ovalbumin (OVA) sensitization and challenge in the C57BL/6 mouse strain. Our data lead us to propose a 50 μg dose of OVA challenge as appropriate to establish an EB model in C57BL/6 mice, whereas a challenge with a 400 μg dose of OVA significantly induced asthma. Compared to controls, TG2 is up-regulated in the airway epithelium of EB mice and EB patients. When TG2 activity was inhibited by cystamine treatment, there were no effects on airway responsiveness; in contrast, the lung pathology score and eosinophil counts in bronchoalveolar lavage fluid were significantly increased whereas the cough frequency was significantly decreased. The expression levels of interleukin (IL)-4, IL-13, IL-6, mast cell protease7 and the transient receptor potential (TRP) ankyrin 1 (TRPA1), TRP vanilloid 1 (TRPV1) were significantly decreased. These data open the possibility of an involvement of TG2 in mediating the increased cough frequency in EB through the regulation of TRPA1 and TRPV1 expression. The establishment of an EB model in C57BL/6 mice opens the way for a genetic investigation of the involvement of TG2 and other molecules in this disease using KO mice, which are often generated in the C57BL/6 genetic background.

Topics: Animals; Asthma; Bronchitis; Cystamine; Cytokines; Disease Models, Animal; Eosinophils; Gene Expression Regulation; GTP-Binding Proteins; Humans; Inflammation; Lung; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Protein Glutamine gamma Glutamyltransferase 2; Transglutaminases; TRPA1 Cation Channel

The small GTPase RhoA and its downstream effectors are critical regulators in the pathophysiological processes of asthma. The underlying mechanism, however, remains undetermined. Here, we generated an asthma mouse model with RhoA-conditional KO mice (Sftpc-cre;RhoAfl/fl) in type II alveolar epithelial cells (AT2) and demonstrated that AT2 cell-specific deletion of RhoA leads to exacerbation of allergen-induced airway hyperresponsiveness and airway inflammation with elevated Th2 cytokines in bronchoalveolar lavage fluid (BALF). Notably, Sftpc-cre;RhoAfl/fl mice showed a significant reduction in Tgf-β1 levels in BALF and lung tissues, and administration of recombinant Tgf-β1 to the mice rescued Tgf-β1 and alleviated the increased allergic airway inflammation observed in Sftpc-cre;RhoAfl/fl mice. Using RNA sequencing technology, we identified Slc26a4 (pendrin), a transmembrane anion exchange, as the most upregulated gene in RhoA-deficient AT2 cells. The upregulation of SLC26A4 was further confirmed in AT2 cells of asthmatic patients and mouse models and in human airway epithelial cells expressing dominant-negative RHOA (RHOA-N19). SLA26A4 was also elevated in serum from asthmatic patients and negatively associated with the percentage of forced expiratory volume in 1 second (FEV1%). Furthermore, SLC26A4 inhibition promoted epithelial TGF-β1 release and attenuated allergic airway inflammation. Our study reveals a RhoA/SLC26A4 axis in AT2 cells that functions as a protective mechanism against allergic airway inflammation.

Topics: Alveolar Epithelial Cells; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Humans; Lung; Mice; Ovalbumin; Recombinant Proteins; rhoA GTP-Binding Protein; Sulfate Transporters; Symptom Flare Up; Transforming Growth Factor beta1

Asthma is one of the most common respiratory diseases. Lack of response or poor adherence to corticosteroids demands the development of new drug candidates for asthma. Endogenous nucleosides could be potential options since uridine has been reported to have an anti-inflammatory effect in asthma model. However, its molecular pathways and whether other nucleosides have similar therapeutic effects remain untouched. Thus, we herein report our investigation into the anti-inflammatory effects of guanosine and uridine, and the related inner signaling pathways in asthma model. Present study shows that administration of guanosine or uridine can reduce lung inflammation in OVA-challenged mice. Total cell counts in BALF, cytokines such as IL-4, IL-6, IL-13, OVA-specific IgE and mRNA level of Cxcl1, Cxlc3, IL-17 and Muc5ac were decreased in asthmatic mice after treatment. Besides, the production of IL-6 in LPS/IFN-γ induced THP-1 cells was also decreased by both nucleosides. In vivo and in vitro expressions of key molecules in the MAPK and NF-κB pathways were reduced after the treatment of both compounds. These findings suggest that guanosine has a similar potential therapeutic value in asthma as uridine and they exert anti-inflammatory effects through suppression of the MAPK and NF-κB pathways.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Guanosine; Inflammation; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Uridine

Topics: Animals; Asthma; Female; Janus Kinases; Mice; Mice, Inbred C57BL; Ovalbumin; Pulmonary Surfactant-Associated Protein A; Signal Transduction; STAT Transcription Factors; Th1 Cells; Th17 Cells

Epidemiologic evidence suggests that PM2.5 exposure aggravates asthma, but the molecular mechanisms are not fully discovered.. Ovalbumin (OVA)-induced mice exposed to PM2.5 were constructed. Pathological staining and immunofluorescence were performed in in vivo study. Gene set enrichment analysis (GSEA) was performed to identify the pathway involved in asthma severity by using U-BIOPRED data (human bronchial biopsies) and RNA-seq data (Beas-2B cells treated with PM2.5). Lentiviruses transfection, Real-time qPCR, immunofluorescence staining and trans-epithelial electrical resistance (TEER) measurement were performed for mechanism exploration in vitro.. PM2.5 exposure aggravated airway inflammation and mucus secretion in OVA-induced mice. Based on transcriptome analysis of mild-to-severe asthma from human bronchial biopsies, gene set enrichment analysis (GSEA) showed that up-regulated reactive oxygen species (ROS) pathway gene set and down-regulated apical junction gene set correlated with asthma severity. Consistent with the analysis of mild-to-severe asthma, after PM2.5 exposure, the ROS pathway in Beas-2B cells was up-regulated with the down-regulation of apical junction. The expression levels of genes involved in the specific gene sets were validated by using qPCR. The mRNA levels of junction genes, ZO-1, E-cadherin and Occludin, were significantly decreased in cells exposed to PM2.5. Moreover, it confirmed that inhibition of ROS recovered the expression levels of E-cadherin, Occludin and ZO-1, and ameliorated inflammation and mucus secretion in airway in OVA-induced mice exposed to PM2.5. Meanwhile, ROS level was elevated by PM2.5. By checking trans-epithelial electrical resistance (TEER) value, we also found that epithelial barrier was damaged after PM2.5 exposure. Importantly, Stanniocalcin 2 (STC2) was identified as a key gene in regulation of epithelial barrier. It showed that STC2 expression was up-regulated by PM2.5, which was recovered by NAC as well. Over-expression of STC2 could decrease the expression levels of ZO-1, Occludin and E-cadherin. Contrarily, suppression of STC2 could increase the expression levels of ZO-1, Occludin and E-cadherin reduced by PM2.5.. By using transcriptome analysis, we revealed that STC2 played a key role in PM2.5 aggravated airway dysfunction through regulation of epithelial barrier in OVA-induced mice.

Topics: Animals; Asthma; Disease Models, Animal; Gene Expression Profiling; Glycoproteins; Humans; Inflammation; Intercellular Signaling Peptides and Proteins; Mice; Ovalbumin; Particulate Matter; Reactive Oxygen Species; Respiratory Mucosa; Transcriptome; Up-Regulation

Bronchial asthma (BA) is a heterogeneous allergic respiratory disease with diverse inflammatory symptoms, pathology, and responses to treatment. Thyme is a natural product which is consisted of multiple phenolic compounds of therapeutic significance for treatment of cough and bronchitis. This study evaluated the efficacy of thyme oil against ovalbumin (OVA)-induced BA in an experimental rabbit model. Forty male rabbits were divided into four equal groups [control group (G1), OVA (G2), thyme oil (G3), and OVA plus thyme oil (G4)]. Animals were treated for 30 days, and clinical, histopathological (HP), histochemical (HC), immunohistochemical (IHC), morphometric, biochemical and flow cytometry methods were performed, followed by statistical analysis. All used methods revealed normal structure of the lung tissues in rabbits of G1 and G3. In contrast, the clinical examination of G2 rabbits revealed an obvious increase in the respiratory rate, sneezing and wheezing, whereas the HP, HC and IHC techniques exhibited substantial inflammatory changes in the peribronchio-vascular lung tissues with thinning, degeneration, apoptosis (using the TUNEL assay), necrosis, and shedding of the airway epithelium. Furthermore, the morphometric results confirmed significant increases in the numbers of inflammatory cells, goblet cells, eosinophils and apoptotic cells from (12, 0, 2, 2 cells) to (34,10, 16, 18 cells) respectively, as well as the area percentage of collagen fiber deposition and immunoexpression of eotaxin-1/10 high power fields. Additionally, the biochemical results revealed significant increases in the serum levels of TSLP, IL-4, IL-5, IL-9, IL-13, IgE and eotaxin-1 cytokines from (140, 40, 15, 38, 120, 100, 48) pg./ml to (360, 270, 130, 85, 365, 398, 110) pg./ml respectively, while analysis of ROS by flow cytometry revealed remarkable oxidative stress effects in G2 rabbits. On the other hand, treatment of rabbits with thyme oil in G4 substantially alleviated all OVA-induced alterations. Overall, our findings indicate for the first time that thyme oil can ameliorate OVA-induced BA via its immunomodulatory, anti-inflammatory, antiapoptotic, and antioxidant effects on the lung tissues of rabbits.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antioxidants; Asthma; Cytokines; Goblet Cells; Immunoglobulin E; Lung; Male; Ovalbumin; Plant Oils; Rabbits; Reactive Oxygen Species; Th2 Cells; Thymus Plant

Asthma is a frequently occurring respiratory disease with an increasing incidence around the world. Airway inflammation and remodeling are important contributors to the occurrence of asthma. We conducted this study aiming at exploring the effect of Histone deacetylase 4 (HDAC4)-mediated Kruppel-like factor 5 (KLF5)/Slug/CXC chemokine ligand-12 (CXCL12) axis on the development of asthma in regulation of airway inflammation and remodeling.. An asthmatic rat model was induced by ovalbumin (OVA) irrigation, and determined HDAC4, KLF5, Slug, and CXCL12 expression in the lung tissues by RT-qPCR and Western blot assay. OVA was also used to induce a cell model of asthma in human BEAS-2B and HBE135-E6E7bronchial epithelial cells. The airway hyperresponsiveness (AHR), and expression of inflammatory cytokines in model mice were examined using methacholine challenge test and ELISA. The biological behaviors were measured in asthma model bronchial smooth muscle cells (BSMCs) following loss- and gain- function approaches. The interactions between HDAC4, KLF5, Slug, and CXCL12 were also detected by IP assay, dual luciferase gene reporter assay, and ChIP.. HDAC4 was upregulated in lung tissues of OVA-induced asthmatic mice, and inhibition of HDAC4 alleviated the airway inflammation and remodeling. HDAC4 increased KLF5 transcriptional activity through deacetylation; deacetylated KLF5 bound to the promoter of Slug and transcriptionally upregulated Slug expression, which in turn increased the expression of CXCL12 to promote the inflammation in bronchial epithelial cells and thus induce the proliferation and migration of BSMCs.. Collectively, HDAC4 deacetylates KLF5 to upregulate Slug and CXCL12, thereby causing airway remodeling and facilitating progression of asthma.

Topics: Airway Remodeling; Animals; Asthma; Chemokines, CXC; Disease Models, Animal; Histone Deacetylases; Kruppel-Like Transcription Factors; Ligands; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Rats; Repressor Proteins

Over the past few years, ozone has been identified as a potential risk factor for exacerbating asthma. However, few attempts have been made to prevent the progression of ozone-exacerbated asthma. This study investigated the attenuating effects of melatonin on ozone-aggravated allergic asthma, and explored the changes to the transient receptor potential vanilloid 1 (TRPV1)-nuclear factor erythroid-derived 2-related factor 2 (Nrf2) pathway associated with melatonin treatment. The levels of TRPV1 and calcitonin gene-related peptides (CGRP) in lung tissue were detected by immunohistochemistry, western blot, and enzyme-linked immunosorbent assay (ELISA). The Nrf2 signaling involved proteins and mRNA were evaluated by western blot and RT-qPCR. The change of Immunoglobulin E (IgE) and T helper (Th) 2 and Th17 cytokines in serum and bronchoalveolar lavage fluid (BALF) was determined by ELISA. Recruitment of inflammatory cells in BALF, histopathological changes, and airway hyperresponsiveness (AHR) were also determined in lung tissues. Our results indicated that melatonin treatment significantly reduced oxidative stress, as indicated by levels of glutathione (GSH), malonaldehyde (MDA), and 8-hydroxy-2-deoxyguanosine (8-OH-dG). Moreover, ozone-exacerbated asthma symptoms, such as inflammatory cell infiltration, levels of serum immunoglobulin, Th2 and Th17 cytokines in BALF, obvious changes in lung histology, and AHR, were all ameliorated by melatonin treatment. Interestingly, melatonin not only markedly decreased the protein levels of TRPV1 and CGRP, but also enhanced the expression of Nrf2, quinone oxidoreductase-1 (NQO-1), and heme oxygenase-1 (HO-1). Taken together, our results demonstrate that melatonin administration could antagonize ozone-exacerbated asthma by inhibiting the TRPV1 channel and stabilizing the Nrf2 pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Lung; Melatonin; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; Ovalbumin; Ozone; Transient Receptor Potential Channels; TRPV Cation Channels

Lysosome-associated membrane glycoprotein 3 (LAMP3) is a type I transmembrane protein of the LAMP protein family with a cell-type-specific expression in alveolar type II cells in mice and hitherto unknown function. In type II pneumocytes, LAMP3 is localized in lamellar bodies, secretory organelles releasing pulmonary surfactant into the extracellular space to lower surface tension at the air/liquid interface. The physiological function of LAMP3, however, remains enigmatic. We generated Lamp3 knockout mice by CRISPR/Cas9. LAMP3 deficient mice are viable with an average life span and display regular lung function under basal conditions. The levels of a major hydrophobic protein component of pulmonary surfactant, SP-C, are strongly increased in the lung of Lamp3 knockout mice, and the lipid composition of the bronchoalveolar lavage shows mild but significant changes, resulting in alterations in surfactant functionality. In ovalbumin-induced experimental allergic asthma, the changes in lipid composition are aggravated, and LAMP3-deficient mice exert an increased airway resistance. Our data suggest a critical role of LAMP3 in the regulation of pulmonary surfactant homeostasis and normal lung function.

Topics: Airway Resistance; Alveolar Epithelial Cells; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Gene Editing; Gene Expression Regulation; Homeostasis; Lipidomics; Lung; Lysosomal-Associated Membrane Protein 3; Mice; Mice, Knockout; Ovalbumin; Protein Isoforms; Pulmonary Alveoli; Pulmonary Surfactant-Associated Protein C; Pulmonary Surfactants; Respiratory Function Tests; Signal Transduction

Curcumol exhibits anti-inflammatory effect, but its effect on chronic asthma lacked research. Therefore, this study explored the role of curcumol in asthma.. A chronic asthmatic mice model was established by ovalbumin induction. After treatment with curcumol, airway resistance in mice was detected by forced oscillation technique. The histopathological features of airway tissues, pulmonary inflammation, and inflammation cell recruitment in the bronchoalveolar lavage fluid (BALF) of mice were detected by hematoxylin-eosin staining. Collagen deposition in the airways of mice was examined by Masson staining. The secretion of ovalbumin-IgE, IL-4, IL-5, IL-13 in mouse serum and VEGFA secretion in BALF were analyzed by ELISA. Finally, the expressions of β-catenin, Wnt5a, VEGFA, TGF-β1, Fibronectin, and MMP-9 in mice lung tissues were determined by Western blot or immunohistochemical.. Curcumol attenuated airway hyperresponsiveness, airway remodeling, and pulmonary inflammation in chronic asthmatic mice. Curcumol relieved collagen deposition in airway tissues, inflammation cell recruitment in BALF, and reduced the up-regulation of serum ovalbumin-IgE, IL-4, IL-5, and IL-13 and BALF VEGFA in chronic asthmatic mice. In addition, curcumol attenuated the up-regulated expressions of β-catenin, Wnt5a, VEGFA, TGF-β1, Fibronectin, and MMP-9 in the lung tissues of chronic asthmatic mice, but curcumol treatment did not show such effects on healthy mice.. Our findings revealed that curcumol could ameliorate lung inflammation and airway remodeling by inhibiting the abnormal activation of the Wnt/β-catenin pathway in chronic asthmatic mice, indicating that curcumol could be used as a novel anti-asthma drug for basic and clinical research.

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Sesquiterpenes; Wnt Signaling Pathway

Topics: Allergens; Animals; Anti-Allergic Agents; Asthma; Dendritic Cells; Disease Models, Animal; Gastrointestinal Microbiome; Humans; Hypersensitivity; Intestinal Mucosa; Lactobacillus; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; T-Lymphocytes, Regulatory

Asthma oxidative stress disturbances seem to enable supplementary proinflammatory pathways, thus contributing to disease development and severity. The current study analyzed the impact of two types of oral vitamin D (VD) supplementation regimens on the redox balance using a murine model of acute ovalbumin-induced (OVA-induced) asthmatic inflammation. The experimental prevention group received a long-term daily dose of 50 µg/kg (total dose of 1300 µg/kg), whereas the rescue group underwent a short-term daily dose of 100 µg/kg (total dose of 400 µg/kg). The following oxidative stress parameters were analyzed in serum, bronchoalveolar lavage fluid (BALF) and lung tissue homogenate (LTH): total oxidative status, total antioxidant response, oxidative stress index, malondialdehyde and total thiols. Results showed that VD significantly reduced oxidative forces and increased the antioxidant capacity in the serum and LTH of treated mice. There was no statistically significant difference between the two types of VD supplementation. VD also exhibited an anti-inflammatory effect in all treated mice, reducing nitric oxide formation in serum and the expression of nuclear factor kappa B p65 in the lung. In conclusion, VD supplementation seems to exhibit a protective role in oxidative stress processes related to OVA-induced acute airway inflammation.

Topics: Animals; Asthma; Disease Models, Animal; Female; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Vitamin D

Appropriate drug treatment for smoking asthmatics is uncertain because most smokers with asthma are less sensitive to treatment with glucocorticoids compared with non-smokers with asthma. We hypothesized that roflumilast (Rof), a selective phosphodiesterases-4 inhibitor regarded as an add-on therapy for chronic obstructive pulmonary disease, might be more effective than glucocorticoids for improving asthma in smokers. To investigate this hypothesis, we compared the therapeutic effects of dexamethasone (Dex) and Rof in a mouse model of ovalbumin-induced asthma with or without concurrent cigarette smoke (CS) exposure for 2 weeks. We found that recurrent asthma attacks increased lung tissue resistance. CS exposure in asthmatic mice decreased the central airway resistance, increased lung compliance, and attenuated airway hyper-responsiveness (AHR). CS exposure in asthmatic mice also increased the number of neutrophils and macrophages in the bronchoalveolar fluid. Treatment with Dex in asthmatic mice without CS exposure reduced airway resistance, AHR and airway eosinophilia. In asthmatic mice with CS exposure, however, Dex treatment unexpectedly increased lung tissue resistance and restored AHR that had been otherwise suppressed. Dex treatment in asthmatic mice with CS exposure inhibited eosinophilic inflammation but conversely exacerbated neutrophilic inflammation. On the other hand, treatment with Rof in asthmatic mice without CS exposure reduced airway resistance and airway eosinophilia, although the inhibitory effect of Rof on AHR was unremarkable. In asthmatic mice with CS exposure, Rof treatment did not exacerbate lung tissue resistance but modestly restored AHR, without any significant effects on airway inflammation. These results suggest that CS exposure mitigates sensitivity to both Dex and Rof. In asthmatic mice with CS exposure, Dex is still effective in reducing eosinophilic inflammation but increases lung tissue resistance, AHR and neutrophilic inflammation. Rof is ineffective in improving lung function and inflammation in asthmatic mice with CS exposure. This study did not support our initial hypothesis that Rof might be more effective than glucocorticoids for improving asthma in smokers. However, glucocorticoids may have a detrimental effect on smoking asthmatics.

Topics: Aminopyridines; Animals; Asthma; Benzamides; Bronchoalveolar Lavage Fluid; Cyclopropanes; Dexamethasone; Disease Models, Animal; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Smoking

Asthma is a complex inflammatory disorder that plagues a large number of people. Schisandrin B is an active ingredient of the traditional Chinese herbal medicine Schisandra with various proven physiological activities such as anti-inflammatory and antioxidant activities. In this study, we explored the anti-inflammatory and antioxidant effects and provided the mechanistic insights into the activity of schisandrin B in a mouse model of ovalbumin- (OVA-) induced allergic asthma.. Male BALB/c mice were sensitized and challenged with OVA to induce asthma and treated with various doses (15 mg/kg, 30 mg/kg, and 60 mg/kg) of SCH to alleviate the features of allergic asthma, airway hyperresponsiveness, inflammatory response, OVA-specific immunoglobulin (Ig)E level, and pathological injury.. Schisandrin B significantly attenuated the airway hyperresponsiveness induced by OVA. Moreover, schisandrin B administration suppressed inflammatory responses, reduced the level of IgE, and attenuated pathological injury. Mechanistically, schisandrin B treatment promoted the activation of nuclear erythroid 2-related factor 2 (Nrf2), but suppressed the stimulation of the NF-. Taken together, our study suggests that schisandrin B attenuates the features of asthmatic lungs by inhibiting the NF-

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cyclooctanes; Disease Models, Animal; Humans; Lignans; Lung; Male; Mice; NF-E2-Related Factor 2; NF-kappa B; Ovalbumin; Oxidative Stress; Polycyclic Compounds; Signal Transduction; Specific Pathogen-Free Organisms

Respiratory viral infections are one of the leading causes of need for emergency care and hospitalizations in asthmatic individuals, and airway-secreted cytokines are released within hours of viral infection to initiate these exacerbations. IL-33, specifically, contributes to these allergic exacerbations by amplifying type 2 inflammation. We hypothesized that blocking IL-33 in RSV-induced exacerbation would significantly reduce allergic inflammation.. Sensitized BALB/c mice were challenged with aerosolized ovalbumin (OVA) to establish allergic inflammation, followed by RSV-A2 infection to yield four treatment groups: saline only (Saline), RSV-infected alone (RSV), OVA alone (OVA), and OVA-treated with RSV infection (OVA-RSV). Lung outcomes included lung mRNA and protein markers of allergic inflammation, histology for mucus cell metaplasia and lung immune cell influx by cytospin and flow cytometry.. While thymic stromal lymphopoietin (TSLP) and IL-33 were detected 6 h after RSV infection in the OVA-RSV mice, IL-23 protein was uniquely upregulated in RSV-infected mice alone. OVA-RSV animals varied from RSV- or OVA-treated mice as they had increased lung eosinophils, neutrophils, group 2 innate lymphoid cells (ILC2) and group 3 innate lymphoid cells (ILC3) detectable as early as 6 h after RSV infection. Neutralized IL-33 significantly reduced ILC2 and eosinophils, and the prototypical allergic proteins, IL-5, IL-13, CCL17 and CCL22 in OVA-RSV mice. Numbers of neutrophils and ILC3 were also reduced with anti-IL-33 treatment in both RSV and OVA-RSV treated animals as well.. Taken together, our findings indicate a broad reduction in allergic-proinflammatory events mediated by IL-33 neutralization in RSV-induced asthma exacerbation.

Topics: Animals; Asthma; Female; Interleukin-33; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses

Asthma is an inflammatory disease that affects many people around the world, especially persons at paediatric age group. The effectiveness of tyrosol, a natural phenolic compound, was examined in the asthma model induced by ovalbumin (OVA). For this purpose, four groups, each consisting of eight rats, were arranged. For 21 days, physiological saline solution was treated to the control group and OVA was treated to the groups of OVA, OVA + dexamethasone (Dexa) and OVA + tyrosol groups, intraperitoneally and through inhalation. Additionally, 0.25 mg/kg Dexa was treated to the OVA + Dexa group and 20 mg/kg tyrosol to the OVA + tyrosol group by oral gavage. Serum, blood, bronchoalveolar lavage fluid (BALF) and lung tissues of the rats were examined. It was observed that MDA level decreased, GSH level and GPx activity increased, and there was no change in CAT activity in lung tissues of the tyrosol treatment groups. It was also observed that NF-κB, TNF-α, IL-4, IL-5, IL-13, IFN-γ and IgE levels decreased compared to the OVA group in lung tissue and serum samples except for serum NF-κB and IL-4. However, no effect on IL-1 β level was observed. In addition, it was determined that tyrosol treatment increased the IL-10 level on both tissue samples. The results of the histopathological investigation of lung tissue showed that tyrosol significantly ameliorated OVA-induced histopathological lesions. Additionally, PAS staining showed that mucus hypersecretion was significantly reduced with the use of tyrosol. In addition, it was determined that the number of eosinophils decreased significantly in blood and BALF samples. The obtained results showed that tyrosol possessed antioxidant and anti-inflammatory features on OVA-induced rats and preserved tissue architecture.

Topics: Allergens; Animals; Anti-Inflammatory Agents; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Catalase; Cytokines; Disease Models, Animal; Eosinophils; Glutathione; Glutathione Peroxidase; Immunoglobulin E; Leukocyte Count; Lung; NF-kappa B; Ovalbumin; Phenylethyl Alcohol; Rats, Wistar

Topics: Allergens; Animals; Asthma; Cells, Cultured; Dexamethasone; Disease Models, Animal; Drug Resistance; Forkhead Transcription Factors; Humans; Interleukin-27; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Protein Isoforms; Receptors, Glucocorticoid; Respiratory Hypersensitivity; T-Lymphocytes, Regulatory

Lung interstitial macrophages (IMs) can be polarized towards an alternative activation phenotype in ovalbumin (OVA)-induced asthmatic mice. However, the role of alternative activation of lung IMs in Th2 cell responses in the asthmatic murine is still unclear. Here, we leverage an anti-F4/80 treatment which has been shown to selectively deplete IMs in mice and investigate how this treatment modulates Th2 cell responses in lung and whether the modulation is dependent on lung IMs in murine models of asthma. We show that anti-F4/80 treatment alleviates Th2 cell responses in mice immunized and challenged with OVA or house dust mite (HDM). The anti-F4/80 treatment does not target lung alveolar macrophages (AMs) in OVA-induced asthmatic mice or impact the abundance of other immune cell types, including B cells, T cells, and NK cells in wild-type mice. However, this treatment does inhibit the expression of polarized markers of alternatively activated macrophages, including arginase-1, Ym-1, and Fizz-1 in the lung tissues from OVA-induced asthmatic mice. Furthermore, we find that the inhibitory effects of anti-F4/80 treatment on Th2 cell responses can be reversed upon adoptive transfer of lung IMs. Taken together, our data show that anti-F4/80 treatment attenuates Th2 cell responses, which is at least partially related to depletion of lung IMs in murine models of asthma. This suggests that targeted lung IMs may provide a potential therapeutic protocol for the treatment of asthmatics.

Topics: Allergens; Animals; Asthma; Cytokines; Female; Inflammation; Lung; Macrophages; Mice; Mice, Inbred BALB C; Ovalbumin; Pyroglyphidae; Th2 Cells

Angiogenesis is a key characteristic of asthma airway remodeling. By releasing cationic granule proteins, such as major basic protein (MBP), activated eosinophils play a prominent role in asthma, but the underlying mechanisms are still not fully understood. In this study, we demonstrated that fibroblast growth factor-binding protein 1 (FGFBP1) was dramatically upregulated in airway epithelial cell lines treated by poly-L-arginine (PLA), a mimic of MBP. Elevated FGFBP1 expression was also detected in asthma clinical samples, as well as in ovalbumin (OVA)-induced chronic asthma mouse models. PLA enhanced FGFBP1 expression through activation of the mechanistic target of rapamycin complex 1-signal transducer and activator of transcription 3 (mTORC1-STAT3) signaling pathway. STAT3 transactivated FGFBP1 by directly binding to the promoter of the FGFBP1 gene. Furthermore, we identified that FGFBP1 secreted by PLA-treated airway epithelial cells served as a proangiogenesis factor. Lastly, we found the mTORC1-STAT3-FGFBP1 signaling pathway was activated in an OVA-induced chronic asthma model with airway remodeling features. Rapamycin treatment alleviated respiratory symptoms and reduced angiogenesis in asthmatic mice. Therefore, activation of the mTORC1-STAT3-FGFBP1 pathway in the airway epithelium contributes to the progress of angiogenesis and should be targeted for the treatment of asthma.

Topics: Animals; Asthma; Cell Count; Cell Line, Tumor; Chickens; Eosinophils; Epithelial Cells; Gene Expression Profiling; Gene Expression Regulation; Human Umbilical Vein Endothelial Cells; Humans; Intercellular Signaling Peptides and Proteins; Lung; Mechanistic Target of Rapamycin Complex 1; Mice; Models, Biological; Neovascularization, Pathologic; Ovalbumin; Peptides; Signal Transduction; STAT3 Transcription Factor; Transcription, Genetic; Up-Regulation

Eosinophilic asthma is a form of bronchial asthma that is caused by the pulmonary infiltration of eosinophils and accounts for approximately half of the patients with severe asthma. Several cell types of the immune system in synergy with the epithelial cells of the lung provoke an inflammatory response in patients with asthma. Recently, the effect of fasting on immune cells and inflammation has attracted considerable attention. Therefore, we examined whether fasting may serve as novel preventive strategy in patients with asthma. In our study, we employed a previously established mouse model of eosinophilic asthma. C57BL/6 mice were inoculated intranasally with interleukin-33 and ovalbumin (OVA) in order to induce eosinophil infiltration in the lung and subjected to a 48-h long fasting period directly after or 7 days postinoculation. We used flow cytometry to characterise infiltrated immune cells in the lung and measured the quantity of inflammatory cytokines as well as antigen-specific immunoglobins (Ig) by ELISA. Our results indicated that fasting lowered the number of eosinophilic pulmonary infiltrates in the eosinophilic asthma model mice. Furthermore, fasting suppressed anti-OVA IgG1 production. Fasting suppressed Th2 cytokine production by impairing Th2 accumulation in the lung. The findings suggest that fasting may be a novel preventive strategy for eosinophilic asthma.

Topics: Allergens; Animals; Asthma; Biomarkers; Cytokines; Disease Models, Animal; Disease Susceptibility; Eosinophils; Fasting; Immunoglobulin E; Immunoglobulin G; Immunomodulation; Lung; Mice; Ovalbumin; Th2 Cells

The calcium-binding protein S100A4 demonstrates important regulatory roles in many biological processes including tumorigenesis and inflammatory disorders such as allergy. However, the specific mechanism of the contribution of S100A4 to allergic diseases awaits further clarification.. To address the effect of S100A4 on the regulation of mast cell activation and its impact on allergy.. Bone marrow-derived cultured mast cells (BMMCs) were derived from wild-type (WT) or S100A4. Following OVA/alum-based sensitization and provocation, S100A4. S100A4 is required for mast cell functional activation, and S100A4 may participate in the regulation of allergic responses at least partly through regulating the activation of mast cells.

Topics: Animals; Asthma; Cell Degranulation; Cells, Cultured; Cytokines; Disease Models, Animal; Immunity, Humoral; Immunoglobulin E; Immunoglobulin G; Immunologic Memory; Lung; Male; Mast Cells; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Passive Cutaneous Anaphylaxis; S100 Calcium-Binding Protein A4; T-Lymphocytes

Isoalantolactone (IAL), a sesquiterpene lactone, has anti‑inflammatory activity in lipopolysaccharide (LPS)‑induced sepsis. However, it remains to be elucidated whether IAL influences asthmatic inflammation. The present study found that IAL inhibited ovalbumin (OVA)‑induced asthmatic inflammation and attenuated OVA‑induced eosinophil infiltration, immunoglobulin E generation and the production of interleukin (IL)‑4, IL‑5, C‑C motif chemokine ligand (CCL)17 and CCL22. In addition, IAL treatment with IL‑4 reduced the expression of arginase‑1, Ym‑1, CCL17 and CCL22 in bone marrow‑derived macrophages

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Chemokine CCL17; Cytokines; Female; Humans; Immunoglobulin E; Inflammation; Interleukin-4; Kruppel-Like Factor 4; Lipopolysaccharides; Lung; Macrophages; Mice; Mice, Inbred BALB C; Ovalbumin; PPAR gamma; Sesquiterpenes; STAT6 Transcription Factor

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Interleukin-6; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Progranulins

Allergic asthma and atherosclerosis are inflammatory diseases characterized by similar sets of circulating inflammatory cells, in addition to mast cells in the airway and vessel wall. Animal models and human studies provide evidence of a potential interaction between the two apparently unrelated diseases. The main objective of this study was to determine whether experimental allergic asthma is accompanied by inflammatory responses, measured as the activation of the vasculature and the presence of immune cells in the perivascular adipose tissue. For this purpose, male Dunkin Hartley guinea pigs weighing 250 - 300 g were sensitized twice with 10 μg ovalbumin dissolved in aluminium hydroxide (Al(OH)

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Guinea Pigs; Inflammation; Male; Ovalbumin

Despite an ongoing focus on the role of diet in health and disease, we have only a limited understanding of these concepts at the cellular and molecular levels. While obesity has been clearly recognized as contributing to metabolic syndrome and the pathogenesis of adult asthma, recent evidence has linked high sugar intake alone to an increased risk of developing asthma in childhood. In this study, we examined the impact of diet in a mouse model of allergic airways inflammation with a specific focus on eosinophils. As anticipated, male C57BL/6 mice gained weight on a high-calorie, high-fat diet. However, mice also gained weight on an isocaloric high-sucrose diet. Elevated levels of leptin were detected in the serum and airways of mice maintained on the high-fat, but not the high-sucrose diets. We found that diet alone had no impact on eosinophil numbers in the airways at baseline or their recruitment in response to allergen (Alternaria alternata) challenge in either wild-type or leptin-deficient ob/ob mice. However, both bronchoalveolar lavage fluid and eosinophils isolated from lung tissue of allergen-challenged mice exhibited profound diet-dependent differences in cytokine content. Similarly, while all wild-type mice responded to allergen challenge with significant increases in methacholine-dependent total airway resistance (Rrs), airway resistance in mice maintained on the isocaloric high-sucrose (but not the high-calorie/high-fat) diet significantly exceeded that of mice maintained on the basic diet. In summary, our findings revealed that mice maintained on an isocaloric high-sucrose diet responded to allergen challenge with significant changes in both BAL and eosinophil cytokine content together with significant increases in Rrs. These results provide a model for further exploration of the unique risks associated with a high-sugar diet and its impact on allergen-associated respiratory dysfunction.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Diet, High-Fat; Eosinophils; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Pneumonia; Sucrose; Sweetening Agents

Tris(1,3-dichloro-2-propyl)phosphate (TDCIPP) is an organophosphorus flame retardant that is an alternative to brominated flame retardants. Although TDCIPP can adversely affect human health, information about its effects on immune and allergic responses is scarce. We aimed to investigate the effects of dietary exposure to TDCIPP using less than the human tolerable daily intake (TDI) in allergic asthmatic mice.. Male C3H/HeJSlc mice were fed a chow diet containing TDCIPP equivalent to 0.02 μg/kg/day (low; L), 0.2 μg/kg/day (medium; M), or 2 μg/kg/day (high; H) and were intratracheally administered ovalbumin (OVA, 1 μg/animal) every 2 weeks from 5 to 11 weeks of age.. In OVA-treated mice, TDCIPP-H exposure tended to enhance pulmonary inflammation compared with vehicle exposure. TDCIPP dose-dependently decreased mRNA level of G protein-coupled estrogen receptor (GPER) in the lungs with or without OVA. OVA + TDCIPP-H treatment tended to increase the total cell number and promoted CD4. These results suggested that dietary exposure to TDCIPP at TDI level slightly enhances allergic diseases, such as allergic asthma,

Topics: Animals; Asthma; Cells, Cultured; Dietary Exposure; Flame Retardants; Lung; Male; Mice; Mice, Inbred C3H; Organophosphorus Compounds; Ovalbumin

Hanchuan Zupa Granule (HCZP) is a classic prescription of Uyghur medicine, that is used for cough and abnormal mucinous asthma caused by a cold and "Nai-Zi-Lai".. This study aimed to explore the possible molecular mechanism of HCZP in the treatment of asthma, using a network pharmacology method and in vivo experiments.. First, we conducted qualitative analysis of the chemical composition of HCZP as a basis for network pharmacology analysis. Using network pharmacology tools, the possible signaling pathways of HCZP in the treatment of asthma were obtained. An OVA-sensitized asthma model was established, and HCZP was continuously administered for one week. BALF was collected for cell counting, and serum and lung tissues were collected to analyze the expression of IgE, IL-4, IL-5, IL-13 and IFN-γ. Hematoxylin & eosin (H&E) staining was performed to assess the pathological changes in the lung tissues. Related protein expression in the lung tissues was analyzed by Western blotting for molecular mechanism exploration.. Fifty-six chemical compounds were identified by UPLC Q-TOF MS. According to the network pharmacology results, 18 active compounds were identified among the 56 compounds, and 68 target genes of HCZP in the treatment of asthma were obtained. A total of 19 pathways were responsible for asthma (P < 0.05) according to KEGG pathway analysis. In vivo results showed that OVA sensitivity induced increased respiratory system resistance and inflammatory responses, which included inflammatory cell infiltration and high levels of IgE, IL-4, IL-5 and IL-13 in serum and lung tissues. Furthermore, OVA upregulated p-PI3K, p-JNK and p-p38 expression in lung tissues. Moreover, HCZP treatment significantly downregulated respiratory system resistance, and the expression of IL-4, IL-5, IL-13 and IgE, as well as significantly improved inflammatory cell infiltration in lung tissues. Moreover, the protein expression of p-PI3K, p-JNK and p-p38 in lung tissues decreased after HCZP treatment.. HCZP significantly inhibited the OVA-induced inflammatory response via the PI3K-Akt and Fc epsilon RI signaling pathways.

Topics: Animals; Asian People; Asthma; Databases, Factual; Dexamethasone; Female; Humans; Medicine, Traditional; Mice; Mice, Inbred BALB C; Ovalbumin; Random Allocation; Reproducibility of Results

Studies on the bronchial vascular bed have revealed that the number of blood vessels in the lamina propria and under the mucosa of the lung tissue increases in patients suffering from mild to severe asthma. Thus, in this study, a new strategy was employed in respiratory system disorders by angiogenesis inhibition in an ovalbumin (OVA)-induced rat model of asthma. Twenty-one male Wistar albino rats, 8 weeks old, were randomly divided into three groups (n = 7 in each group), including (1) control group, (2) OVA-treated group, and (3) OVA + Bmab (bevacizumab drug). On days 1 and 8, 1 mg of OVA and aluminum hydroxide in sterile phosphate-buffered saline (PBS) were intraperitoneally injected to rats in groups 2 and 3. The control group was only subject to intraperitoneal injection of saline on days 1 and 8. One week after the last injection, the rats (groups 2 and 3) were exposed to OVA inhalation for 30 min at 2-day intervals from days 15 to 25. After sensitization and challenge with OVA, the OVA + Bmab group (group 3) were treated with a 5 mg/kg bevacizumab drug. Genes and protein expression of IL-1β and TNF-α and the expression of vascular endothelial growth factor (VEGF) protein were assessed by real-time PCR and immunohistochemistry respectively, in lung tissue. OVA exposure increased mucosal secretion and inflammatory cell populations in lung tissue and OVA-specific IgE level in serum. Also, VEGF and cytokine factor expression were significantly elevated in the OVA-induced asthma model (p ≤ 0.05). However, rats in OVA + Bmab group showed significantly a decrease in VEGF and IL-1β and TNF-α genes as well as proteins (p ≤ 0.05). The results showed that bevacizumab efficiently diminished bronchial inflammation via downregulation of VEGF expression, followed by inflammatory cells population and cytokines reduction. Angiogenesis inhibition in rats with induced asthma not only suppresses the inflammatory process through blocking VEGF expression but also inhibits the development of new blood vessels and progressing asthmatic attacks.

Topics: Angiogenesis Inhibitors; Animals; Anti-Asthmatic Agents; Asthma; Bevacizumab; Disease Models, Animal; Goblet Cells; Inflammation Mediators; Interleukin-1beta; Lung; Male; Neovascularization, Pathologic; Ovalbumin; Pneumonia; Rats, Wistar; Signal Transduction; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Asthma is a chronic inflammatory disease of the airways associated with excess production of Th2 cytokines and lung eosinophil accumulation. This inflammatory response persists in spite of steroid administration that blocks autocrine/paracrine loops of inflammatory cytokines, and the detailed mechanisms underlying asthma exacerbation remain unclear. Here, we show that asthma exacerbation is triggered by airway macrophages through a prion-like cell-to-cell transmission of extracellular particulates, including ASC protein, that assemble inflammasomes and mediate IL-1β production. OVA-induced allergic asthma and associated IL-1β production were alleviated in mice with small GTPase Arf6-deficient macrophages. The extracellular ASC specks were slightly engulfed by Arf6-/- macrophages, and the IL-1β production was reduced in Arf6-/- macrophages compared with that in WT macrophages. Furthermore, pharmacological inhibition of the Arf6 guanine nucleotide exchange factor suppressed asthma-like allergic inflammation in OVA-challenged WT mice. Collectively, the Arf6-dependent intercellular transmission of extracellular ASC specks contributes to the amplification of allergic inflammation and subsequent asthma exacerbation.

Topics: ADP-Ribosylation Factor 6; Animals; Asthma; CARD Signaling Adaptor Proteins; Cell Communication; Disease Models, Animal; Humans; Inflammasomes; Interleukin-1beta; Lung; Macrophages, Alveolar; Mice; Mice, Knockout; Ovalbumin; Phagocytosis; Symptom Flare Up; Th2 Cells; THP-1 Cells; Triazoles

Eriobotrya japonica (Thunb.) Lindl leaf (EJL) is used as a traditional Chinese medicine. E. japonica is a member of the Rosaceae family. EJL suppresses cough and relieves asthma and is widely used to treat lung diseases. In the present study, guided by the traditional Chinese medicine theory of the exterior-interior relationship between the lungs and the large intestine, the pathogenesis of cough variant asthma (CVA) and the treatment mechanism of EJL on CVA were explored.. This study aimed to explore the airway remodeling effects of EJL in CVA from the perspective of the intestinal flora and the matrix metallopeptidase 9/tissue inhibitor of metalloproteinases-1 (MMP-9/TIMP-1) pathway.. The oleanolic acid and ursolic acid contents in EJL were measured by high-performance liquid chromatography (HPLC) to ensure the quality of EJL. BALB/c mice were used to establish a CVA model through ovalbumin (OVA) sensitization and atomization. EJL (at 5, 10, or 20 g/kg/day) was intragastrically administered. The body weight, ratio of total bronchial wall area (WAt) to bronchial basement membrane perimeter (Pbm) (WAt/Pbm), the number of coughs, and cough latency were measured. The pathological changes of the lung tissue were analyzed by hematoxylin and eosin (HE) staining. The expression of α-smooth muscle actin (α-SMA) was measured by immunohistochemistry (IHC). The expressions of MMP-9 and TIMP-1 were detected in the lung tissue by reverse transcription quantitative polymerase chain reaction (RT-PCR) and Western blot analysis. Additionally, an Illumina Hiseq platform was used for 16S ribosomal DNA (16S rDNA) high-throughput sequencing to detect the intestinal flora in feces samples.. The results confirmed the positive effects of EJL on CVA. After administration of EJL, the number of coughs and the WAt/Pbm ratio decreased, the cough latency was prolonged, body weight was increased, and the general status was better than that of the CVA model mice. HE staining revealed that EJL decreased inflammatory cell infiltration and improved the histopathological structure of the lung tissue. EJL also showed significant inhibitory effects on the expression of α-SMA, MMP-9, and TIMP-1 and normalized the intestinal flora to a certain extent.. The results demonstrated that EJL alleviated airway remodeling of CVA mice, which might be related to the inhibition of the MMP-P/TIMP-1 pathway and the regulation of intestinal flora.

Topics: Animals; Asthma; Eriobotrya; Female; Gastrointestinal Microbiome; Gene Expression Regulation; Intestine, Large; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Plant Extracts; Random Allocation; Specific Pathogen-Free Organisms

Magnesium deficiency common in obesity is known to promote chronic low-grade inflammation and aggravate asthma symptoms; however, the effects of magnesium supplementation in obese asthmatic patients have not been investigated.. To examine the effects of magnesium co-administration with dexamethasone on airway inflammation in obese mice.. Female C57BL/6 mice were fed a high-fat diet, sensitized with ovalbumin (OVA) to induce allergic reactions, challenged with aerosolized OVA, and administered dexamethasone (3 mg/kg) with or without magnesium. Bronchial inflammation was analyzed based on the presence of inflammatory cells and cytokines in bronchoalveolar lavage fluid, total and OVA-specific IgE in serum, goblet cells ratios, bronchial wall thickness, and expression of α-smooth muscle actin.. In obese mice, co-administration of magnesium and dexamethasone decreased IL-13 in bronchoalveolar lavage fluid and total and OVA-specific IgE in serum, and reduced α-smooth muscle actin-positive areas in the bronchi compared with mice treated with dexamethasone alone. However, no differences were observed in dexamethasone-treated normal-weight mice depending on magnesium supplementation.. These results suggest that magnesium increases immunosuppressive effects of dexamethasone in airway inflammation aggravated by obesity, suggesting that magnesium supplementation may have a potential in alleviating asthma symptoms in obese patients with reduced responses to corticosteroids.

Topics: Adrenal Cortex Hormones; Animals; Anti-Inflammatory Agents; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Dexamethasone; Diet, High-Fat; Female; Immunoglobulin E; Immunosuppressive Agents; Inflammation; Magnesium; Mice, Inbred C57BL; Obesity; Ovalbumin

Six-week-old female BALB/c mice were sensitized and challenged with OVA in the presence and absence of chronic low-dose chlorine exposure by inhalation of naturally vaporized gas of 5% sodium hypochlorite solution. AHR, airway inflammatory cells, from BALF and the population of ILCs and macrophages in the lung were evaluated.. The mice exposed to chlorine with OVA (Cl + OVA group) showed enhanced AHR and eosinophilic inflammation compared to OVA-treated mice (OVA group). The population of T. Chronic chlorine inhalation contributes to the exacerbation of airway inflammation in asthmatic airway by mobilizing pro-inflammatory macrophage into the lung as well as stimulating group 2 and 3 ILCs.

Topics: Administration, Inhalation; Animals; Asthma; CD11 Antigens; Chlorine; Disease Models, Animal; Female; Immunity, Innate; Inflammation; Lung; Lymphocyte Count; Macrophages; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Allergic diseases and especially allergic asthma are widespread diseases with high prevalence in childhood, but also in adults. Acid sphingomyelinase (ASM) is a key regulator of the sphingolipid pathway. Previous studies defined the association of ASM with the pathogenesis of T. To determine the role of Asm under baseline conditions, wild-type (WT) and Asm. At baseline, Asm. Asm deficiency could induce higher numbers of T

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Sphingomyelin Phosphodiesterase; Th2 Cells

Acute exacerbations of asthma represent a major burden of disease and are often caused by respiratory infections. Viral infections are recognized as significant triggers of exacerbations; however, less is understood about the how microbial bioproducts such as the endotoxin (lipopolysaccharide (LPS)) trigger episodes. Indeed, increased levels of LPS have been linked to asthma onset, severity and steroid resistance.. The goal of this study was to identify mechanisms underlying bacterial-induced exacerbations by employing LPS as a surrogate for infection.. We developed a mouse model of LPS-induced exacerbation on the background of pre-existing type-2 allergic airway disease (AAD).. LPS-induced exacerbation was characterized by steroid-resistant airway hyperresponsiveness (AHR) and an exaggerated inflammatory response distinguished by increased numbers of infiltrating neutrophils/macrophages and elevated production of lung inflammatory cytokines, including TNFα, IFNγ, IL-27 and MCP-1. Expression of the type-2 associated inflammatory factors such as IL-5 and IL-13 were elevated in AAD but not altered by LPS exposure. Furthermore, AHR and airway inflammation were no longer suppressed by corticosteroid (dexamethasone) treatment after LPS exposure. Depletion of pulmonary macrophages by administration of 2-chloroadenosine into the lungs suppressed AHR and reduced IL-13, TNFα and IFNγ expression. Blocking IL-13 function, through either IL-13-deficiency or administration of specific blocking antibodies, also suppressed AHR and airway inflammation.. We present evidence that IL-13 and innate immune pathways (in particular pulmonary macrophages) contribute to LPS-induced exacerbation of pre-existing AAD and provide insight into the complex molecular processes potentially underlying microbial-induced exacerbations.

Topics: Airway Resistance; Animals; Asthma; Bacterial Infections; Bronchoalveolar Lavage Fluid; Chemokine CCL2; Cytokines; Dexamethasone; Disease Models, Animal; Disease Progression; Drug Resistance; Glucocorticoids; Interferon-gamma; Interleukin-13; Interleukins; Lipopolysaccharides; Macrophage Activation; Macrophages, Alveolar; Mice; Mucin 5AC; Ovalbumin; Respiratory Hypersensitivity; Reverse Transcriptase Polymerase Chain Reaction; Tumor Necrosis Factor-alpha

Isolated blockade of IL-25, IL-33 and thymic stromal lymphopoietin (TSLP) has been shown to reduce airways inflammation and hyperresponsiveness in murine asthma model. The hypothesis that combined blockade of all three cytokines can accomplish this more effectively has never been addressed.. We studied a murine asthma model employing sensitization and challenge with ovalbumin (OVA) or saline control. To discern the effects of IL-33 blockade, we compared outcomes in strain identical, wild-type and IL-33 receptor (St2. St2. Combined blockade of these three cytokines may better ameliorate airways pathological changes in this murine asthma model, with implications for human asthma.

Topics: Airway Remodeling; Allergens; Animals; Asthma; Cytokines; Disease Models, Animal; Eosinophils; Inflammation; Interleukin-33; Interleukins; Lung; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Thymic Stromal Lymphopoietin

Bronchial asthma is one of the most common, chronic respiratory diseases, characterized by reversible airway obstruction, eosinophil and Th2 infiltration, airway hyperresponsiveness and airway remodelling; with many cells and mediators involved. Metabolomics is a relatively new field in "omics" sciences enabling the identification of metabolome for better diagnostics and studying of diseases phenotype. The aim of this study was to investigate the role of targeted metabolomics study for better understanding of the bronchial asthma pathophysiology and finding potential biomarkers in experimental models of eosinophilic inflammation. Plasma level of 185 metabolites was measured with the AbsoluteIDQ™ p180 kit in guinea pigs with experimentally-induced allergic inflammation (n = 15) compared to naïve non-sensitised and non-challenged controls (n = 18). Of the 185 metabolites identified in plasma, 22 were significantly different and changed in ovalbumin sensitised animals. Plasma level of 13 phosphatidylcholines with saturated and unsaturated long-chain fatty acids, total phosphatidylcholines count, carnitine, symmetric dimethylarginine and its ratio to total unmodified arginine, and kynurenine to tryptophan ratio were found to be decreased, while phospholipase A2 activity indicator, tryptophan, taurine and ratio of methionine sulfoxide to unmodified methionine were found to be increased in sensitised guinea pigs compared to naïve controls. Targeted metabolomic analysis revealed significant differences in plasma metabolome of sensitised guinea pigs. Our observations point to the activation of inflammatory and immune pathways, as well as the involvement of oxidative stress.

Topics: Airway Remodeling; Allergens; Animals; Asthma; Biomarkers; Disease Models, Animal; Guinea Pigs; Lung; Male; Metabolome; Metabolomics; Ovalbumin; Oxidative Stress; Phosphatidylcholines

Myxopyrum serratulum A. W. Hill. (Oleaceae) is a traditionally used Indian medicinal plant for the treatment of cough, asthma and many other inflammatory diseases.. In this study, the protective effects of M. serratulum on airway inflammation was investigated in ovalbumin (OVA)-induced murine model of allergic asthma and lipopolysaccharide (LPS)-stimulated inflammation in RAW 264.7 murine macrophages, and the possible mechanisms were elucidated.. The phytochemicals present in the methanolic leaf extract of M. serratulum (MEMS) were identified by reverse phase high performance liquid chromatography (RP-HPLC) analysis. In vitro anti-inflammatory activity of MEMS were evaluated by estimating the levels of nitric oxide (NO), reactive oxygen species (ROS) and cytokines (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12, IL-17A, IFN-γ, TNF-α, G-CSF and GM-CSF) in LPS-stimulated RAW 264.7 macrophages. In vivo anti-asthmatic activity of MEMS was studied using OVA-induced murine model. Airway hyperresponsiveness (AHR), was measured; total and differential cell counts, eosinophil peroxidase (EPO), prostaglandin E2 (PGE2), NO, ROS, and cytokines (IL-4, IL-5 and IL-13), were estimated in bronchoalveolar lavage fluid (BALF). Serum total IgE level was measured; and the histopathological changes of lung tissues were observed. The expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in lung tissue homogenates were detected by Western blot.. The chromatographic analysis of MEMS identified the presence of gallic acid, protocatechuic acid, catechin, ellagic acid, rutin, p-coumaric acid, quercetin, naringenin and apigenin. MEMS (125 and 250 μg/mL) dose-dependently reduced the levels of NO, ROS and pro-inflammatory cytokines in LPS-stimulated RAW 264.7 macrophages. MEMS (200 and 400 mg/kg, p.o.) significantly (p < 0.05) alleviated AHR; number of inflammatory cells, EPO, PGE2, NO, ROS, and cytokines (IL-4, IL-5 and IL-13) in BALF; serum total IgE and the histopathological changes associated with lung inflammation. Western blot studies showed that MEMS substantially suppressed COX-2 and iNOS protein expressions in the lung tissues of OVA-sensitized/challenged mice.. The present study corroborates for the first time the ameliorative effects of MEMS on airway inflammation by reducing the levels of oxidative stress, pro-inflammatory cytokines and inhibiting COX-2, iNOS protein expressions, thereby validating the ethnopharmacological uses of M. serratulum.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchial Hyperreactivity; Bronchoconstriction; Cytokines; Disease Models, Animal; Female; Inflammation Mediators; Lipopolysaccharides; Lung; Macrophages; Mice; Mice, Inbred BALB C; Nitric Oxide; Oleaceae; Ovalbumin; Plant Extracts; Plant Leaves; RAW 264.7 Cells; Reactive Oxygen Species

Accumulating epidemiological studies showed that prenatal and early life exposure to ambient air pollution was important contributor to the development of childhood asthma. However, the effects and mechanisms of prenatal exposure to ozone (O. This study aimed to determine the effects and mechanism of asthma in offspring after prenatal O. Pregnant BALB/c mice were exposed to O. Airway inflammation, mucus secretion, OVA-specific immunoglobulin (Ig) E, T helper (Th) 2-skewed response, the frequency of CD3ε. Prenatal O

Topics: Air Pollutants; Animals; Asthma; Female; Killer Cells, Natural; Male; Mice, Inbred BALB C; Ovalbumin; Ozone; Pregnancy; Prenatal Exposure Delayed Effects; Th1-Th2 Balance

This study is to determine the role and mechanism of cryptotanshinone (CTS) in allergic airway inflammation. Asthma induced by OVA was established in BALB/c mice. We found increased airway hyperresponsiveness (AHR), increased inflammatory cell infiltration, elevated levels of TNF-α, interleukin-1β (IL-1β), IL-4, IL-5, IL-6 and IL-13, decreased interferon gamma (IFN-γ) in lung tissue, increased content of total immunoglobulin E (IgE), OVA specific IgE, Eotaxin, ICAM-1, VCAM-1, nuclear factor-kappaB (NF-κB) and phosphorylation of p38 MAPK in lung tissue. However, the administration of CTS significantly decreased AHR in asthmatic mice, reduced inflammation around the bronchioles and inflammatory cells around airway, regulated cytokine production, reduced the total IgE and OVA-specific IgE levels, and inhibited NF-κB activation and p38 MAPK phosphorylation. In vitro experiments in 16 HBE cells revealed that CTS attenuated CAM-1 and IL-6 expression. These results indicate that CTS alleviates allergic airway inflammation by modulating p38 MAPK phosphorylation and NF-κB activation.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Chemotaxis, Leukocyte; Cytokines; Drugs, Chinese Herbal; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Phosphorylation

Curcumin, extracted from the roots of Curcuma longa, has been used as an anti-inflammatory agent since the time of Ayurveda. The present work was designed to evaluate the potential of curcumin in amelioration of ovalbumin (OVA) induced AD in mice. Female BALB/c mice were subjected to skin OVA-patch application for a period of 1 week followed by resting period of 2 weeks, and the same protocol was repeated thrice. Curcumin was administered daily at dose of 20 mg/kg (i.p.) for 7 consecutive days during last sensitization phase. The phytochemical ameliorated the OVA-induced skin pathology as evident by normalization of epidermal thickness and suppressed infiltration of inflammatory cells in dermal region. The expression of Th2 promoting cytokines (TSLP/IL-33) and Th2 cytokines (IL-4/IL-5/IL-13/IL-31) was suppressed markedly along with reduced STAT-6 phosphorylation and GATA-3 expression. Curcumin administration also restored the redox balance and phosphorylation status of P65-NF-κB. Additionally, the epicutaneously sensitized mice challenged with aerosolized OVA developed asthmatic features which were effectively thwarted back upon curcumin treatment as reflected by data on total/differential cells in BALF and mRNA expression of Th2 cytokines in lungs. Overall, our findings demonstrate that curcumin treatment blunts the development of AD as well as associated atopic march in experimental mice.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Curcumin; Cytokines; Dermatitis, Atopic; Disease Models, Animal; Disease Progression; Female; GATA3 Transcription Factor; Lung; Mice, Inbred BALB C; Ovalbumin; Phosphorylation; Skin; STAT6 Transcription Factor; Th2 Cells; Transcription Factor RelA

In this study, we investigated whether 4-hydroxycinnamic acid (HA) has a palliative effect on asthmatic inflammatory responses using a mouse model of ovalbumin (OVA)-induced allergic asthma. The mice were divided into five groups, each consisting of seven females (normal control phosphate-buffered saline); OVA (OVA sensitization/challenge); dexamethasone (DEX, OVA sensitization/challenge + dexamethasone 3 mg/kg); HA-10 and HA-20 OVA sensitization/challenge + HA 10 and 20 mg/kg, respectively). Mice treated with HA showed a reduction in airway hyperresponsiveness and in the number of inflammatory cells in bronchoalveolar lavage fluid (BALF) compared with asthmatic control. HA treatment also reduced the levels of interleukin (IL)-5 and IL-13 in BALF and of OVA-specific immunoglobulin E in the serum compared with asthmatic control. HA treatment relieved airway inflammation and mucus overproduction caused by OVA exposure. Additionally, HA inhibited the increases in levels of nuclear factor kappa B, inducible nitric oxide synthase, and cyclooxygenase-2 that normally occur after OVA exposure. HA treatment also reduced the activity and protein level of matrix metalloproteinase-9. Taken together, HA effectively suppressed asthmatic airway inflammation and mucus production caused by OVA exposure. These findings indicate that HA has the potential to be used as a therapeutic agent for asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Coumaric Acids; Cyclooxygenase 2; Cytokines; Female; Immunoglobulin E; Inflammation; Lung; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; Mucus; Nitric Oxide Synthase Type II; Ovalbumin; Propionates; Specific Pathogen-Free Organisms

1-nitropyrene (1-NP) is widespread in the environment, as a typical nitrated polycyclic aromatic hydrocarbon. The purpose of this research was to explore the effects of gestational 1-NP exposure on susceptibility of allergic asthma in offspring. Maternal mice were exposed to 1-NP (100 μg kg

Topics: Animals; Asthma; Disease Models, Animal; Environmental Pollutants; Female; Goblet Cells; Humans; Hypersensitivity; Lung; Maternal Exposure; Mice; Mice, Inbred BALB C; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; Pyrenes

Epidemiology suggests ambient temperature is the triggers and potential activator of asthma. The role of high and low temperatures on airway inflammation of asthma, and the underlying molecular mechanism are not yet understood. A mouse model of asthma was adopted in our experiment. The BALB/c mice were exposed at different temperature for 4 h (2 h in the morning and 2 h in the afternoon) on weekday. The exposure temperatures were 10 °C, 24 °C and 40 °C. Ovalbumin (OVA) was used to sensitize the mice on days 14, 18, 22, 26, and 30, followed by an aerosol challenge for 30 min from day 32-38. After the final OVA challenge, lung function, serum protein and pulmonary inflammation were assessed. Comparing the OVA with the saline group at 24 °C, we saw a significant increase in: serum Total-IgE (p < 0.05); OVA-sIgE (p < 0.01); IL-4 (p < 0.05); IL-1β (p < 0.01); IL-6 (p < 0.01); TNF-α (p < 0.01); and the ratio of IL-4/IFN-γ (p < 0.01). At the same time, there was a significant decrease in IFN-γ (p < 0.01). As the temperature increase, there is a U shape for immune proteins and pro-inflammatory factors with a peak value at 24 °C, exception for IFN-γ (inverted U-shape). After the high and low temperature exposure, the Ri and Re increased significantly, while Cldyn decreased significantly compared with the 24 °C group. Histopathological analysis of the OVA groups showed airway remodeling, airway wall thickening and deforming, and subepithelial fibrosis. More obvious changes were found in the high and low temperature exposure groups. The immunohistochemistry suggested that TRPs changed with temperatures. High and low temperatures can aggravate airway inflammation in a mouse model of asthma. TRPs play an important role in temperature aggravation of allergic asthma. The results suggest that asthmatics should avoid exposure to high and low temperatures for too long time.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cold Temperature; Disease Models, Animal; Female; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Temperature; Tumor Necrosis Factor-alpha

Dexamethasone (DEX) is the mainstay treatment for asthma, which is a common chronic airway inflammation disease. However, the mechanism of DEX resolute symptoms of asthma is not completely clear. Here, we aimed to analyze the effect of DEX on airway inflammation in OVA-induced mice and whether this effect is related to the inhibition of the activation of NLRP3 inflammasome. Female (C57BL/6) mice were used to establish the allergic airway inflammation model by inhalation OVA. The number of inflammatory cells in the bronchi alveolar lavage fluid (BALF) was counted by Swiss-Giemsa staining, and the contents of IL-1β, IL-18, IL-5 and IL-17 were detected by ELISA. The degree of inflammatory cells infiltration and mucous cells proliferation in lung tissue were separately observed by H&E and PAS staining. The proteins expression of NLRP3, pro-caspase-1, caspase-1, IL-1β, IL-6 and IL-17 in lung tissue were detected by Western blotting. We found that DEX significantly inhibited OVA-induced inflammatory cells infiltration, airway mucus secretion and goblet cell proliferation in mice. The total and classified numbers of inflammatory cells and the levels of IL-1β, IL-18, IL-5 and IL-17 in the BALF of the experimental group were significantly lower than those of the model group after DEX treatment. DEX also significantly inhibited the activity of NLRP3 inflammasome and reduced the protein contents of Pro-Caspase-1, Caspase-1, Capase-1/Pro-Caspase-1, IL-1β, IL-6 and IL-17 in lung tissues. Our study suggested that DEX alleviates allergic airway inflammation by inhibiting the activity of NLRP3 inflammasome and the levels of IL-1β and IL-18.

Topics: Animals; Asthma; Dexamethasone; Disease Models, Animal; Female; Glucocorticoids; Humans; Inflammasomes; Interleukin-18; Interleukin-1beta; Lung; Mice; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Signal Transduction

Allergic asthma is a chronic inflammatory airway disease driven predominantly by a T. We investigated potential therapeutic effects of selective inhibition of this pathway in mice with established allergic airway disease. We further investigated whether IL-4Rα disruption in systemically sensitized mice can prevent the onset of the disease.. Inducible deletion of IL-4Rα demonstrated therapeutic effects, on established allergic airway disease, and prevented the development of ovalbumin-induced airway hyperreactivity, eosinophilia, and goblet cell metaplasia in allergen-sensitized mice. Interestingly, IL-4Rα knockdown after allergic sensitization did not induce T. Abrogation of IL-4Rα signaling after allergic sensitization would have significant therapeutic benefit for T

Topics: Allergens; Animals; Asthma; Disease Models, Animal; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

The present study explored the function and regulatory mechanism of High mobility group box 1 (HMGB1) in asthma.. OVA (ovalbumin)-induced asthmatic mice model and LPS-treated cellular model were established in this study. Airway inflammation was measured through detecting the expression of IL-4, IL-5, IL-13 and Interferon-γ (IFN-γ) in serum and BALF (bronchoalveolar lavage fluid) by ELISA kits. Bioinformatics predictive analysis, ChIP assays, Luciferase reporter assay and Western blotting were used to explore the relation between HMGB1 and HSF1 (Heat shock factor 1).. HMGB1 expression was increased in OVA-induced asthmatic mice. Silencing HMGB1 attenuated the increasing of IgE, inflammatory factors (IL-4, IL-5 and IL-13), and airway hyperresponsiveness that induced by OVA. In addition, our study found that HSF1 directly bind with the HMGB1 promoter and negatively regulation of HMGB1. HSF-1 were upregulated in OVA-induced asthmatic mice, and knockdown of HSF1 aggravated the OVA-induced airway inflammation and airway hyperreactivity in mice may through promoting the expression of HMGB1 and the activation of the Toll-like receptor 4 (TLR4)/Myeloid differentiation primary response 88 (MyD88)/Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signal pathway.. The expression of HMGB1 could be negatively regulated by HSF1, and the TLR4/MyD88/NF-κB signal pathway was involved in HSF1/HMGB1-mediated regulation of asthma.

Topics: Animals; Apoptosis; Asthma; Base Sequence; Bronchial Hyperreactivity; Cytokines; Heat Shock Transcription Factors; HEK293 Cells; HMGB1 Protein; Humans; Inflammation; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Myeloid Differentiation Factor 88; NF-kappa B; Ovalbumin; Promoter Regions, Genetic; Signal Transduction; Toll-Like Receptor 4

Allergies affect a significant proportion of the world's population, and existing vaccination strategies to restrict their adverse pathologies often render side-effects. The aim of this study was to design a new vaccine for allergen-specific immunotherapy (SIT), and to investigate its preventive effects during allergic inflammation. We constructed ovalbumin (OVA)-conjugated celastrol-loaded nanomicelles (OVA-NMs-celastrol), wherein celastrol (a bioactive anti-inflammatory compound) was loaded into carboxyl-functioned polymeric nanomicelles using a thin-film hydration method. OVA was used as a model allergen and conjugated on nanomicelles. The OVA-NMs-celastrol obtained were characterized based on particle size, morphology, drug encapsulation efficiency, and drug loading percentage. Further, the preventive effect of OVA-NMs-celastrol was evaluated in a mouse model of allergic asthma. Our results showed that OVA-NMs-celastrol possessed valuable characteristics such as small particle size (50.72 ± 0.98 nm) and spherical-like shape, with celastrol encapsulation efficiency of 99.89 ± 0.85% and a drug loading percentage of 4.76 ± 0.03%. Further, in vivo results showed that treatment with OVA-NMs-celastrol could decrease OVA specific IgE and histamine levels, Th2 cytokine (IL-4, IL-5) levels, and inflammatory cell infiltration in the lung tissues. Moreover, it could enhance the OVA specific IgG1 and IgG2a levels and decrease the IgE / IgG2a ratio. These results demonstrate the successful construction of OVA-NMs-celastrol as a potential vaccine candidate for use in SIT for allergic inflammation.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Eosinophils; Female; Histamine Release; Immunoglobulin E; Immunoglobulin G; Lung; Mice, Inbred BALB C; Micelles; Nanostructures; Ovalbumin; Pentacyclic Triterpenes; Triterpenes

Allergic airway inflammation is one of the major pathological events involved in asthma, and dysregulation of regulatory T cells (Treg) plays a crucial role in the development of allergic airway inflammation. Here, we attempted to investigate the regulatory effects of B cell-activating factor (BAFF) on Tregs in allergic airway inflammation.. BAFF expression was analyzed by ELISA, quantitative reverse transcription PCR (RT-PCR) and Western blot assays. The levels of IL-4, TGF-β, IL-2, and IL-10 were tested using ELISA kits. Flow cytometry was conducted to analyze the populations of CTLA4. BAFF was found to be aberrantly expressed in sputum and lungs in patients with asthma as well as OVA sensitized mice. BAFF silencing by lentiviral BAFF shRNA reduced the number of eosinophils and levels of IL-4 in the BAL fluid, as well as the Fizz1 expression in the lungs of OVA mice. Additionally, the population of CTLA4. BAFF has an inhibitory effect on the generation and suppressor function of Tregs by affecting pro-Tregs cytokines, thereby contributing to the development of allergic airway inflammation.

Topics: Adult; Animals; Asthma; B-Cell Activating Factor; Case-Control Studies; Cytokines; Female; Gene Expression Regulation; Gene Silencing; Humans; Inflammation; Male; Mice; Mice, Inbred BALB C; Middle Aged; Ovalbumin; T-Lymphocytes, Regulatory

Skewed T helper (Th)2 response plays a crucial role in the pathogenesis of allergic diseases. The therapeutic efficacy for allergic diseases is unsatisfactory currently. This study aims to regulate the skewed Th2 response with CARsomes.. The CARsome consisted of an epitope of Dermatophagoides farina-1 (Derf1), a segment of the anti-DEC205 antibody, the scFv, and an open reading frame of perforin. This fusion protein binds to DEC205 molecule on the surface of exosomes derived from dendritic cells (DC). The effects of CARsome on inducing antigen (Ag)-specific Th2 cell apoptosis were assessed both in vivo and in vitro.. Exposure to CARsomes in the culture induced Ag-specific Th2 cell apoptosis. Injection of CARsomes through the vein puncture also induced Ag-specific Th2 cell apoptosis in the lungs of sensitized mice. CARsomes could induce Ag-specific regulatory T cells. Administration of CARsomes efficiently inhibited experimental allergic airway inflammation.. The CARsomes can inhibit allergic airway inflammation by inducing Ag-specific Th2 cell apoptosis and induce Ag-specific regulatory T cells. The data suggest that CARsomes have the translational potential to be used to treat allergic airway inflammation.

Topics: Animals; Antigens; Apoptosis; Asthma; Dendritic Cells; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Airway remodeling in asthma is difficult to treat because of its complex pathophysiology that involves proinflammatory cytokines, as well as the arachidonic acid cytochrome P-450 (CYP) pathway; however, it has received little attention. In this study, we assessed the efficacy of a soluble epoxide hydrolase (sEH) on airway remodeling in a mouse model of chronic asthma. The expression of sEH and CYP2J2 and the level of 14,15-epoxyeicosatrienoic acid (14,15-EET), airway remodeling and hyperresponsiveness (AHR) were analyzed to determine the level of sEH inhibition. AUDA, a sEH inhibitor, was given daily for 9 weeks orally, which significantly increased the level of 14,15-EET by inhibiting the expression of sEH and increasing the expression of CYP2J2 in lung tissues. The inhibition of sEH reduced the expression of remodeling-related molecular markers, such as interleukin (IL)-13, IL-17, matrix metalloproteinase 9, N-cadherin, α-smooth muscle actin (α-SMA), S100A4, Twist, epithelial goblet cell metaplasia, and collagen deposition in bronchoalveolar lavage fluid (BAL fluid) and lung tissues. Moreover, remodeling-related eosinophil accumulation in the BAL fluid and infiltration into the lung tissue were improved by AUDA. Finally, AUDA alleviated AHR, which is a functional indicator of airway remodeling. The effect of AUDA on airway remodeling was related to the downregulation of extracellular-regulated protein kinases (Erk1/2), c-Jun N-terminal kinases (JNK) and signal transducer and activator of transcription 3 (STAT3). To our knowledge, this is the first report to demonstrate that inhibition of sEH exerts significant protective effects on airway remodeling in asthma.

Topics: 8,11,14-Eicosatrienoic Acid; Adamantane; Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytochrome P-450 CYP2J2; Cytochrome P-450 Enzyme System; Disease Models, Animal; Epoxide Hydrolases; Female; Humans; Lauric Acids; Lung; MAP Kinase Signaling System; Mice; Ovalbumin; Signal Transduction; STAT3 Transcription Factor

Exposure to PM. Fifty rats (25 males and 25 females) were divided randomly into the control group, ovalbumin asthmatic model group, asthma low-, middle- and high-dose groups (n = 10, 5 males and 5 females each group). The control group, ovalbumin asthmatic model group received physiological saline; the asthma low-, middle- and high-dose groups received 1.5, 7.5 and 37.5 mg/kg PM. Respiratory mucus secretion increased with increasing dose of PM. Short-term exposure to a high concentration of PM

Topics: Air Pollutants; Animals; Asthma; Female; Inflammation; Lung; Male; Mucus; NF-kappa B; NF-KappaB Inhibitor alpha; Ovalbumin; Particulate Matter; Rats; Signal Transduction; Tumor Necrosis Factor-alpha

Propolis is a natural product produced by honeybees used as a medicine at least to 300 BC. In the last decades, several studies showed biological and pharmacological properties of propolis, witch scientifically explains the empirical use for centuries. The anti-inflammatory activity of propolis with the purpose to reduce Th2 inflammation has been evaluated in allergic asthma. However, it remains to be determined how propolis negatively regulates the immune response after allergen re-exposure.. We hypothesized that the anti-inflammatory activity of propolis is dependent on the induction of myeloid derived suppressor cells (MDSC) and regulatory T cells.. To assess this hypothesis, we used an ovalbumin-induced asthma model to evaluate the effect of EPP-AF® dry extract from Brazilian green propolis.. Propolis treatment decreased pulmonary inflammation and mucus production as well as eosinophils and IL-5 in the broncoalveolar lavage. Propolis enhanced also in vitro differentiation and in vivo frequency of lung MDSC and CD4. Together these results confirm the immunomodulatory potential of propolis during sensitization and challenge with allergen. In addition, the collecting findings show, for the first time, that propolis increases the frequency of MDSC and CD4

Topics: Allergens; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cell Differentiation; Female; Immunologic Factors; Immunotherapy; Interleukin-13; Interleukin-5; Lung; Mice, Inbred C57BL; Myeloid-Derived Suppressor Cells; Ovalbumin; Propolis; T-Lymphocytes, Regulatory; Th2 Cells

Asthma is a complex inflammatory disease which affects multiple individuals worldwide especially pediatric ages.. This study aimed to assess the possible protective effect of carvacrol, as natural antioxidant anti-inflammatory drug, against bronchial asthma induced experimentally in rats.. Rats were randomly allocated into 5 groups; a normal control group, control drug group received only carvacrol, an asthma control group, a standard treatment group receiving dexamethasone (DEXA) and carvacrol treatment group. Bronchial asthma was induced by sensitization with i.p dose followed by challenge with intranasal dose of ovalbumin (OVA). 24 h after the last challenge, absolute eosinophil count (AEC) were determined in bronchoalveolar lavage fluids (BALF). Immunoglobulin E (IgE) was determined in serum. Inflammatory biomarkers like Interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin 13 (IL-13), tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) were also measured in BALF. Nitrosative stress biomarker namely inducible nitric oxide synthase (iNOS) was determined in BALF as well as oxidative stress biomarkers namely superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) were determined in lung tissue. Additionally, histopathological study, immunohistochemical study of UCN and western blot analysis of SP-D were performed.. Carvacrol administration significantly reduced the values of AEC, IgE, IL-4, IL-5, IL-13, TNF-α, IFN-γ, iNOS and MDA, while it significantly increased the values of SOD and GSH as compared to the asthmatic group. Histopathological, immunohistochemical and western blot study reinforced the biochemical results.. Carvacrol may be a promising protective agent against bronchial asthma induced experimentally in rats.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Blotting, Western; Cymenes; Disease Models, Animal; Immunologic Factors; Interleukin-13; Interleukin-4; Interleukin-5; Male; Nitric Oxide Synthase Type II; Ovalbumin; Pulmonary Surfactant-Associated Protein D; Rats; Rats, Wistar; Real-Time Polymerase Chain Reaction

In this study, we investigated the role and mechanism of imperatorin (IMP) in chronic inflammation and airway remodeling. The levels of TNF-α, IL-1β, IL-6, IL-8, VEGF, α-SMA, and ROS were detected by ELISA, immunohistochemistry (IHC), immunofluorescence, and Western blot. In addition, we evaluated the effect of IMP on MAPK, PI3K/Akt, NF-κB, and Nrf2/HO-1 signaling pathways. IMP treatment obviously attenuated the production of inflammatory cytokines and inflammatory cells in bronchoalveolar lavage fluid of OVA-induced airway remodeling model. Meanwhile, it significantly inhibited inflammatory cell infiltration, goblet cell hyperplasia, collagen deposition, VEGF production, α-SMA, and ROS expression. Our study has shown that IMP could regulate the signaling pathways including MAPK, PI3K/Akt, NF-κB, and Nrf2/HO-1 to release the inflammatory responses. IMP might attenuate airway remodeling by the down-regulation of Nrf2/HO-1/ROS/PI3K/Akt, Nrf2/HO-1/ROS/MAPK, and Nrf2/HO-1/ROS/NF-κB signaling pathways.

Topics: Airway Remodeling; Animals; Asthma; Cell Line; Cytokines; Disease Models, Animal; Female; Furocoumarins; Heme Oxygenase-1; Membrane Proteins; Mice; Mice, Inbred BALB C; Myocytes, Smooth Muscle; NF-E2-Related Factor 2; Ovalbumin; Reactive Oxygen Species; Signal Transduction

Scrophularia buergeriana Miq. (Scrophulariaceae) (SB) has been used as an oriental medicine for the treatment of inflammatory diseases, such as neuritis and pharyngolaryngitis.. We explored the therapeutic effects of S. buergeriana ethanol extract (SBE) on airway inflammation in ovalbumin (OVA)-induced asthmatic mice and lipopolysaccharide (LPS)-stimulated RAW264.7 cells.. Mice were intraperitoneally injected with OVA on days 0 and 14 to elevate the immune response. On days 21 to 23, the mice were challenged with OVA solution and SBE (20 and 40 mg/kg) was administered daily by oral gavage from days 18 to 23. RAW264.7 cells were pretreated with SBE 1 h before LPS stimulation.. SBE administration effectively suppressed inflammatory cell infiltration, the expression of interleukin (IL)-5, IL-13, and IL-17, immunoglobulin E, and airway hyperresponsiveness in an OVA-induced allergic asthma model. A reduction in histological alterations, including airway inflammation and mucus hypersecretion, was observed. These effects of SBE were accompanied by a decrease in matrix metalloproteinase-9 (MMP-9) expression and nuclear factor kappa B (NF-κB) phosphorylation. These responses were observed in LPS-stimulated RAW264.7 cells. SBE treatment reduced the mRNA expression of tumor necrosis factor (TNF)-α, IL-6, and MMP-9, and NF-κB phosphorylation, in LPS-stimulated RAW264.7 cells.. Our results indicated that SBE effectively attenuated airway inflammation in an OVA-induced allergic asthma model. These properties of SBE were thought to be involved in the suppression of NF-κB phosphorylation, suggesting that the material has the potential to regulate the development of allergic asthma.

Topics: Animals; Asthma; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lipopolysaccharides; Matrix Metalloproteinase 9; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Phosphorylation; Plant Extracts; RAW 264.7 Cells; Scrophularia

To assess the efficacy of the novel clinical formulation of fenretinide (LAU-7b) for the treatment of allergic asthma. To study the association between LAU-7b treatment in allergic asthma and the modulation of very long chain ceramides (VLCC).. We used two allergens (OVA and HDM) to induce asthma in mouse models and we established a treatment protocol with LAU-7b. The severity of allergic asthma reaction was quantified by measuring the airway resistance, quantifying lung inflammatory cell infiltration (Haematoxylin and eosin stain) and mucus production (Periodic acid Schiff satin). IgE levels were measured by ELISA. Immunophenotyping of T cells was done using Fluorescence-activated cell sorting (FACS) analysis. The analysis of the specific species of lipids and markers of oxidation was performed using mass spectrometry.. Our data demonstrate that 10 mg/kg of LAU-7b was able to protect OVA- and HDM-challenged mice against increase in airway hyperresponsiveness, influx of inflammatory cells into the airways, and mucus production without affecting IgE levels. Treatment with LAU-7b significantly increased percentage of regulatory T cells and CD4. 9 days of 10 mg/kg of LAU-7b daily treatment protects the mice against allergen-induced asthma and restores VLCC levels in the lungs and plasma.

Topics: Allergens; Animals; Asthma; Ceramides; Clinical Protocols; Disease Models, Animal; Drug Compounding; Female; Fenretinide; Male; Methylcellulose; Mice; Ovalbumin; Pyroglyphidae

The skewed T helper (Th) 2 response plays a critical role in the pathogenesis of allergic asthma. Regulatory T (Treg) cells and the regulatory cytokines are required in maintaining the homeostasis in the body. This study aims to determine the effects of a poly(lactic-co-glycolic) acid (PLGA)-ovalbumin (OVA)+A20 (a ubiquitin E3 ligase) nanovaccine on inhibiting allergic asthma in a murine model. In this study, A20 and OVA (a model antigen) were encapsulated into PLGA to be a nanovaccine (PLGA-OVA+A20). An allergic asthma murine model was developed with OVA as the specific antigen to test the role of PLGA-OVA+A20 nanovaccine in maintaining the immune homeostasis in the airway tissues. The results showed that PLGA-OVA+A20 nanovaccine inhibited the asthma responses in mice by suppressing Th2 inflammatory responses, promoting the generation of Treg cells in the airway tissues. We conclude that the PLGA-OVA+A20 nanovaccine has a marked inhibitory effect on the airway allergic response in sensitized mice by significantly promoting the generation of Treg cell and IL-10. The data suggest that PLGA-OVA+A20 has translational potential in the treatment of allergic asthma.

Topics: Animals; Asthma; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Mice; Mice, Inbred BALB C; Nanoparticles; Ovalbumin; Polylactic Acid-Polyglycolic Acid Copolymer; T-Lymphocytes, Regulatory; Tumor Necrosis Factor alpha-Induced Protein 3

In allergen-specific immunotherapy for asthma, antigens attached to dendritic cells increase the tryptophan metabolism in these cells and alter the Th17/Treg balance in the airways. Tryptophan metabolism has long been suggested to be relevant in the pathophysiology of allergic disorders, including asthma. Our study investigated whether tryptophan metabolites are responsible for the changes in Th17/Treg balance and decreases in airway hyperreactivity and inflammation seen during allergen-specific immunotherapy in an asthma model. Ovalbumin was injected intraperitoneally into mice to establish an asthma model, and then high dose ovalbumin allergen-specific immunotherapy was administered to induce immune tolerance. Airway hyperreactivity and serum ovalbumin-specific immunoglobulin E were measured to assess whether the animal model was successfully established. We then examined the influence of inhibition of tryptophan metabolism and the addition of tryptophan metabolites on allergen-specific immunotherapy-induced changes in the Th17/Treg balance and decreases in airway inflammation and inflammatory cytokines. Production of tryptophan metabolites was partly responsible for the allergen-specific immunotherapy-induced increase in Tregs, decrease in airway inflammation, and decrease in inflammatory cytokines. Ovalbumin-specific immunoglobulin E and airway hyperreactivity were not affected. In the context of asthma, an increase in tryptophan metabolites is one of the mechanisms by which allergen-specific immunotherapy achieves immune tolerance.

Topics: Allergens; Animals; Asthma; Cytokines; Disease Models, Animal; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Tryptophan

IL-33 and its receptor ST2 are contributing factors to airway inflammation and asthma exacerbation. The IL-33/ST2 signaling pathway is involved in both the onset and the acute exacerbations of asthma. In this study, we address the role of endogenous IL-33 and its autoamplification of the IL-33/ST2 pathway in Ag-dependent and Ag-independent asthma-like models. Wild-type, IL-33 knockout, ST2 knockout mice were either intratracheally administrated with 500 ng of rIL-33 per day for four consecutive days or were sensitized and challenged with OVA over 21 d. In wild-type mice, IL-33 or OVA induced similar airway hyperresponsiveness and eosinophilic airway inflammation. IL-33 induced its own mRNA and ST2L mRNA expression in the lung. IL-33 autoamplified itself and ST2 protein expression in airway epithelial cells. OVA also induced IL-33 and ST2 protein expression. In IL-33 knockout mice, the IL-33- and OVA-induced airway hyperresponsiveness and eosinophilic airway inflammation were both significantly attenuated, whereas IL-33-induced ST2L mRNA expression was preserved, although no autoamplification of IL-33/ST2 pathway was observed. In ST2 knockout mice, IL-33 and OVA induced airway hyperresponsiveness and eosinophilic airway inflammation were both completely diminished, and no IL-33/ST2 autoamplification was observed. These results suggest that endogenous IL-33 and its autoamplification of IL-33/ST2 pathway play an important role in the induction of asthma-like phenotype. Thus an intact IL-33/ST2 pathway is necessary for both Ag-dependent and Ag-independent asthma-like mouse models.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Epithelial Cells; Humans; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Mice; Mice, Knockout; Ovalbumin; Recombinant Proteins; Respiratory Mucosa; Signal Transduction

Asthma is a chronic inflammatory disease that represents high hospitalizations and deaths in world. Copaiba oil (CO) is popularly used for relieving asthma symptoms and has already been shown to be effective in many inflammation models. This study aimed to investigate the immunomodulatory relationship of CO in ovalbumin (OVA)-induced allergic asthma. The composition of CO sample analyzed by GC and GC-MS and the toxicity test was performed in mice at doses of 50 or 100 mg/kg (by gavage). After, the experimental model of allergic asthma was induced with OVA and mice were orally treated with CO in two pre-established doses. The inflammatory infiltrate was evaluated in bronchoalveolar lavage fluid (BALF), while cytokines (IL-4, IL-5, IL-17, IFN-γ, TNF-α), IgE antibody and nitric oxide (NO) production was evaluated in BALF and lung homogenate (LH) of mice, together with the histology and histomorphometry of the lung tissue. CO significantly attenuated the number of inflammatory cells in BALF, suppressing NO production and reducing the response mediated by TH2 and TH17 (T helper) cells in both BALF and LH. Histopathological and histomorphometric analysis confirmed that CO significantly reduced the numbers of inflammatory infiltrate in the lung tissue, including in the parenchyma area. Our results indicate that CO has an effective in vivo antiasthmatic effect.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Dose-Response Relationship, Drug; Fabaceae; Female; Humans; Lung; Mice; Nitric Oxide; Oils, Volatile; Ovalbumin; Plant Oils; Th17 Cells; Th2 Cells; Toxicity Tests, Acute

Asthma outcomes is aggravated in obese patients. Excess of methylglyoxal (MGO) in obese/diabetic patients has been associated with diverse detrimental effects on cell function. This study aimed to evaluate the effects of long-term oral intake of MGO on ovalbumin-induced eosinophil inflammation. Male C57/Bl6 mice received 0.5% MGO in the drinking water for 12 weeks. Mice were sensitized and challenged with ovalbumin (OVA), and at 48 h thereafter, bronchoalveolar lavage (BAL) fluid and lungs were collected for cell counting, morphological analysis, and ELISA, mRNA expressions and DHE assays. In MGO-treated mice, OVA challenge significantly increased the peribronchiolar infiltrations of inflammatory cells and eosinophils compared with control group. Higher levels of IL-4, IL-5, and eotaxin in BAL fluid were also detected in MGO compared with control group. In addition, lung tissue of MGO-treated mice displayed significant increases in mRNA expressions of NF-κB and iNOS whereas COX-2 expression remained unchanged. The high TNF-α mRNA expression observed in lungs of OVA-challenged control mice was not further increased by MGO treatment. In MGO group, OVA-challenge increased significantly the NOX-2 and NOX-4 mRNA expressions, without affecting the NOX-1 expression. Levels of reactive-oxygen species (ROS) were significantly higher in lungs of MGO-treated mice, and no further increase by OVA-challenge was observed. In conclusion, 12-week intake of MGO exacerbates Th2-mediated airway eosinophil infiltration by activation of NF-kB/iNOS-dependent signaling pathway and positive regulation of NOX-2 and NOX-4 in the lung tissues. Scavengers of MGO could be an option to prevent obesity-related asthma.

Topics: Allergens; Animals; Asthma; Cell Movement; Disease Models, Animal; Eosinophils; Humans; Interleukin-4; Interleukin-5; Male; Mice; Mice, Inbred C57BL; NADPH Oxidase 4; NF-kappa B; Obesity; Ovalbumin; Pyruvaldehyde; Reactive Oxygen Species; Signal Transduction; Th2 Cells

Patients with late-onset asthma (LOA) have poor clinical outcomes. Osteopontin (OPN) is associated with airway inflammation and remodeling. To investigate the role of OPN in LOA compared to early-onset asthma (EOA), serum OPN levels were compared between 131 adult asthma patients (48 LOA and 83 EOA patients) and 226 healthy controls (HCs). BALB/c mice were sensitized with ovalbumin with/without polyinosinic-polycytidylic acid (poly(I:C)) from week 6 (A6 mice) or week 12 (A12 mice) after birth. Airway hyperresponsiveness (AHR), bronchoalveolar lavage fluid (BALF), cell counts, histology, and Spp1 expression were assessed. The levels of OPN, transforming growth factor β1 (TGF-β1), chitinase 3-like 1 (CH3L1), and interleukin (IL) 5 were measured by ELISA. The expression of Smad3 phosphorylation and tissue transglutaminase 2 (TGM2) was evaluated by Western blot. The serum OPN levels were significantly higher in asthma patients than in HCs and in LOA patients than in those with EOA (P < 0.05) and were positively correlated with serum TGF-β1 and CH3L1 (r = 0.174, r = 0.264; P < 0.05). A12 mice showed elevated AHR with increased levels of OPN/TGF-β1/IL-5 in BALF and Spp1 compared to A6 mice. Poly(I:C) induced remarkable TGF-β1, CH3L1, Th2 cytokine, and OPN levels in BALF and the expression of phosphorylated Smad3, TGM2, and Spp1 in the lungs. OPN triggered TGF-β1/Smad3 signaling in the lungs, which was suppressed by dexamethasone and anti-IL5 antibody. In conclusion, aging and exposure to viral infections may induce OPN release and consequently modulate inflammation and TGF-β1/Smad3-related remodeling, contributing to the development of LOA.

Topics: A549 Cells; Adult; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Line, Tumor; Cytokines; Female; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Osteopontin; Ovalbumin; Phenotype; Protein Glutamine gamma Glutamyltransferase 2; Transforming Growth Factor beta1

This study aimed to investigate the functions of the amygdala in rat asthma model.. Wheat germ agglutinin-horseradish peroxidase (WGA-HRP) was used for tracing from the paraventricular nucleus (PVN) to the amygdala, and nuclear lesions were performed to observe changes in respiratory function and airway inflammation.. This study showed that the extracellular neuronal discharged in the medial amygdala (MeA) and central amygdala (CeA), and the expression of Fos significantly increased in asthmatic rat compared to control group. The distribution of Fos- and oxytocin (OT)-positive neurons and Fos/OT dual-positive neurons evidently increased in the PVN. WGA-HRP was injected into the PVN for tracing, and Fos/HRP-dual-positive neurons were observed to be distributed in the MeA. By using kainic acid (KA) to injure the MeA and CeA in asthmatic rats, expiratory and inspiratory times (TE/TI) and airway resistance (Raw) decreased, and minute ventilation volume (MVV) and dynamic pulmonary compliance (Cdyn) increased accordingly. In the bronchoalveolar lavage fluid (BALF), the number of eosinophils and the concentration of IL-4 were lower than those of the control group, and the ratio of Th1/Th2 cells was higher than that of the control group. In the PVN, the distribution of Fos-, OT-positive cells and Fos/OT double-positive cells decreased compared with those of the control group. The activities of the MeA and CeA and of OT neurons in the PVN of the rats were correlated with the occurrence of asthma.. Asthma attack could induce neural activities in the MeA and CeA, and OT neurons in the PVN may be involved in the process of asthma attack.

Topics: Amygdala; Animals; Asthma; Male; Ovalbumin; Paraventricular Hypothalamic Nucleus; Rats; Rats, Sprague-Dawley

Allergic airway diseases are accompanied by increased permeability and an inflammatory state of epithelial barriers, which are thought to be susceptible to allergen sensitization. Although exogenous drivers (proteases, allergens) of epithelial barrier disruption and sensitization are well studied, endogenous contributors (diet, xenobiotics, hormones, and metabolism) to allergic sensitization are much less understood. Xenoestrogens are synthetic or natural chemical compounds that have the ability to mimic estrogen and are ubiquitous in the food and water supply of developed countries. By interfering with the estrogen produced by the endocrine system, these compounds have the systemic potential to disrupt the homeostasis of multiple tissues. Our study examined the potential of prototypical xenoestrogen bisphenol A (BPA) to disrupt epithelial homeostasis

Topics: Administration, Inhalation; Animals; Asthma; Benzhydryl Compounds; Cell Line; Cell Proliferation; Cytokines; Drug Hypersensitivity; Endocrine Disruptors; Epithelial Cells; Female; Gene Expression Regulation; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phenols; Respiratory Mucosa

Epidemiologic studies show that there is a link between Bisphenol A (BPA) exposure and lung inflammation. Despite this, the molecular mechanisms are not entirely known. This study sought to determine whether exposure to BPA affected the development of ovalbumin (OVA) induced lung inflammation in adolescent female mice and whether the mechanism was related to mTOR-mediated autophagy pathway. Female 4-week-old C57BL/6 mice after one week of domestication were randomly divided into five groups (8/group): control group, OVA group, 0.1 μg mL

Topics: Animals; Asthma; Autophagy; Benzhydryl Compounds; Cytokines; Environmental Pollutants; Female; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; Phenols; Pneumonia; Respiratory Hypersensitivity; TOR Serine-Threonine Kinases; Toxicity Tests

Mechanisms mediating the protective effects of molecular hydrogen (H

Topics: Animals; Asthma; Bronchoconstrictor Agents; Cells, Cultured; Disease Models, Animal; Female; Glycolysis; Humans; Hydrogen; Lactic Acid; Leukocytes, Mononuclear; Lung; Male; Methacholine Chloride; Mice; Middle Aged; Ovalbumin; Oxidative Phosphorylation; Primary Cell Culture

DNA damage could lead to the accumulation of cytosolic DNA, and the cytosolic DNA-sensing pathway has been implicated in multiple inflammatory diseases. However, the role of cytosolic DNA-sensing pathway in asthma pathogenesis is still unclear. This article explored the role of airway epithelial cyclic GMP-AMP synthase (cGAS), the major sensor of cytosolic dsDNA, in asthma pathogenesis. Cytosolic dsDNA accumulation in airway epithelial cells (ECs) was detected in the setting of allergic inflammation both in vitro and in vivo. Mice with cGAS deletion in airway ECs were used for OVA- or house dust mite (HDM)-induced allergic airway inflammation. Additionally, the effects of cGAS knockdown on IL-33-induced GM-CSF production and the mechanisms by which IL-33 induced cytosolic dsDNA accumulation in human bronchial epithelial (HBE) cells were explored. Increased accumulation of cytosolic dsDNA was observed in airway epithelium of OVA- or HDM-challenged mice and in HBE cells treated with IL-33. Deletion of cGAS in the airway ECs of mice significantly attenuated the allergic airway inflammation induced by OVA or HDM. Mechanistically, cGAS participates in promoting T

Topics: Airway Remodeling; Allergens; Animals; Antigens, Dermatophagoides; Asthma; Cytosol; Dermatophagoides pteronyssinus; DNA Damage; DNA, Mitochondrial; Epithelial Cells; Gene Knockdown Techniques; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Interleukin-33; Mice; Mice, Transgenic; Mitochondria; Nucleotidyltransferases; Ovalbumin; Respiratory Mucosa

Topics: Allergens; Animals; Anti-Bacterial Agents; Asthma; Azithromycin; Bronchoalveolar Lavage Fluid; Cefixime; Cytokines; Fecal Microbiota Transplantation; Feces; Female; Gastrointestinal Microbiome; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Airway remodeling is a key feature of asthma. Extracellular matrix synthesis and vascular remodeling respectively regulated by transforming growth factor (TGF-β1) and vascular endothelial growth factor (VEGF), are important for the airway remodeling. This study aimed to investigate the effect of Soufeng Yuchuan (SFYC) decoction, a Traditional Chinese Medicine, on airway remodeling and expression of VEGF and TGF-β1 in asthma model rats. A rat model of asthma was induced by ovalbumin (OVA) treatment. The results showed that SFYC decoction improved general conditions and reduced the damage in lung tissues in asthma model rats. Furthermore, SFYC decoction significantly reduced the OVA-induced levels of VEGF and TGF-β1 in sera and in bronchoalveolar lavage fluid. Moreover, SFYC decoction decreased the OVA-induced VEGF mRNA and protein levels in lung tissues in asthma model rats. Interestingly, SFYC with high dose was more potent in reducing TGF-β1 level in rat sera and BALF than dexamethasone (positive control). In summary, SFYC decoction effectively mitigates lung damage in OVA-induced asthma model rats, which was associated with inhibition of VEGF and TGF-β1.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Dexamethasone; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Male; Ovalbumin; Rats; Rats, Sprague-Dawley; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

Allergic asthma is a chronic inflammatory disease characterized by airflow obstruction, airway hyperresponsiveness (AHR), airway inflammation, and mucus overproduction. Cordyceps polysaccharide (CPS) is one of the main bioactive compounds of Cordyceps militarisis, a traditional Chinese medicine. In this study, we established a mouse model of asthma using ovalbumin (OVA) challenge and evaluated the potential regulatory effect of CPS (25, 50, and 100 mg/kg) on asthmatic mice. These results showed that the asthmatic mice treated with CPS suppressed the secretion of eotaxin, IL-4, IL-5, IL-13, and IFN-γ in the blood and bronchoalveolar lavage fluid (BALF), and decreased serum IgE levels compared to the vehicle-treated mice. CPS also alleviated inflammatory cell infiltration, goblet cell hyperplasia, and the increases of inflammatory cells in the mouse model of asthma. In addition, OVA-induced AHR was inhibited by CPS treatment. Further analyses of protein expression revealed that CPS inhibited the activation of transforming growth factor β1 (TGF-β1)/Smad pathway in mice with asthma. These findings indicated that CPS might serve as a potential therapeutic agent for the management of allergic asthma.

Topics: Animals; Asthma; Cordyceps; Fungal Polysaccharides; Interferon-gamma; Interleukin-13; Interleukin-4; Interleukin-5; Lung; Medicine, Chinese Traditional; Mice; Ovalbumin; Respiratory Hypersensitivity; Signal Transduction; Smad2 Protein; Smad3 Protein; Transforming Growth Factor beta1

Acetyl CoA carboxylase (ACC) regulates the differentiation of Th1, Th2, Th17 cells and Treg cells, which play a critical role in airway inflammation of asthma. Here we investigated the role of ACC in the pathogenesis of asthma.. Chicken Ovalbumin-sensitized and -challenged mice were divided into three groups, PBS group, DMSO (solvent of TOFA) group and ACC inhibitor 5-tetradecyloxy-2-furoic acid (TOFA) + DMSO group. Airway inflammation was assessed with histology, percentages of CD4. In asthma mice, the expression of ACC increased, while the expression of phosphorylated ACC (pACC) decreased. TOFA had no significant effect on pACC expression. TOFA reduced serum IgE, airway inflammatory cells infiltration and goblet cell hyperplasia, but dramatically increased airway responsiveness. TOFA significantly reduced the percentages of Th1, Th2, Th17 cells in lung and spleen, the expression of GATA3 and RORγt in lung, and IFN-γ, IL-4, IL-17A levels in BALF and serum. TOFA had no significant effect on the percentage of Treg cells, IL-10 level and the expression of T-bet and Foxp3.. Acetyl-CoA carboxylase inhibitor TOFA might have a distinct effect on asthmatic airway inflammation and airway hyperresponsiveness.

Topics: Acetyl-CoA Carboxylase; Airway Resistance; Animals; Asthma; Chickens; Disease Models, Animal; Female; Furans; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; Respiratory Hypersensitivity; Treatment Outcome

Topics: 1-Methyl-3-isobutylxanthine; Administration, Inhalation; Airway Resistance; Animals; Asthma; Bronchoalveolar Lavage Fluid; Chemokine CCL2; Disease Models, Animal; Humans; Lipopolysaccharides; Male; Mice; Nebulizers and Vaporizers; Ovalbumin; Phosphodiesterase Inhibitors; Phosphoric Diester Hydrolases; Pneumonia; Signal Transduction; Specific Pathogen-Free Organisms; Tidal Volume

Innate immune activation through exposure to indoor and outdoor pollutants is emerging as an important determinant of asthma severity. For example, household levels of the bacterial product lipopolysaccharide (LPS) are associated with increased asthma severity. We hypothesized that activation of the innate immune receptor TLR5 by its bacterial ligand flagellin will exacerbate airway inflammation and asthma symptoms.. We determined the effect of flagellin co-exposure with ovalbumin in a murine model of allergic asthma. We evaluated the presence of flagellin activity in house dust of asthma patients. Finally, we analyzed the association of a dominant-negative polymorphism in TLR5 (rs5744168) with asthma symptoms in patients with asthma.. We showed that bacterial flagellin can be found in the house dust of patients with asthma and that this bacterial product exacerbates allergic airway inflammation in an allergen-specific mouse model of asthma. Furthermore, a dominant-negative genetic polymorphism in TLR5, the receptor for flagellin, is associated with decreased symptoms in patients with asthma.. Together, our results reveal a novel genetic protective factor (TLR5 deficiency) and a novel environmental pollutant (microbial flagellin) that influence asthma severity. (Clinical trials NCT01688986 and NCT01087307).

Topics: Adult; Animals; Asthma; Bronchial Hyperreactivity; Bronchoconstriction; Case-Control Studies; Cross-Sectional Studies; Cytokines; Disease Models, Animal; Female; Flagellin; HEK293 Cells; Humans; Lung; Male; Mice, Inbred C57BL; Middle Aged; Ovalbumin; Polymorphism, Single Nucleotide; Signal Transduction; Th1 Cells; Toll-Like Receptor 5

The use of antibiotics during pregnancy is associated with increased allergic asthma risk in the offspring, and given that approximately 25% of pregnant women are prescribed antibiotics, it is important to understand the mechanisms contributing to this phenomenon. Currently, there are no studies that directly test this association experimentally. Our objective was to develop a mouse model in which antibiotic treatment during pregnancy results in increased offspring asthma susceptibility.. Pregnant mice were treated daily from gestation day 8-17 with an oral solution of the antibiotic vancomycin, and three concentrations were tested. At weaning, offspring were subjected to an adjuvant-free experimental asthma protocol using ovalbumin as an allergen. The composition of the gut microbiome was determined in mothers and offspring with samples collected from five different time points; short-chain fatty acids were also analyzed in allergic offspring.. We found that maternal antibiotic treatment during pregnancy was associated with increased offspring asthma severity in a dose-dependent manner. Furthermore, maternal vancomycin treatment during pregnancy caused marked changes in the gut microbiome composition in both mothers and pups at several different time points. The increased asthma severity and intestinal microbiome changes in pups were also associated with significantly decreased cecal short-chain fatty acid concentrations.. Consistent with the "Developmental Origins Hypothesis," our results confirm that exposure to antibiotics during pregnancy shapes the neonatal intestinal environment and increases offspring allergic lung inflammation.

Topics: Animals; Anti-Bacterial Agents; Asthma; Female; Humans; Hypersensitivity; Mice; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects

The aim of this study was to investigate the pharmacokinetics and pharmacodynamics of seven main active components of Mahuang decoction (MHD) and its time-concentration-effect relationship. The asthmatic rat model was established by the method of ovalbumin (OVA) sensttization. The plasma concentrations of ephedrine, pseudoephedrine, methylephedrine, amygdalin, liquiritin, cinnamic acid, glycyrrhizic acid in asthmatic model rat were investigated by a selective and rapid HPLC/MS-MS method. Simultaneously, the asthma-involved cytokines including leukotrienes B

Topics: Amygdalin; Animals; Asthma; Chromatography, High Pressure Liquid; Cinnamates; Correlation of Data; Drugs, Chinese Herbal; Ephedra sinica; Ephedrine; Flavanones; Glucosides; Glycyrrhizic Acid; Male; Ovalbumin; Plant Preparations; Rats; Rats, Sprague-Dawley

Previous studies showed a close relationship between obesity and asthma. In this study, we investigated the expression of endoplasmic reticulum (ER) stress genes in the lung tissue of obese ovalbumin (OVA)-sensitized male and female rats.. The rats were divided into eight groups (n = 5 per group) as follows: female and male rats fed with normal diet (FND and MND, respectively), female and male OVA-sensitized rats fed with normal diet (F-OND and M-OND, respectively), female and male rats fed with high-fat diet (F-HFD and M-HFD, respectively), female and male OVA-sensitized rats fed with high-fat diet (F-OHFD and M-OHFD, respectively). All rats were fed with a high-fat diet or standard pelts for 8 weeks, and for another 4 weeks, they were sensitized by OVA or saline. At the end of the study, lung tissue NF-kB protein level was assessed, and ER stress markers genes expression was determined by Real Time-PCR.. OVA-sensitization and diet-induced obesity caused the curve of methacholine concentration-response to shift to the left. In addition, the results indicated that the EC50 (the effective concentration of methacholine generating 50% of peak response) in F-OHFD rats was statistically lower than that of the M-OHFD group (p < 0.05). Moreover, the results showed that diet-induced obesity increased the expression of ATF4, ATF6, GRP78, XBP-1, and CHOP as well as the protein level of NF-kB in this experimental model of asthma, markedly in the F-OHFD group.. The results suggest that ER stress may be involved in the pathogenesis of asthma observed in obese OVA-sensitized rats, especially in the female animals.

Topics: Animals; Asthma; Diet, High-Fat; Endoplasmic Reticulum; Endoplasmic Reticulum Stress; Female; Gene Expression Regulation; HSP70 Heat-Shock Proteins; Leptin; Lung; Male; Membrane Proteins; Methacholine Chloride; NF-kappa B; Obesity; Ovalbumin; Rats; Rats, Wistar; Signal Transduction; Time Factors

Human rhinovirus (hRV) is the most common cause of asthma exacerbation characterized by clinical and pathophysiological heterogeneity. Steroid-sensitive, Th2 type-eosinophilic asthma has been somewhat studied, but hRV-induced neutrophilic asthma exacerbation is poorly understood. Here, CCR5 was found to play a role in attenuating neutrophilic airway inflammation in hRV-induced asthma exacerbation using chicken ovalbumin (OVA)-based model. CCR5 deficiency resulted in exacerbated neutrophilic asthmatic responses in airways following hRV infection. CCR5-deficient mice showed enhanced mucus expression and altered expression of tight junction proteins in lung tissues. CCR5-deficient mice were also manifested with influx of CD45

Topics: Animals; Asthma; Chemotaxis, Leukocyte; Female; Inflammation; Male; Mice, Inbred C57BL; Neutrophils; Ovalbumin; Picornaviridae Infections; Receptors, CCR5; Rhinovirus; Symptom Flare Up; T-Lymphocytes, Regulatory; Th17 Cells

N-acetylcysteine (NAC) affects signaling pathways involved in apoptosis, angiogenesis, cell growth and arrest, redox-regulated gene expression, and the inflammatory response. However, it is not known how the signal mechanism for tight junctional protein claudin (CLDN) 18 is regulated in asthma patients.. To investigate the effects of NAC on CLDN18 expression in a mouse model of asthma, and to assess plasma levels of CLDN18 in asthma patients. A murine model of asthma induced by ovalbumin (OVA) was established using wild-type BALB/c female mice, and the levels of CLDNs, phosphorylated-pyruvate dehydrogenase kinase 1 (p-PDK1), and protein kinase B (Akt) pathway proteins following NAC treatment were examined by Western blotting and immunohistochemistry. In addition, the plasma levels of CLDN18 were evaluated in asthmatic patients and control subjects.. NAC diminished OVA-induced airway hyper-responsiveness and inflammation. Levels of CLDN18 protein were higher in lung tissue from OVA mice than tissue from control mice, and were increased by treatment with NAC or dexamethasone. Treatment with NAC or dexamethasone decreased the OVA-induced increase in interleukin-1α protein levels. Although treatment with NAC increased OVA-induced p-PDK1 protein levels, it decreased phosphorylated Akt (pAkt)/Akt levels. Soluble CLDN18 levels were lower in patients with asthma than in controls and were correlated with the percentage of neutrophils, forced expiratory volume in 1 second (FEV1)/forced vital capacity % (FVC%) and FEV1%.. CLDN18 plays a role in the pathogenesis of asthma and NAC diminishes airway inflammation and responsiveness by modulating CLDN18 expression.

Topics: Acetylcysteine; Animals; Asthma; Bronchoalveolar Lavage Fluid; Claudins; Disease Models, Animal; Female; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

To investigate the effect of gestational and lactational nonylphenol (NP) exposure on airway inflammation in ovalbumin (OVA)-induced asthmatic pups. Dams were gavaged with NP at dose levels of 25 mg/kg/day (low dose), 50 mg/kg/day (middle dose), 100 mg/kg/day (high dose) and groundnut oil alone (vehicle control) respectively from gestational day 7 to postnatal day 21. The results showed that the NP content in the lung tissues of pups in the 100 mg/kg NP group was significantly higher than that of the control group (P = 0.004). In the 100 mg/kg NP group, the infiltration of lymphocytes and eosinophils with thicken smooth muscle layer and inflammatory cells in the lumen were observed in the lung tissues of pups. Osmiophilic lamellar bodies were found in the cytoplasm of type II epithelial cells; mitochondria were clearly swollen. Compared with the control group, the levels of interleukin-4 (IL-4) in BALF (P = 0.042) and ovalbumin-specific serum immunoglobulin E (OVA-sIgE) (P = 0.005) in the OVA group were significantly higher. 25 mg/kg NP-OVA co-exposure synergistically decreased nuclear factor-κB (NF-κB) mRNA expression in the lung tissues of pups; Exposure to 50 mg/kg NP combined with OVA antagonized the increased expression of high mobility group box 1 (HMGB1) mRNA in the lung tissue. The combined exposure to 50 mg/kg NP and OVA synergistically increased HMGB1 protein expression in the lung tissues. 25 mg/kg NP-OVA co-exposure antagonized the increased nuclear factor-κB (NF-κB) protein expression in the lung tissues. There was a positive correlation between NP content and HMGB1 protein expression in the lung tissue of asthmatic pups (r = 0.602, P < 0.001). In conclusion, gestational and lactational exposure to 100 mg/kg NP in maternal rats exacerbated airway inflammation in OVA-induced asthmatic pups, and there is an interactive effect between NP and OVA. When the perinatal rats were exposed to 100 mg/kg NP, the levels of HMGB1 and NF-κB in the lung tissues of OVA-induced asthmatic pups were increased.

Topics: Animals; Asthma; Disease Models, Animal; Environmental Pollutants; Female; Inflammation; Lung; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Phenols; Pregnancy; Rats; Toxicity Tests

Topics: Animals; Anti-Asthmatic Agents; Asthma; Cell Line; Child; Female; Gene Expression Regulation; Goblet Cells; Hedgehog Proteins; Humans; Interleukin-13; Interleukin-4; Interleukin-5; Janus Kinases; Mice; Mice, Inbred BALB C; Ovalbumin; Primary Cell Culture; Pyrimidines; Pyroglyphidae; Respiratory Mucosa; Signal Transduction; STAT6 Transcription Factor; Transcription, Genetic; Uteroglobin

Goat milk (GM), as compared to cow milk (CM), is easier for humans to digest. It also has antioxidant and anti-inflammatory effects and can improve minor digestive disorders and prevent allergic diseases in infants. It is unclear whether GM consumed in pregnant mothers has any protective effects on allergic diseases in infants. In this experimental study with mice, we found GM feeding enhanced immunoglobulin production, antigen-specific (ovalbumin, OVA) immune responses, and phagocytosis activity. The GM-fed mice had an increasing proportion of CD3

Topics: Adaptive Immunity; Allergens; Animals; Animals, Newborn; Asthma; Dermatophagoides pteronyssinus; Disease Models, Animal; Female; Gastrointestinal Microbiome; Goats; Immunity, Innate; Male; Mice; Mice, Inbred BALB C; Milk; Ovalbumin; Pregnancy

Bronchial asthma is a chronic disease characterized by inflammation, obstruction, and hyperresponsiveness of the airways. There is currently no curative therapy for asthma. Type 2 helper T cell response plays a critical role in the pathogenesis of the disease. Protein S is a glycoprotein endowed with anticoagulant, anti-inflammatory, and anti-apoptotic properties. Whether protein S can suppress bronchial asthma and be useful for its therapy is unknown.. To address this question here we compared the development of allergen-associated bronchial asthma between wild type and protein S-overexpressing transgenic mice. Mice were sensitized and challenged with ovalbumin. We also evaluated the circulating levels of total and active protein S in patients with bronchial asthma and healthy controls.. The circulating level of total protein S and of its active form was significantly decreased in patients with bronchial asthma compared to controls. Allergic protein S transgenic mice showed a significant reduction of airway hyperresponsiveness, lung tissue inflammatory cell infiltration, lung levels of Th2 cytokines and IgE compared to their wild-type counterparts. Administration of exogenous human protein S also decreased airway hyperresponsiveness and Th2-mediated lung inflammation in allergic wild-type mice compared with their untreated mouse counterparts. Human protein S significantly shifted the Th1/Th2 balance to Th1 and promoted the secretion of Th1 cytokines (IL-12, tumor necrosis factor-α) from dendritic cells.. These observations suggest the strong protective activity of protein S against the development of allergic bronchial asthma implicating its potential usefulness for the disease treatment.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Cytokines; Disease Models, Animal; Humans; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Protein S; Th2 Cells

Asthma is a chronic allergic respiratory disease with limited therapeutic options. Here we validated the potential anti-inflammatory, anti-asthmatic and immunomodulatory therapeutic properties of calcio-herbal ayurvedic formulation, Divya-Swasari-Ras (DSR) in-vivo, using mouse model of ovalbumin (OVA) induced allergic asthma. HPLC analysis identified the presence of various bioactive indicating molecules and ICP-OES recognized the presence of Ca mineral in the DSR formulation. Here we show that DSR treatment significantly reduced cardinal features of allergic asthma including inflammatory cell accumulation, specifically lymphocytes and eosinophils in the Broncho-Alveolar Lavage (BAL) fluids, airway inflammation, airway remodelling, and pro-inflammatory molecules expression. Conversely, number of macrophages recoverable by BAL were increased upon DSR treatment. Histology analysis of mice lungs revealed that DSR attenuates inflammatory cell infiltration in lungs and thickening of bronchial epithelium. PAS staining confirmed the decrease in OVA-induced mucus secretion at the mucosal epithelium; and trichrome staining confirmed the decrease in peribronchial collagen deposition upon DSR treatment. DSR reduced the OVA-induced pro-inflammatory cytokines (IL-6, IL-1β and TNF-α) levels in BALF and whole lung steady state mRNA levels (IL-4, -5, -33, IFN-γ, IL-6 and IL-1β). Biochemical assays for markers of oxidative stress and antioxidant defence mechanism confirmed that DSR increases the activity of SOD, Catalase, GPx, GSH, GSH/GSSG ratio and decreases the levels of MDA activity, GSSG, EPO and Nitrite levels in whole lungs. Collectively, present study suggests that, DSR effectively protects against allergic airway inflammation and possess potential therapeutic option for allergic asthma management.

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Interleukin-6; Lung; Male; Medicine, Ayurvedic; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Preparations; Tumor Necrosis Factor-alpha

The airway inflammatory response is closely associated with asthma. The purpose of this article was to study the roles of innate lymphoid cells (ILCs) in the process of airway inflammatory response in asthma. We established the asthmatic mice model with intraperitoneal injected ovalbumin medium, then with the flow cytometry analysis, we detected the ILCs and their surface proteins in the mice blood samples, besides, we analyzed the amounts of inflammatory cytokines and secreted proteins in the mice bronchoalveolar lavage fluid and blood serum. Moreover, Western blot analyzed the proteins in the mice bronchial epithelial tissues. The ILC2 amounts were obviously increased in young asthmatic mice model. And, the proteins CD25 and CCR10 were highly expressed in the sorted ILC2s. Besides, the cytokines interleukin (IL)-5, IL-13, IL-33, CCL22, and CCL27 were abundant in the bronchoalveolar lavage fluid of asthmatic mice model. And, the secretion of IL-5, IL-13, IL-33, TSLP, and CCL22 in blood serum was much more in asthmatic mice model than in the normal control mice, whereas the secretion of PGD2 was suppressed in asthmatic mice bronchoalveolar lavage fluid and blood serum. Additionally, the guanine nucleotide-binding proteins Gα12 and Gα13 were upregulated in asthmatic mice bronchial tissues, and the protein SERCA2 was downregulated; moreover, the proteins NFAT, IRF4, and its downstream signal STAT6 were all upregulated in the asthmatic mice bronchial tissues. ILC2s were involved in the response of airway inflammation through secretion of proinflammatory cytokines and chemokines to dysregulate the Ca

Topics: Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Immunity, Innate; Inflammation; Injections, Intraperitoneal; Lymphocytes; Male; Mice; Mice, Inbred BALB C; Ovalbumin

The anti-inflammatory and anti-asthmatic effects of rubusoside (Rbs) were investigated in the ovalbumin (OVA)-induced asthmatic mice, followed by effective attenuation of Rbs treatment on the airway hyperresponsiveness and reduction of inflammatory cells inside the bronchoalveolar lavage fluid (BALF). The mitigation of inflammatory infiltration as a result of Rbs treatment was histologically observed in these mice lungs. Rbs contributed to the decrease of inflammatory cytokines (TNF-α, IL-13, IL-6, IL-5, and IL-4) inside the BALF of mice with asthma. A decline of OVA-dependent IgE and IgG1 inside the serum was also noticed in these mice. Rbs was proved to enhance the mRNA level of Foxp3 inside the mice lung affected with asthma while decrease that of IL-17A, IL-23, and RORγt. NF-κB pathway activation elicited by OVA was suppressed by Rbs inside the pulmonary tissues. Rbs played significantly in the reduction of airway inflammation induced by OVA which with modulating NF-κB pathway activation. PRACTICAL APPLICATIONS: Simultaneous therapy with medicine and food is strategically significant for disease prevention and treatment in traditional Chinese medicine. Rbs is a diterpene glycoside isolated from Rubus suavissimus. The anti-inflammatory and anti-asthmatic mechanism dependent of Rbs need further study clinically. The goal of current investigation is to explore the anti-inflammatory as well as anti-asthmatic activity of Rbs in mouse models of OVA-induced experimental allergic asthma. Results of the present study are scientifically supportive for the use of Rbs as an adjunctive reagent for clinical treatment of allergic asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Diterpenes, Kaurane; Glucosides; Mice; NF-kappa B; Ovalbumin

During chronic inflammatory disease, such asthma, leukocytes can invade the central nervous system (CNS) and together with CNS-resident cells, generate excessive reactive oxygen species (ROS) production as well as disbalance in the antioxidant system, causing oxidative stress, which contributes a large part to neuroinflammation. In this sense, the aim of this study is to investigate the effects of treatment with neostigmine, known for the ability to control lung inflammation, on oxidative stress in the cerebral cortex of asthmatic mice. Female BALB/cJ mice were submitted to asthma model induced by ovalbumin (OVA). Control group received only Dulbecco's phosphate-buffered saline (DPBS). To evaluate neostigmine effects, mice received 80 μg/kg of neostigmine intraperitoneally 30 min after each OVA challenge. Our results revealed for the first time that treatment with neostigmine (an acetylcholinesterase inhibitor that no crosses the BBB) was able to revert ROS production and change anti-oxidant enzyme catalase in the cerebral cortex in asthmatic mice. These results support the communication between the peripheral immune system and the CNS and suggest that acetylcholinesterase inhibitors, such as neostigmine, should be further studied as possible therapeutic strategies for neuroprotection in asthma.

Topics: Animals; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Catalase; Cerebral Cortex; Cholinesterase Inhibitors; Female; Injections, Intraperitoneal; Mice; Mice, Inbred BALB C; Neostigmine; Neuroprotection; Neuroprotective Agents; Ovalbumin; Oxidative Stress; Reactive Oxygen Species; Sodium-Potassium-Exchanging ATPase; Superoxide Dismutase-1

Diisononyl phthalate (DINP) is commonly used as a plasticizer in industrial and consumer product applications. Several studies have suggested a possible link between exposure to DINP and the development of allergic asthma, and the synergistic effect of DINP combined with Ovalbumin (OVA) is a possible way to promote an immune response. These findings are still speculative, since there is insufficient evidence to assess the ability of DINP to influence "allergic asthma pathology". This study was designed to determine any effects of OVA/DINP exposure on airway reactivity, particularly when combined with allergen exposure. Experiments to determine these effects were conducted after 15 days of combined exposure and a subsequent challenge with aerosolized ovalbumin for one week. Airway hyper-responsiveness (lung function), lung tissue pathology, cytokines and oxidative stress biomarkers were investigated. We showed that oral exposure to OVA/DINP could induce airway hyper-responsiveness (AHR), and aggravate airway wall remodeling, and that this deterioration was concomitant with increased immunoglobulin-E and Th2 cytokines secretion. The data also demonstrated that DINP could promote oxidative damage in the lung. In summary, this study showed that DINP has an adjuvant effect on allergic asthma affecting lung function, lung histopathology, immune molecules and causes oxidative damage.

Topics: Adjuvants, Immunologic; Animals; Asthma; Biomarkers; Mice; Ovalbumin; Oxidative Stress; Phthalic Acids; Respiratory Hypersensitivity

This study was designed to investigate the effects of low concentration hydrogen inhalation on asthma and sleep function in mice and the potential mechanism.. In the asthma experiment, BALB/c mice were randomly divided into normal control group, asthma model group and hydrogen treatment group. After establishing ovalbumin (OVA)-induced asthma model, the hydrogen treatment group mice were treated by inhalation of hydrogen (24-26 mL/L per day) for 7 consecutive days, and the normal control group and asthma model group mice received similar treatment by inhalation of air. The levels of interleukin (IL)-4, IL-13, and interferon-γ (IFN-γ) in bronchoalveolar lavage fluid (BALF) were measured by commercially available ELISA kits. The levels of malondialdehyde (MDA) and glutathione (GSH), as well as the activity of superoxide dismutase (SOD) in lung tissue were detected by colorimetric assays. The pathological changes in lung tissue were assessed by HE staining. In the sleep experiment, ICR mice were randomly divided into blank control group and 1 d, 3 d, 5 d hydrogen treatment groups and diazepam group. The effects of inhalation of 24-26 mL/L per day hydrogen on the sleep duration induced by intraperitoneal injection of upper-threshold dose of sodium pentobarbital and the sleep latency in response to subthreshold dose were evaluated.. In the asthma experiment, the asthma model group showed higher levels of IL-4 and IL-13 (. Low concentration hydrogen inhalation could alleviate OVA-induced asthma in mice, and the mechanism might be related to the anti-oxidative and anti-inflammatory effects of hydrogen. Also, low concentration hydrogen inhalation could improve sleep function in mice.

Topics: Administration, Inhalation; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Hydrogen; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred ICR; Ovalbumin; Sleep

Some studies have shown that maturation of dendritic cells (DCs) is modulated directly by pathogen components via pattern recognition receptors such as Toll-like receptors, but also by signal like CD40 ligand (CD40 L or CD154) mediated by activated T cells. Several reports indicate that invariant natural killer T (iNKT) cells up-regulate CD40 L upon stimulation and thereby induce activation and maturation of DCs through crosslink with CD40. Our previous findings indicated that iNKT cells promote Th2 cell responses through the induction of immunogenic maturation of lung DCs (LDCs) in the asthmatic murine, but its mechanism remains unclear. Therefore, we investigated the immunomodulatory effects of blockade of CD40 L using anti-CD40 L treatment on Th2 cell responses and immunogenic maturation of LDCs, and further analyzed whether these influences of blockade of CD40 L were related to lung iNKT cells using iNKT cell-deficient mice and the combination treatment of specific iNKT cell activation with anti-CD40 L treatment in murine models of asthma. Our findings showed that blockade of CD40 L using anti-CD40 L treatment attenuated Th2 cell responses in wild-type (WT) mice, but not in CD1d-deficient mice sensitized and challenged with ovalbumin (OVA) or house dust mite (HDM). Meanwhile, blockade of CD40 L down-regulated immunogenic maturation of LDCs in WT mice, but not in CD1d-deficient mice sensitized and challenged with OVA. Additionally, agonistic anti-CD40 treatment reversed the inhibitory effects of anti-CD40 L treatment on Th2 cell responses and LDC activation in an OVA-induced mouse model of asthma. Furthermore, LDCs from asthmatic mice treated with anti-CD40 L could significantly reduce the influence on Th2 cell responses in vivo and in vitro. Finally, α-Galactosylceramide plus anti-CD40 L treatment stimulated lung iNKT cells, but suppressed Th2 cell responses in the asthmatic mice. Taken together, our data raise an evidence that blockade of CD40 L attenuates Th2 cell responses through the inhibition of immunogenic maturation of LDCs, which may be at least partially related to lung iNKT cells in murine models of asthma.

Topics: Animals; Antigens, CD1d; Asthma; CD40 Ligand; Cell Communication; Dendritic Cells; Disease Models, Animal; Female; Galactosylceramides; Humans; Lung; Mice; Mice, Knockout; Natural Killer T-Cells; Ovalbumin; Pyroglyphidae; Th2 Cells

Airway remodeling happens in childhood asthma, in parallel with, but not necessarily subsequent to, airway inflammation. The differentiation of airway epithelial cells into myofibroblasts via epithelial-mesenchymal-transition (EMT) is one of the mechanisms underlying airway remodeling. This study aimed at identifying novel molecules involved in pediatric asthma-associated airway remodeling. Asthma model was established by challenging C57BL/6 mouse pups with ovalbumin (OVA). We found that the expression of Cadherin 11 (CDH11), a type II cadherin, was increased by OVA treatments in the airway epithelium. Our earlier microarray data suggested miRNA-451a-5p (miRNA-451a) as a potential regulator of CDH11. In contrast to CDH11, miRNA-451a expression decreased in the asthmatic lung. MiRNA-451a was then packaged into a lentivirus vector and systematically given to the asthmatic pups. Our data indicated that OVA-induced infiltration of inflammatory cells, including eosnophils, neutrophils, macrophages and lymphocytes, was reduced by miRNA-451a over-expression. EMT was initiated in asthmatic mice as demonstrated by increased alpha-smooth muscle actin (α-SMA) positive cells present in airway epithelium, which was inhibited by miRNA-451a. CDH11 elevation in vivo was also inhibited by miRNA-451a. Dual-Luciferase analysis further showed CDH11 as a novel valid target of miRNA-451a. Additionally, in vitro, EMT was triggered in human 16HBE airway epithelial cells by pro-fibrotic transforming growth factor β (TGF-β). Corresponding to the anti-EMT effects observed in vivo, miRNA-451a also inhibited TGF-β-induced collagen deposition in cultured airway epithelial cells by targeting in CDH11. In summary, our study demonstrates that the deregulated miRNA-451a-CDH11 axis contributes to airway remodeling in childhood asthma.

Topics: Airway Remodeling; Allergens; Animals; Animals, Newborn; Asthma; Cadherins; Cells, Cultured; Disease Models, Animal; Humans; Hypersensitivity; Male; Mice; Mice, Inbred C57BL; MicroRNAs; Ovalbumin; Respiratory Mucosa; Signal Transduction; Transforming Growth Factor beta

What factors and underlying mechanisms influence the occurrence of the atopic march remain unclear. Recent studies suggest that exposure to diisononyl phthalate (DINP) might be associated with the occurrence of atopic dermatitis (AD) and asthma. However, little is known about the role of DINP exposure in the atopic march. In this study, we investigated the effect of DINP exposure on the progression from AD to asthma, and explored the potential mechanisms. We built an atopic march mouse model from AD to asthma, by exposure to DINP and sensitization with OVA. Pyrrolidine dithiocarbamate and SB203580 were used to block NF-κB and p38 MAPK respectively, to explore the possible molecular mechanisms. The data showed that DINP aggravated airway remodeling and airway hyperresponsiveness (AhR) in the progression from AD to asthma, induced a sharp increase in IL-33, IgE, Th2 and Th17 cytokines, and resulted in an increase in the expression of thymic stromal lymphopoietin (TSLP) and in the number of inflammatory cells. Blocking NF-κB inhibited AD-like lesions, and the production of IL-33 and TSLP in the progression of AD, while alleviating airway remodeling, AhR, and the expression of Th2 and Th17 cytokines in both the progression of AD and the asthmatic phenotype. Blocking p38 MAPK in the progression of asthma, inhibited airway remodeling, AhR, and the expression of Th2 and Th17 cytokines. The results demonstrated that exposure to DINP enhanced the immune response to memory CD4

Topics: Airway Remodeling; Animals; Asthma; Cytokines; Dermatitis, Atopic; Disease Models, Animal; Disease Progression; Enzyme Activation; Hypersensitivity, Immediate; Male; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Phthalic Acids; Respiratory Hypersensitivity; Signal Transduction; Specific Pathogen-Free Organisms; Th17 Cells; Th2 Cells; Thymic Stromal Lymphopoietin

BACKGROUND Bifidobacteria are among the probiotics used in treating intestinal diseases and are rarely used for allergic asthma treatment. The present study investigated the mechanism of B. infantis in treating allergic asthma in mice. MATERIAL AND METHODS A total of 40 male Balb/c mice were randomized into control, ovalbumin (OVA), montelukast (Mon), and B. infantis (B10) groups, and allergic asthma was induced in the OVA, Mon, and B10 groups. Airway reactivity was measured on day 29 by methacholine at various doses. The numbers of total cells and inflammatory cells in bronchoalveolar lavage fluid (BALF) were counted by blood cell counter and Diff-Quik staining. Hematoxylin-eosin (HE) staining was performed to observe inflammatory cell infiltration in lung tissues. Total IgE and OVA-specific IgE in serum were measured by ELISA. Mucin 5AC expression was detected by Western blot to evaluate airway obstruction. The levels of Th1 (IFN-γ, IL-2) and Th2 (IL-4, IL-5, IL-13) cytokines in BALF and tissues were detected by ELISA and qRT-PCR, respectively. RESULTS The mice in the OVA group had airway hyperreactivity, while the symptoms in the B10 group and Mon group were effectively relieved. B10 reduced the number of inflammatory cells in BALF as well as inflammatory cell infiltration in tissues. Moreover, the levels of total serum IgE, OVA-specific IgE, and Mucin 5AC were increased in the OVA group, but were reduced in the Mon group and B10 group. B. infantis increased the levels of Th1 cytokines and decreased those of Th2 cytokines. CONCLUSIONS B. infantis can reduce the infiltration of inflammatory cells induced by OVA-specific antibodies in mice. B. infantis has therapeutic effects on allergic asthma by promoting Th1 and inhibiting Th2 immune responses.

Topics: Animals; Asthma; Bifidobacterium longum subspecies infantis; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Random Allocation; Th1 Cells; Th2 Cells

Peroxisome proliferator-activated receptor gamma (PPARγ) is a multifaceted ligand-activated transcription factor that regulates inflammatory responses in asthma pathophysiology. The present study corroborates PPARγ-mediated anti-asthmatic action of the flavonoid, galangin (norizalpinin). In silico molecular interactions reveal that galangin formed three H-bonds (Glu291, Leu340 and Ser342) and a π-sigma bond (Arg288) with PPARγ, contributing to the binding affinity and stability of the complex. In vivo studies explore the role of galangin as a propitious PPARγ agonist in mitigating airway inflammation, thereby excluding ligand-independent action of PPARγ. Accordingly, oral administration of galangin significantly ameliorated airway hyperresponsiveness, inflammation and goblet cell hyperplasia by the suppression of IL-4, 5, 13, 17, TNF-α, NO, ROS, EPO, IgE and increase of IFN-γ in ovalbumin-induced allergic asthma model. PPARγ expression (mRNA and protein) studies were performed to elucidate a possible mechanism by which galangin modulates. Furthermore, to eliminate PPARγ-independent effects of galangin, a specific PPARγ antagonist (GW9662) was administered, which dramatically reversed the effects of galangin on PPARγ up-regulation, confirming the pleiotropic role of galangin as a PPARγ agonist in asthma therapeutics. Taken together, our findings communicate that PPARγ plays as a master regulator in the anti-asthmatic action of galangin.

Topics: Amino Acid Sequence; Anilides; Animals; Anti-Asthmatic Agents; Asthma; Binding Sites; Biomechanical Phenomena; Female; Flavonoids; Gene Expression Regulation; Humans; Hydrogen Bonding; Interleukins; Lung; Mice, Inbred BALB C; Molecular Docking Simulation; Ovalbumin; PPAR gamma; Protein Binding; Protein Conformation; RNA, Messenger; Signal Transduction; Tumor Necrosis Factor-alpha

Parasympathetic neurons in the airways control bronchomotor tone. Increased activity of cholinergic neurons are mediators of airway hyperresponsiveness (AHR) in asthma, however, mechanisms are not elucidated. We describe remodeling of the cholinergic neuronal network in asthmatic airways driven by brain-derived neurotrophic factor (BDNF) and Tropomyosin receptor kinase B (TrkB). Human bronchial biopsies were stained for cholinergic marker vesicular acetylcholine transporter (VAChT). Human lung gene expression and single nucleotide polymorphisms (SNP) in neuroplasticity-related genes were compared between asthma and healthy patients. Wild-type (WT) and mutated TrkB knock-in mice (Ntrk2tm1Ddg/J) with impaired BDNF signaling were chronically exposed to ovalbumin (OVA). Neuronal VAChT staining and airway narrowing in response to electrical field stimulation in precision cut lung slices (PCLS) were assessed. Increased cholinergic fibers in asthmatic airway biopsies was found, paralleled by increased TrkB gene expression in human lung tissue, and SNPs in the NTRK2 [TrkB] and BDNF genes linked to asthma. Chronic allergen exposure in mice resulted in increased density of cholinergic nerves, which was prevented by inhibiting TrkB. Increased nerve density resulted in AHR in vivo and in increased nerve-dependent airway reactivity in lung slices mediated via TrkB. These findings show cholinergic neuroplasticity in asthma driven by TrkB signaling and suggest that the BDNF-TrkB pathway may be a potential target.

Topics: Adolescent; Animals; Asthma; Case-Control Studies; Cholinergic Agents; Female; Humans; Inflammation; Lung; Male; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Muscle, Smooth; Neuronal Plasticity; Ovalbumin; Polymorphism, Single Nucleotide; Receptor, trkB; Signal Transduction

The current study aimed to study the effects of Bulleyaconitine A (BLA) on asthma. Asthmatic mice model was established by ovalbumin (OVA) stimulation, and the model mice were treated by BLA. After BLA treatment, the changes in lung and airway resistances, total and differential leukocytes in the bronchoalveolar lavage fluid (BALF) were detected, and the changes in lung inflammation and airway remodeling were observed. Moreover, the secretion of IgE, Th1/Th2-type and IL-17A cytokines in BALF and serum of the asthmatic mice were determined. The resuts showed that BLA attenuated OVA-induced lung and airway resistances, inhibited the inflammatory cell recruitment in BALF and the inflammation and airway remodeling of the asthmatic mice. In addition, BLA suppressed the secretion of IgE, Th2-type cytokines, and IL-17A, but enhanced secretions of Th1-type cytokines in BALF and serum. The current study discovered that BLA inhibited the lung inflammation and airway remodeling via restoring the Th1/Th2 balance in asthmatic mice.

Topics: Aconitine; Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Immunoglobulin E; Interleukin-17; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Signal Transduction; Th1 Cells; Th1-Th2 Balance; Th2 Cells; Treatment Outcome

Topics: Allergens; Animals; Asthma; Eosinophilia; Heme Oxygenase-1; Membrane Proteins; Mice, Inbred BALB C; Ovalbumin; Phosphorylation; Signal Transduction; STAT3 Transcription Factor; Suppressor of Cytokine Signaling 3 Protein; Th17 Cells; Th2 Cells

Prenatal challenges such as maternal stress perception increase the risk and severity of asthma during childhood. However, insights into the trajectories and targets underlying the pathogenesis of prenatally triggered asthma are largely unknown. The developing lung and immune system may constitute such targets.. Here we have aimed to identify the differential sex-specific effects of prenatal challenges on lung function, immune response, and asthma severity in mice.. We generated bone marrow chimeric (BMC) mice harboring either prenatally stress-exposed lungs or a prenatally stress-exposed immune (hematopoietic) system and induced allergic asthma via ovalbumin. Next-generation sequencing (RNA sequencing) of lungs and assessment of airway epithelial barrier function in ovalbumin-sensitized control and prenatally stressed offspring was also performed.. Profoundly enhanced airway hyperresponsiveness, inflammation, and fibrosis were exclusively present in female BMC mice with prenatally stress-exposed lungs. These effects were significantly perpetuated if both the lungs and the immune system had been exposed to prenatal stress. A prenatally stress-exposed immune system alone did not suffice to increase the severity of these asthma features. RNA sequencing analysis of lungs from prenatally stressed, non-BMC, ovalbumin-sensitized females unveiled a deregulated expression of genes involved in asthma pathogenesis, tissue remodeling, and tight junction formation. It was also possible to independently confirm a tight junction disruption. In line with this, we identified an altered perinatal and/or postnatal expression of genes involved in lung development along with an impaired alveolarization in female prenatally stressed mice.. Here we have shown that the fetal origin of asthma is orchestrated by a disrupted airway epithelium and further perpetuated by a predisposed immune system.

Topics: Animals; Asthma; Bone Marrow; Cells, Cultured; Disease Models, Animal; Female; Immunity; Inflammation; Lung; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; Respiratory Hypersensitivity; Respiratory Mucosa; Tight Junctions

Asthma is a heterogeneous disease characterized by chronic inflammation and structural changes in the airways. The airway smooth muscle (ASM) is responsible for airway narrowing and an important source of inflammatory mediators. We and others have previously shown that WNT5A mRNA and protein expression is higher in the ASM of asthmatics compared to healthy controls. Here, we aimed to characterize the functional role of (smooth muscle-derived) WNT5A in asthma. We generated a tet-ON smooth-muscle-specific WNT5A transgenic mouse model, enabling in vivo characterization of smooth-muscle-derived WNT5A in response to ovalbumin. Smooth muscle specific WNT5A overexpression showed a clear trend towards enhanced actin (α-SMA) expression in the ASM in ovalbumin challenged animals, but had no effect on collagen content. WNT5A overexpression in ASM also significantly enhanced the production of the Th2-cytokines IL4 and IL5 in lung tissue after ovalbumin exposure. In line with this, WNT5A increased mucus production, and enhanced eosinophilic infiltration and serum IgE production in ovalbumin-treated animals. In addition, CD4

Topics: Actins; Airway Remodeling; Allergens; Animals; Asthma; CD4-Positive T-Lymphocytes; Cell Movement; Eosinophils; Female; Gene Expression Regulation; Humans; Immunoglobulin E; Interleukin-4; Interleukin-5; Interleukins; Lung; Lymphocyte Activation; Mice; Mice, Transgenic; Muscle, Smooth; Ovalbumin; Primary Cell Culture; Transgenes; Wnt-5a Protein

To observe the effects of artesunate on eosinophil (EOS) apoptosis and Fas and Bcl-2 protein expressions in asthmatic mice.. Thirty female BALB/c mice aged 6-8 weeks were randomly divided into control group, asthma group and artesunate group. Except for those in the control group, all the mice were sensitized with aerosolized ovalbumin to establish mouse models of asthma. In artesunate group, the rats were intraperitoneally injected with artesunate 1 h before ovalbumin inhalation from the 21st day of modeling. The lung tissues were harvested for staining 24 h after the last challenge. Flow cytometry was used to analyze the percentage and apoptosis rate of EOS in the alveolar lavage fluid (BALF). The apoptosis of EOS in the lung tissue was detected with TUNEL method, and Fas and Bcl-2 protein expressions were detected using immunohistochemistry.. Compared with those in asthma group, the artesunate-treated mice had significantly decreased percentage of EOS in the BALF (. Artesunate regulates the protein expressions of Fas and Bcl-2 to reduce EOS infiltration in the lung tissue and promote EOS apoptosis in asthmatic mice.

Topics: Animals; Apoptosis; Artesunate; Asthma; Bronchoalveolar Lavage Fluid; Eosinophils; fas Receptor; Female; Mice; Mice, Inbred BALB C; Ovalbumin; Proto-Oncogene Proteins c-bcl-2; Random Allocation

In this study, a mice model of obesity-asthma was established. We investigated the correlation between oxidative stress and NF-κB signaling pathway in the lung tissues, together with the effects of acetylcysteine. The animals were fed on a high-fat diet, and then ovalbumin (OVA) sensitization was utilized to establish the obesity-asthma model. N-acetylcysteine was used to treat asthma, animals treated with budesonide served as control. The malondialdehyde (MDA) in the lung tissues was determined, together with the activity of glutathione (GSH). EMAS assay was utilized to measure the nuclear factor-κB-P65 (NF-κB-P65) in lung tissues. Western blot analysis was performed to determine the expression of inhibitor kappa B-α (IκB-α) and inhibitor kappa B kinase-β (IKK-β). The MDA in the asthma groups showed significantly elevation (P < 0.01), and the GSH showed significant decrease (P < 0.01), especially in the obesity-asthma group. The efficiency of N-acetylcysteine was superior to that of the budesonide in the decline of MDA and elevation of GSH (P < 0.01). In both asthma groups, the expression of IKK-β and transcription of NF-κB-P65 in the lung tissues showed significant elevation (P < 0.01), and IκB-α showed significant decline (P < 0.01), especially in the obesity-asthma group. There was decline of IKK-β and NF-κB-P65 and elevation of IκB-α in the N-acetylcysteine group, which was even significantly in the Budesonide group (P < 0.01). There was a positive correlation between MDA and NF-κB activation in the lung tissues in all the asthma groups and treatment groups (P < 0.05). Obesity-asthma mice showed higher oxidative stress and activation of NF-κB compared with that of the asthma mice. There was a positive correlation between MDA and NF-κB activation in the lung tissues in the asthma groups. N-acetylcysteine was more effective in reducing the oxidative stress compared to the budesonide.

Topics: Acetylcysteine; Animals; Asthma; Female; Glutathione; I-kappa B Kinase; Lung; Malondialdehyde; Mice; Mice, Inbred C57BL; Mice, Obese; NF-kappa B; NF-KappaB Inhibitor alpha; Obesity; Ovalbumin; Oxidative Stress; Phosphorylation; Signal Transduction; Tumor Necrosis Factor-alpha

Allergic asthma is the most common phenotype of the pathology, having an early-onset in childhood and producing a Th2-driven airways remodeling process that leads to symptoms and pathophysiological changes. The avoidance of aeroallergen exposure in early life has been shown to prevent asthma, but without repeated success and with the underlying preventive mechanisms at the beginning of asthma far to be fully recognized. In the present study, we aimed to evaluate if neonatal LPS-induced boost in epithelial host defenses contribute to prevent OVA-induced asthma in adult mice. To this, we focused on the response of bronchiolar club cells (CC), which are highly specialized in maintaining the epithelial homeostasis in the lung. In these cells, neonatal LPS administration increased the expression of TLR4 and TNFα, as well as the immunodulatory/antiallergic proteins: club cell secretory protein (CCSP) and surfactant protein D (SP-D). LPS also prevented mucous metaplasia of club cells and reduced the epidermal growth factor receptor (EGFR)-dependent mucin overproduction, with mice displaying normal breathing patterns after OVA challenge. Furthermore, the overexpression of the epithelial Th2-related molecule TSLP was blunted, and normal TSLP and IL-4 levels were found in the bronchoalveolar lavage. A lower eosinophilia was detected in LPS-pretreated mice, along with an increase in phagocytes and regulatory cells (CD4+CD25+FOXP3+ and CD4+IL-10+), together with higher levels of IL-12 and TNFα. In conclusion, our study demonstrates stable asthma-preventive epithelial effects promoted by neonatal LPS stimulation, leading to the presence of regulatory cells in the lung. These anti-allergic dynamic mechanisms would be overlaid in the epithelium, favored by an adequate epidemiological environment, during the development of asthma.

Topics: Allergens; Animals; Animals, Newborn; Asthma; Bronchioles; Cytokines; Disease Models, Animal; Epithelium; Immunity, Innate; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory

Objective To investigate whether the recombinant pyrin domain protein can alleviate the airway inflammation and airway remodeling of OVA-induced mice with chronic asthma by inhibiting transforming growth factor β1(TGF-β1)/SMAD and Jagged1/Notch1 signaling pathways. Methods Thirty-two male BALB/c mice were selected and divided into 4 groups with 8 mice in each group. The four groups were the control group, OVA model group, recombinant pyrin domain protein treatment group (100 μg/kg), and the dexamethasone treatment group (1 mg/kg). Enzyme-linked immunosorbent assay (ELISA) was used to determine the expression of inflammatory factors in bronchoalveolar lavage fluid (BALF) of mice in each group. hematoxylin-eosin staining (HE) was used to observe the inflammatory infiltration of bronchus in mice. The changes of goblet cells were observed by periodic acid-Schiff (PAS) staining and collagen fibers by Masson staining. Immunohistochemical staining (IHC) was performed to observe the expression distribution of α smooth muscle actin (α-SMA), TGF-β1 and Notch1 proteins in lung tissues. Western blotting was used to detect the protein levels of α-SMA, E-cadherin, TGF-β1, SMAD2/3, SMAD7, Jagged1, Notch1 and Hes1 in lung tissues. Results The recombinant pyrin domain protein not only improved the airway inflammatory response of the OVA-induced mice with bronchial asthma, but also inhibited the hyperplasia of goblet cells and collagen fiber deposition, reduced the tumor necrosis factor α (TNF-α) in BALF, interleukin 1β (IL-1β), IL-4, IL-13 levels, and inhibited the protein expression of TGF-β1, SMAD2/3, Jagged1, Notch1, Hes1 and α-SMA in lung tissues. Conclusion The recombinant pyrin domain protein can reduce the airway inflammation and airway remodeling of asthmatic mice by inhibiting TGF-β1/SMAD and Jagged1/Notch1 signaling pathways.

Topics: Airway Remodeling; Animals; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Jagged-1 Protein; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Pyrin Domain; Receptor, Notch1; Recombinant Proteins; Signal Transduction; Smad Proteins; Transforming Growth Factor beta1

Androgen levels inversely correlate with the incidence, susceptibility and severity of asthma. However, whether male sex hormones such as 5α-dihydrotestosterone (DHT) have beneficial effects on asthma symptoms and/or could affect asthma susceptibility have not been investigated. DHT administration to female mice, during the sensitization phase, abrogates the sex bias in bronchial hyperreactivity. This effect correlates with inhibition of leukotriene biosynthesis in the lung. DHT significantly inhibits also other asthma-like features such as airway hyperplasia and mucus production in sensitized female mice. Conversely, DHT does not affect plasma IgE levels as well as CD3

Topics: Androgens; Animals; Asthma; Bronchial Hyperreactivity; Cell Line; Dihydrotestosterone; Female; Immunologic Factors; Male; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Sex Characteristics

The IL-33-ST2 pathway is an important initiator of type 2 immune responses. We previously characterised the HpARI protein secreted by the model intestinal nematode

Topics: Alternaria; Animals; Antigens, Helminth; Asthma; Cell Line; Humans; Immunologic Factors; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Nematospiroides dubius; Ovalbumin

Dibutyl phthalate (DBP) is one of the most ubiquitous phthalate esters found in everyday products, and is receiving increased attention as an immunologic adjuvant. However, information regarding DBP-aggravated allergic asthma is still limited. This study used a mouse model sensitized with ovalbumin (OVA) to determine any adverse effects of DBP on allergic asthma. Our results reveal that allergic asthmatic mice exposed to DBP for an extended period had a significant increase in inflammatory cell infiltration; a significant increase in levels of serum immunoglobulin and T helper 2 cell (Th2) and T helper 17 cell (Th17) cytokines in lung tissue; and significant changes in lung histology and AHR, all of which are typical asthmatic symptoms. The levels of oxidative stress and levels of the neuropeptide, calcitonin gene related peptide (CGRP), were also elevated after DBP exposure. Interestingly, blocking oxidative stress by administering melatonin (MT) not only reduced oxidative stress and CGRP levels, but also ameliorated the asthmatic symptoms. Collectively, these results show that DBP exacerbates asthma-like pathologies by increasing the expression of CGRP mediated by oxidative stress.

Topics: Animals; Asthma; Calcitonin Gene-Related Peptide; Cytokines; Dibutyl Phthalate; Disease Models, Animal; Environmental Pollutants; Lung; Melatonin; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Th17 Cells; Th2 Cells

CARAS is an airway inflammation of allergic individuals, with a type 2 immune response. The pharmacotherapy is based on drugs with relevant side effects. Thus, the goal of this study evaluated the alkaloids warifteine (War) and methylwarifteine (Mwar) from Cissampelos sympodialis in CARAS experimental model. Therefore, BALB/c mice were ovalbumin (OVA) sensitized and challenged and treated with both alkaloids. Treated animals showed a decrease (p < 0.05) of allergic signs as sneezing and nasal rubbings, histamine nasal hyperreactivity, and inflammatory cell migration into the nasal (NALF) and the bronchoalveolar (BALF) fluids, main eosinophils. In the systemic context, only Mwar reduced eosinophilia, however, both alkaloids reduced the serum levels of OVA-specific IgE. Histological analysis revealed that the alkaloids decreased the inflammatory cells into the subepithelial and perivascular regions of nasal tissue and the peribronchiolar and perivascular regions of lung tissue. Hyperplasia/hypertrophy of nasal and lung goblet cells were reduced in alkaloid treated animals; however, the treatment did not change the number of mast cells. The lung hyperactivity was attenuated by reducing hyperplasia of fibroblast and collagen fiber deposition and hypertrophy of the lung smooth muscle layer. The immunomodulatory effect was by decreasing of type 2 and 3 cytokines (IL-4/IL-13/IL-5 and IL-17A) dependent by the increasing of type 1 cytokine (IFN-γ) into the BALF of treated sick animals. Indeed, both alkaloids reduced the NF-кB (p65) activation on granulocytes and lymphocytes, indicating that the alkaloids shut down the intracellular transduction signals underlie the transcription of T

Topics: Alkaloids; Animals; Anti-Allergic Agents; Asthma; Behavior, Animal; Bronchoalveolar Lavage Fluid; Cissampelos; Collagen; Cytokines; Disease Models, Animal; Eosinophils; Female; Immunoglobulin E; Inflammation; Lung; Mast Cells; Mice, Inbred BALB C; Mucus; Nasal Lavage Fluid; Nasal Mucosa; Ovalbumin; Rhinitis, Allergic; Signal Transduction; Sneezing; Transcription Factor RelA

Neutrophilic asthma (NA) is characterized by neutrophil‑mediated inflammation and the presence of Th17 cells. However, the mechanisms underlying Th17 cell responses in NA remain unknown. The aim of the present study was to examine the effects of interleukin (IL)‑7 on Th17 cell responses in NA. A NA mouse model was sensitized by airway delivery of ovalbumin (OVA) and lipopolysaccharide and challenged with 1% OVA aerosol from day 21 for 3 consecutive days. Airway resistance was then measured to assess airway hyper‑responsiveness (AHR). Cells from bronchoalveolar lavage fluid (BALF) underwent Diff‑Quick and hematoxylin and eosin staining for classification. The levels of IL‑17 in the BALF were determined by ELISA. The effects of IL‑7 administration and STAT5 inhibition on Th17 cells were also characterized in vitro using splenic CD4+ T cells. Ki‑67, Bcl‑2 and activated caspase‑3 expression in differentiated Th17 cells were analyzed by flow cytometry. The mouse model of NA was characterized by increased AHR, elevated levels of IL‑17, high neutrophil counts in BALF, accumulated inflammatory cells in the lung and Th17 cell responses. IL‑7 promoted the expression of Ki‑67 and Bcl‑2 while reducing caspase‑3 expression. STAT5 inhibitor treatment decreased the levels of Ki‑67 and Bcl‑2, and resulted in increased expression of caspase‑3. These results suggested that the IL‑7/JAK/STAT5 signaling pathway may be involved in Th17 cell responses in NA.

Topics: Administration, Inhalation; Airway Resistance; Animals; Asthma; Bronchoalveolar Lavage Fluid; Caspase 3; Disease Models, Animal; Female; Interleukin-7; Janus Kinases; Ki-67 Antigen; Lipopolysaccharides; Lung; Mice; Mice, Inbred C57BL; Neutrophils; Ovalbumin; Proto-Oncogene Proteins c-bcl-2; Signal Transduction; STAT5 Transcription Factor; Th17 Cells

Allergic asthma is one of the most common chronic diseases of the airways, however it still remains underdiagnosed and hence undertreated. Therefore, an allergic asthma rat model would be useful to be applied in future therapeutic strategy studies. The aim of the present study was to develop an objective model of allergic asthma in atopic rats that allows the induction and quantification of anaphylactic shock with quantitative variables. Female Brown Norway rats were intraperitoneally sensitized with ovalbumin (OVA), alum and

Topics: Administration, Intranasal; Allergens; Animals; Asthma; Body Temperature; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Epithelial Cells; Female; Hypersensitivity; Hypersensitivity, Immediate; Immunoglobulin E; Leukotrienes; Lung; Ovalbumin; Rats

Tris(2-butoxyethyl) phosphate (TBEP) is a major organophosphorus flame retardant and has been widely increasing as a substitute for brominated flame retardants. TBEP may have adverse effects on human health; however, its impact on immune and allergic responses remains largely uncharacterized. In this study, the effects of low-dose TBEP comparable with the level of actual human exposure to that of human tolerable daily intake on allergic asthmatic mice were explored. Five-week-old C3H/HeJSlc male mice consumed a diet containing approximately 0.02, 0.2 or 2 μg/kg/day TBEP and were intratracheally administrated ovalbumin (OVA) (1 μg/mouse every 2 weeks from 5 to 11 weeks of age). Exposure to 2 μg/kg/day TBEP with OVA tended to enhance allergic pulmonary inflammation and significantly elevated mRNA levels of interleukin-5, eotaxin-1 and estrogen receptor alpha (ERα) compared with OVA alone. In mediastinal lymph nodes (MLNs), TBEP (0.2 or 2 μg/kg/day) with OVA significantly increased in total cell number and promoted conventional dendritic cell activation than OVA alone; MLN cell proliferation by OVA restimulation was also enhanced in these groups. In the bone marrow (BM), TBEP (0.02 or 0.2 μg/kg/day) with OVA resulted in a net decrease in total cell number and fraction of CCR2

Topics: Administration, Oral; Animals; Asthma; Bone Marrow; Cell Proliferation; Cells, Cultured; Cellular Microenvironment; Cytokines; Disease Models, Animal; Flame Retardants; Immunoglobulin E; Immunoglobulin G; Lung; Lymph Nodes; Male; Mice, Inbred C3H; No-Observed-Adverse-Effect Level; Organophosphorus Compounds; Ovalbumin; Phenotype; Th2 Cells

Asthma is characterized by inflammation, pulmonary remodeling and bronchial hyperresponsiveness. We have previously shown that treatment with angiotensin-(1-7) [Ang-(1-7)] promotes resolution of eosinophilic inflammation and prevents chronic allergic lung inflammation. Here, we evaluated the effect of treatment with the inclusion compound of Ang-(1-7) in hydroxypropyl β-cyclodextrin (HPβCD) given by inhalation on pulmonary remodeling in an ovalbumin (OVA)-induced chronic allergic lung inflammation. Mice were sensitized to ovalbumin (OVA; 4 injections over 42 days, 14 days apart) and were challenged 3 times per week, for 4 weeks (days 21-46). After the 2nd week of challenge, mice were treated with Ang-(1-7) by inhalation (4.5 μg of Ang-(1-7) included in 6.9 μg of HPβCD for 14 days, i.e. days 35-48). Mice were killed 72 h after the last challenge and blood, bronchoalveolar lavage fluid (BALF) and lungs were collected. Histology and morphometric analysis were performed in the lung. Metalloproteinase (MMP)-9 and MMP-12 expression and activity, IL-5, CCL11 in the lung and plasma IgE were measured. After 2 weeks of OVA challenge there was an increase in plasma IgE and in inflammatory cells infiltration in the lung of asthmatic mice. Treatment with inhaled administration of Ang-(1-7)/HPβCD for 14 days reduced eosinophils, IL5, CCL11 in the lung and plasma IgE. Treatment of asthmatic mice with Ang-(1-7)/HPβCD by inhalation reversed pulmonary remodeling by reducing collagen deposition and MMP-9 and MMP-12 expression and activity. These results show for the first time that treatment by inhalation with Ang-(1-7) can reverse an installed asthma, inhibiting pulmonary inflammation and remodeling.

Topics: Administration, Inhalation; Airway Remodeling; Angiotensin I; Animals; Asthma; Biomarkers; Cytokines; Disease Models, Animal; Immunoglobulin E; Lung; Matrix Metalloproteinases; Mice; Ovalbumin; Peptide Fragments; Vasodilator Agents

Topics: Animals; Anti-Asthmatic Agents; Antioxidants; Asthma; Cinnamates; Depsides; Dose-Response Relationship, Drug; Female; Hydrogen Peroxide; Lung; Mice; Mice, Inbred BALB C; NADPH Oxidases; Ovalbumin; Oxidative Stress; Rosmarinic Acid

Studies in experimental models of allergic asthma have shown that mesenchymal stem cells (MSCs) have therapeutic potential for T-helper 2 (T. Using a mouse model of experimental allergic asthma, we investigated the efficacy of human adipose-derived mesenchymal stem cells (hADSCs) or human bone marrow-derived mesenchymal stem cells (hBMSCs) according to treatment frequency and timing.. Ovalbumin (OVA)-sensitized and -challenged mice exhibited airway hyperresponsiveness (AHR), airway inflammation, and significant increases in T. The results of treatment with hADSCs or hBMSCs suppresses AHR and airway inflammation. However, double hMSC treatment significantly induces eosinophilic airway inflammation and lung histological changes. Therefore, double hMSC treatment is ineffective against asthma and single injection frequency appears to be more important for the treatment of asthma. These results suggest that hMSC therapy can be used for treatment of asthma patients but that it should be used carefully.

Topics: Adipose Tissue; Animals; Asthma; Bone Marrow Cells; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Humans; Lung; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Severity of Illness Index

Accumulating evidence indicates that thrombin, the major effector of the coagulation cascade, plays an important role in the pathogenesis of asthma. Interestingly, dabigatran, a drug used in clinical anticoagulation, directly inhibits thrombin activity. The aim of this study was to investigate the effects and mechanisms of dabigatran on airway smooth muscle remodeling in vivo and in vitro. Here, we found that dabigatran attenuated inflammatory pathology, mucus production, and collagen deposition in the lungs of asthmatic mice. Additionally, dabigatran suppressed Yes-associated protein (YAP) activation in airway smooth muscle of asthmatic mice. In human airway smooth muscle cells (HASMCs), dabigatran not only alleviated thrombin-induced proliferation, migration and up-regulation of collagen I, α-SMA, CTGF and cyclin D1, but also inhibited thrombin-induced YAP activation, while YAP activation mediated thrombin-induced HASMCs remodeling. Mechanistically, thrombin promoted actin stress fibre polymerization through the PAR1/RhoA/ROCK/MLC2 axis to activate YAP and then interacted with SMAD2 in the nucleus to induce downstream target genes, ultimately aggravating HASMCs remodeling. Our study provides experimental evidence that dabigatran ameliorates airway smooth muscle remodeling in asthma by inhibiting YAP signalling, and dabigatran may have therapeutic potential for the treatment of asthma.

Topics: Actins; Adaptor Proteins, Signal Transducing; Airway Remodeling; Animals; Asthma; Biomarkers; Cell Cycle Proteins; Dabigatran; Disease Models, Animal; Fluorescent Antibody Technique; Immunohistochemistry; Lung; Male; Mice; Muscle, Smooth; Ovalbumin; Signal Transduction; Stress Fibers; Thrombin; YAP-Signaling Proteins

Asthma is a complex and heterogeneous inflammatory response characterized by various immune cells, including myeloid-derived suppressor cells (MDSCs) and CD4

Topics: Animals; Asthma; Cytokines; Gene Expression Regulation; Granulocytes; Lung; Male; Mice; Mice, Inbred BALB C; Myeloid-Derived Suppressor Cells; Ovalbumin; Random Allocation; Spleen; T-Lymphocytes, Regulatory; Th17 Cells

Microalgal exopolysaccharides (EPSs) are given great attention due to their potential biotechnology applications. Purified C. vulgaris EPS was subjected to compositional and sugar linkage analyses, and partial acid hydrolysis. Hydrolysate separation by gel chromatography afforded oligosaccharide fractions. Both, EPS and oligomers were studied by NMR spectroscopy. Data suggest very complex highly branched α-L-arabino-α-L-rhamno-α,β-D-galactan structure. Backbone repeating unit is formed by →2)-α-L-Rha (1 → 3)-α-L-Rha(1 → sequence, highly branched by long 1,6-linked α-D-Galp side chains, further branched at C2, C3 or C4 by α-L-Araf, α-D-Galf and β-D-Galf residues. α-L-Araf form longer 1,2-linked chains branched at C3, C4 or C5. Galf residues are localized as terminal units predominantly in the β configuration, while α-D-Galp and α-L-Araf may be partially O-methylated. Ex vivo biological assays showed increased interleukin-12 (IL-12) and interferon-gamma (INF-γ) levels corresponding to transforming growth factor beta (TGF-β) decrease in guinea pig model experimental asthma. These facts point to the anti-remodelling effect of Chlorella EPS and suggest its possible application in the treatment of asthma and chronic obstructive pulmonary disorder.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Budesonide; Chlorella vulgaris; Chromatography, Gel; Disease Models, Animal; Galactans; Guinea Pigs; Hydrolysis; Interferon-gamma; Interleukin-12; Magnetic Resonance Spectroscopy; Oligosaccharides; Ovalbumin; Transforming Growth Factor beta

As a chronic airway disease, asthma has two characteristics, tissue remodeling and airway inflammation. This research focused on miR-92a to explore how it works in asthma. We revealed that the expressions of miR-92a were decreased in both serum and lung tissues from ovalbumin-induced asthma mouse. Bioinformatics analysis, quantitative polymerase chain reaction (qPCR) and dual luciferase assay revealed that miR-92a targets MUC5AC, which was linked to mucus hypersecretion in the pulmonary tracts. By injecting miR-92a-mimics into the trachea, both the airway hyper-reactivity and airway inflammation can be alleviated in an asthma mouse model which is induced by ovalbumin. Moreover, the goblet cell phenotype of asthmatic mice is significantly reduced by the action of miR-92a. Furthermore, miR-92a blocked interleukin (IL)-13-induced MUC5AC luciferase activity in 16HBE. Together, upregulation of miR-92a expression in asthmatic mice plays a role in blocking goblet cell metaplasia by targeting MUC5AC, and thus in the treatment of chronic airway diseases, miR-92a can prevent epithelial remodeling, which is a reasonable method.

Topics: Animals; Asthma; Female; Gene Expression Regulation; Goblet Cells; Metaplasia; Mice; Mice, Inbred C57BL; MicroRNAs; Mucin 5AC; Ovalbumin

Topics: Animals; Apoptosis; Asthma; Biological Products; Biomarkers; Cytokines; Dendritic Cells; Female; Fungal Proteins; Immunohistochemistry; Lymphocyte Count; Mice; NF-kappa B; Ovalbumin; Pneumonia; Recombinant Proteins; Reishi; STAT3 Transcription Factor; T-Lymphocytes, Regulatory; Th17 Cells

The enhanced type 2 helper (Th2) immune response is responsible for the pathogenesis of allergic asthma. To suppress the enhanced Th2 immune response, activation of the Th1 immune response has been an alternative strategy for anti-asthma therapy. In this context, effective Th1-inducing adjuvants that inhibit the development of allergic asthma but do not flare the side effects of the primary agent are required in clinical treatment and preventive medicine.. In this study, we aimed to determine the regulation of the Th2 type immune response in asthma by a novel immunostimulatory oligodeoxynucleotide (ODN) derived from Cryptococcus neoformans, termed ODN112, which contains a cytosine-guanine (CG) sequence but not canonical CpG motifs.. Using an ovalbumin-induced asthma mouse model, we assessed the effect of ODN112 on prototypical asthma-related features in the lung and on the Th1/Th2 profile in the lymph nodes and lung of mice treated with ODN112 during sensitization.. ODN112 treatment attenuated asthma features in mice. In the bronchial lymph nodes of the lungs and in the spleen, ODN112 increased interferon-γ production and attenuated Th2 recall responses. In dendritic cells (DCs) after allergen sensitization, ODN112 enhanced cluster of differentiation (CD) 40 and CD80 expression but did not alter CD86 expression. Interleukin-12p40 production from DCs was also increased in a Th2-polarizing condition. Our results suggest that ODN112 is a potential Th1-inducing adjuvant during Th2 cell differentiation in the sensitization phase.

Topics: Allergens; Animals; Asthma; Cell Differentiation; CpG Islands; Cryptococcus neoformans; Dendritic Cells; Disease Models, Animal; Female; Humans; Hypersensitivity; Mice; Mice, Inbred C57BL; Oligodeoxyribonucleotides; Ovalbumin; Th1-Th2 Balance; Th2 Cells; Toll-Like Receptor 9

microRNA-146a has been reported to be a regulator in the process of attenuating asthma by inhibiting Toll-like receptor 2 (TLR2) pathway. This study aimed to investigate how miR146a-inhibitor affect the symptom of asthma and the underlying mechanisms.. Ovalbumin (OVA)-induced allergic asthma mice model was established by intraperitoneal injection with 20 μg of OVA. Total cells and differential inflammatory cells in bronchoalveolar lavage fluid were counted by flow cytometry. The expression levels of molecules and cytokines in TLR2 signaling pathway were detected by Q-PCR and ELISA.. miR146a-inhibitor attenuated OVA-induced allergic asthma by increasing Th1 cytokines in OVA-induced allergic asthma model, and the treatment of miR146a-inhibitor can reduce the inflammation caused by asthma, followed by the down-regulation of IL-5 and IL-13 in sorted ILC2. The inhibition of miR-146a significantly reduced symptoms of asthma model with TLR2-related molecules being up-regulated.. It was found that miR-146a is an important regulator in OVA-induced allergic asthma model, which can relieve symptoms of asthma through regulating TLR2 pathway. These findings provide a theoretical basis for solving asthma in clinical treatment.

Topics: Allergens; Animals; Asthma; Biomarkers; Combined Modality Therapy; Disease Models, Animal; Disease Susceptibility; Mice; MicroRNAs; Molecular Mimicry; Ovalbumin; RNA Interference; Signal Transduction; Th1 Cells; Toll-Like Receptor 2; Toll-Like Receptors

Asthma is a serious global health problem, severely affecting the lives of sufferers and their families. An exceptionally hygienic home and reduced microbial exposure can aggravate the incidence of childhood asthma.. Specific-pathogen-free BALB/c mice were pre-treated with bacterial lysate (BL; 1 mg/kg) as a high microbial load maternal mouse model, and then, the offspring mice were established as an allergic airway disease (AAD) model. The expression levels of TLR2, TLR4, and HDAC9 in the mother's intestine and the offspring's lungs were detected. Relevant indicators of regulatory T cells (Tregs) were identified in the mother and offspring mice. The changes in the expression of Th1-, Th2-, Th9-, and Th17-related cytokines in the offspring mice were evaluated among different pre-treated groups.. After augmenting the mothers' intestinal microbiota through oral BL gavage, the expression of TLR2 and TLR4 in the colon mucosa and colon lymphoid tissues was enhanced and that of HDAC9 in the colon mucosa was decreased, and the proportion of spleen Tregs was increased. The offspring showed similar changes in the AAD model compared with the offspring of the control-group mothers: TLR2 and TLR4 expression in the lungs and the proportion of spleen Tregs increased, HDAC9 expression in the lungs decreased, and AAD-induced airway pathologic characteristics were reversed; additionally, Th1/Th2 and Th9 imbalances were rectified.. This study presents a new framework for the prevention of childhood asthma, elucidating the mechanism of regulating the mother's intestinal microbiome to protect the offspring's early asthma via animal experiments.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Humans; Hypersensitivity; Lung; Mice; Mice, Inbred BALB C; Microbiota; Ovalbumin; Th2 Cells

MicroRNA (miRNA) mimics or antagomirs hold great promise for asthma treatment compared with glucocorticoids as mainstay therapy for asthma. But the role of miRNA in regulating asthmatic inflammation is largely unclear. We previously reported that miR-3162-3p in the peripheral blood of children with asthma was obviously upregulated compared to that in healthy children. This study aimed to elucidate the role of miR-3162-3p in pulmonary inflammation in normal and asthmatic mice as well as preliminarily explore the potential of miR-3162-3p antagomir in asthma treatment. A noninvasive whole-body plethysmograph measured airway responsiveness. Both qRT-PCR and Western blot were used to detect the expression of miRNA, mRNA, or protein. Cells in bronchoalveolar lavage fluid were counted by platelet counting and Wright's staining. Inflammatory infiltration and mucus secretion were identified by hematoxylin and eosin and periodic acid-Schiff  staining, respectively. Cytokines in the lungs were detected by ELISA. The miR-3162-3p mimic intraperitoneally administered to normal mice decreased β-catenin levels in the lungs without obviously altering the lung histology and cytokine levels. Antagonizing miR-3162-3p in ovalbumin-induced asthmatic mice effectively alleviated the typical features of asthma, such as airway hyper-responsiveness, airway inflammation, and Th1/Th2 cytokine imbalance, and concomitantly rescued the total and active β-catenin expression. Collectively, we discovered divergent roles of miR-3162-3p in lung inflammation between normal and asthmatic mice. The anti-inflammatory effects of the miR-3162-3p antagomir were comparable to those of glucocorticoid treatment. Our study helped in understanding the contribution of miRNAs to the pathogenesis of asthma.

Topics: Allergens; Animals; Antagomirs; Asthma; beta Catenin; Child; Cytokines; Disease Models, Animal; Gene Expression Regulation; Humans; Lung; Mice; MicroRNAs; Ovalbumin; Pneumonia; Respiratory Hypersensitivity; Th1 Cells; Th2 Cells

Titanium dioxide nanoparticles (TiO. According to the results, the mice receiving OVA alone or OVA plus TiO. Based on the findings of the current study, intranasally or inhalation exposure to high-dose nanosized TiO

Topics: Administration, Intranasal; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cytokines; Eosinophils; Female; Inhalation Exposure; Mice; Mice, Inbred BALB C; Nanoparticles; Ovalbumin; Respiratory Function Tests; T-Lymphocytes, Helper-Inducer; Titanium

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Mice; Ovalbumin; Oxidative Stress; Plant Exudates; Plant Stems; Sambucus nigra; Spleen; Th2 Cells

The cholinergic anti-inflammatory pathway has been shown to regulate lung inflammation and cytokine release in acute models of inflammation, mainly via α7 nicotinic receptor (α7nAChR). We aimed to evaluate the role of endogenous acetylcholine in chronic allergic airway inflammation in mice and the effects of therapeutic nAChR stimulation in this model. We first evaluated lung inflammation and remodeling on knock-down mice with 65% of vesicular acetylcholine transport (VAChT) gene reduction (KDVAChT) and wild-type(WT) controls that were subcutaneously sensitized and then inhaled with ovalbumin(OVA). We then evaluated the effects of PNU-282987(0.5-to-2mg/kg),(α7nAChR agonist) treatment in BALB/c male mice intraperitoneal sensitized and then inhaled with OVA. Another OVA-sensitized-group was treated with PNU-282987 plus Methyllycaconitine (MLA,1 mg/kg, α7nAChR antagonist) to confirm that the effects observed by PNU were due to α7nAChR. We showed that KDVAChT-OVA mice exhibit exacerbated airway inflammation when compared to WT-OVA mice. In BALB/c, PNU-282987 treatment reduced the number of eosinophils in the blood, BAL fluid, and around airways, and also decreased pulmonary levels of IL-4,IL-13,IL-17, and IgE in the serum of OVA-exposed mice. MLA pre-treatment abolished all the effects of PNU-282987. Additionally, we showed that PNU-282987 inhibited STAT3-phosphorylation and reduced SOCS3 expression in the lung. These data indicate that endogenous cholinergic tone is important to control allergic airway inflammation in a murine model. Moreover, α7nAChR is involved in the control of eosinophilic inflammation and airway remodeling, possibly via inhibition of STAT3/SOCS3 pathways. Together these data suggest that cholinergic anti-inflammatory system mainly α7nAChR should be further considered as a therapeutic target in asthma.

Topics: Airway Remodeling; Allergens; alpha7 Nicotinic Acetylcholine Receptor; Animals; Asthma; Benzamides; Bridged Bicyclo Compounds; Bronchoalveolar Lavage Fluid; Chronic Disease; Cytokines; Disease Models, Animal; Inflammation; Leukocyte Count; Lung; Male; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; STAT3 Transcription Factor; Suppressor of Cytokine Signaling 3 Protein; Vesicular Acetylcholine Transport Proteins

To observe the efficacy of San'ao Decoction(SAD) in diffusing the lung and relieving asthma, and its intervention effect on the expression of transient receptor potential V2(TRPV2) during alleviating asthma, this study replicated an ovalbumin(OVA)-induced asthmatic mice model, and investigated the intervention effect of SAD on the airway inflammation and airway hyperresponsiveness. The regulatory mechanisms of SAD on the mRNA and protein expressions of TRPV2 in lung tissues and the levels of interleukin-4(IL-4),-10(IL-10), nerve growth factor(NGF), prostaglandin D_2(PGD_2) in bronchoalveolar lavage fluid(BALF) were discussed. Compared with the control group, the model group showed typical asthmatic phenotype, the level of eosinophils(EOS) in peripheral blood and BALF as well as the airway hyperresponsiveness were increased(P<0.01), and pathological damage in lung tissue was serious. The mRNA and protein expressions of TRPV2 in lung tissue were increased significantly, while the levels of IL-4, IL-10, NGF and PGD_2 in BALF were elevated(P<0.05,P<0.01). SAD could relieve bronchial asthma manifested as repaired lung patholo-gical changes(P<0.05), reduce the level of EOS in blood and BALF(P<0.05, P<0.01), and improve pulmonary resistance and lung compliance(P<0.05, P<0.01). SAD could also regulate the inflammatory cytokine levels of IL-4, IL-10, NGF, PGD_2 in BALF, and reduce the gene and protein expression of TRPV2 in the lung tissue(P<0.05, P<0.01). It is verified that SAD could reduce the lung inflammation, and improve lung function in asthmatic mice. The regulatory mechanism of SAD on asthma induced by OVA might be related to the regulation of TRPV2 expression and the induced decrease of Th2-related cytokines and neuropeptides, which provides the evidences for the treatment of asthma with SAD.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Calcium Channels; Disease Models, Animal; Drugs, Chinese Herbal; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; TRPV Cation Channels

The interleukin 17 receptor E (IL-17RE) is specific for the epithelial cytokine interleukin-17C (IL-17C). Asthma exacerbations are frequently caused by viral infections. Polyinosinic:polycytidylic acid (pIC) mimics viral infections through binding to pattern recognition receptors (e.g. TLR-3). We and others have shown that pIC induces the expression of IL-17C in airway epithelial cells. Using different mouse models, we aimed to investigate the function of IL-17RE in the development of experimental allergic asthma and acute exacerbation thereof.. Wild-type (WT) and IL-17RE deficient (Il-17re. Ablation of IL-17RE did not affect the development of OVA-induced allergic airway inflammation and AHR. pIC induced inflammation independent of IL-17RE in the absence of allergic airway inflammation. Treatment of mice with pIC exacerbated pulmonary inflammation in sensitized and OVA-challenged mice in an IL-17RE-dependent manner. The pIC-induced expression of cytokines (e.g. keratinocyte-derived chemokine (KC), granulocyte-colony stimulating factor (G-CSF)) and recruitment of neutrophils were decreased in Il-17re. Our results indicate that IL-17RE mediates virus-triggered exacerbations but does not have a function in the development of allergic lung disease.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Epithelial Cells; Inflammation Mediators; Interleukin-17; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophil Infiltration; Ovalbumin; Poly I-C; Receptors, Interleukin-17; Respiratory Hypersensitivity; Respiratory Mucosa

To clarify the effects of dietary supplementation of protocatechuic acid (PCA) and in-depth mechanisms on allergic asthma in ovalbumin (OVA)-induced mice.. Female BALB/c mice were randomly divided into three groups (n = 10 in each group): control group, OVA-induced allergic asthma group, and OVA plus PCA group.. Dietary supplementation of PCA was achieved by adding 50 mg/kg PCA to AIN 93G diet for 25 days.. Peripheral blood cells, pulmonary inflammatory cell infiltration, the levels of IL-4, IL-5, and IL-13 in bronchoalveolar lavage fluid (BALF), the mRNA levels of Th2-related genes in the lungs, and the protein expressions of the IL-4Rα-STAT6 and the Jagged1/Jagged2-Notch1/Notch2 signaling pathways were measured.. Significantly reduced inflammatory cells infiltration and mucosal hypersecretion in the lung tissues, repaired levels of interleukin IL-4, IL-5, and IL-13 in the BALF, and decreased mRNA expression of IL-4, IL-5, and GATA3 were observed in OVA plus PCA group. Moreover, PCA treatment down-regulated the protein levels of IL-4Rα-STAT6 and Jagged1/Jagged2-Notch1/Notch2 signaling pathways.. Dietary supplement of PCA alleviated allergic asthma partly through suppressing the IL-4Rα-STAT6 and Jagged1/Jagged2-Notch1/Notch2 signaling pathways in mice. Our study provided the theoretic basis of PCA used as functional food in preventing allergic asthma.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dietary Supplements; Female; Functional Food; Hydroxybenzoates; Jagged-1 Protein; Jagged-2 Protein; Leukocyte Count; Lung; Mice, Inbred BALB C; Ovalbumin; Receptor, Notch1; Receptor, Notch2; Receptors, Cell Surface; Signal Transduction; STAT6 Transcription Factor

The ovalbumin-induced (OVA) chronic allergic airways murine model is a well-established model for investigating pre-clinical therapies for chronic allergic airways diseases, such as asthma. Here, we examined the effects of several experimental compounds with potential anti-asthmatic effects including resveratrol (RV), relaxin (RLN), L-sulforaphane (LSF), valproic acid (VPA), and trichostatin A (TSA) using both a prevention and reversal model of chronic allergic airways disease. We undertook a novel analytical approach using focal plane array (FPA) and synchrotron Fourier-transform infrared (S-FTIR) microspectroscopic techniques to provide new insights into the mechanisms of action of these experimental compounds. Apart from the typical biological effects, S-FTIR microspectroscopy was able to detect changes in nucleic acids and protein acetylation. Further, we validated the reduction in collagen deposition induced by each experimental compound evaluated. Although this has previously been observed with conventional histological methods, the S-FTIR technique has the advantage of allowing identification of the type of collagen present. More generally, our findings highlight the potential utility of S-FTIR and FPA-FTIR imaging techniques in enabling a better mechanistic understanding of novel asthma therapeutics.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Chronic Disease; Disease Models, Animal; Drug Evaluation, Preclinical; Female; Hydroxamic Acids; Isothiocyanates; Mice; Mice, Inbred BALB C; Ovalbumin; Relaxin; Resveratrol; Spectroscopy, Fourier Transform Infrared; Sulfoxides; Synchrotrons; Treatment Outcome; Valproic Acid

The purpose of this study was to explore the potential contracting effect of leptin on isolated guinea pig tracheal smooth muscle (TSM), the possible mechanism, and the impact of epithelium denudation or allergen sensitization, respectively. An in vitro experiment investigated the effect of leptin at a concentration of 250-1000 nmol/L on isolated guinea pig TSM with an intact or denuded epithelium. Ovalbumin and IgE were used to test the impact of active and passive sensitization. The isolated TSM strips were incubated in Krebs solution and aerated with carbogen (95% O

Topics: Animals; Asthma; Disease Models, Animal; Guinea Pigs; Humans; Leptin; Male; Muscle Contraction; Muscle, Smooth; Ovalbumin; Receptors, Leptin; Trachea

Asthma is a common chronic inflammatory airway disease. Recent studies have reported that interleukin (IL)‑33 is a potential link between the airway epithelium and Th2‑type inflammatory responses, which are closely related to the progression of asthma. The IL‑33 receptor, ST2, is highly expressed in group 2 innate lymphoid cells (ILC2s), Th2 cells, mast cells, eosinophils and natural killer (NK) cells. Cnidii Fructus is a Chinese herb with a long history of use in the treatment of asthma in China. Osthole is one of the major components of Cnidii Fructus. The present study examined the anti‑asthmatic effects of osthole in mice and aimed to elucidate the underlying mechanisms involving the IL‑33/ST2 pathway. BALB/c mice were sensitized and challenged with ovalbumin and then treated with an intraperitoneal injection of osthole (25 and 50 mg/kg). Subsequently, the airway hyper‑responsiveness (AHR) and inflammation of the lungs were evaluated. The amounts of IL‑4, IL‑5, IL‑13, interferon (IFN)‑γ and IL‑33 in the bronchoalveolar lavage fluid (BALF) were measured by Luminex assay and their mRNA levels in the lungs were measured by reverse transcription‑quantitative PCR. The histopathology of the lungs was performed with H&E, PAS and Masson's staining. The expression of ST2 in the lungs was evaluated by immunohistochemistry. The data demonstrated that osthole markedly reduced AHR and decreased the number of eosinophils and lymphocytes in BALF. It was also observed that osthole significantly inhibited the release of Th2‑type cytokines (IL‑4, IL‑5 and IL‑13) and upregulated the IFN‑γ level in BALF. Moreover, osthole significantly attenuated the IL‑33 and ST2 expression in the lungs of asthmatic mice. On the whole, osthole attenuated ovalbumin‑induced lung inflammation through the inhibition of IL‑33/ST2 signaling in an asthmatic mouse model. These results suggest that osthole is a promising target for the development of an asthma medication.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Coumarins; Disease Models, Animal; Drugs, Chinese Herbal; Female; Gene Expression Regulation; Inflammation; Interferon-gamma; Interleukin-1 Receptor-Like 1 Protein; Interleukin-33; Interleukins; Lung; Lymphocyte Count; Mice; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Pulmonary Eosinophilia; Random Allocation; RNA, Messenger; Signal Transduction

Sirtuin 1 (SIRT1) is involved in the pathogenesis of allergic asthma. This study aimed to investigate whether EX‑527, a specific SIRT1 inhibitor, exerted suppressive effects on allergic airway inflammation in mice submitted to ovalbumin (OVA) inhalation. In addition, this study assessed whether such a protective role was mediated by autophagy suppression though mammalian target of rapamycin (mTOR) activation. Female C57BL/6 mice were sensitized to OVA and EX‑527 (10 mg/kg) was administered prior to OVA challenge. The study found that EX‑527 reversed OVA‑induced airway inflammation, and reduced OVA‑induced increases in inflammatory cytokine expression, and total cell and eosinophil counts in bronchoalveolar lavage fluid. In addition, EX‑527 enhanced mTOR activation, thereby suppressing autophagy in allergic mice. To assess whether EX‑527 inhibited airway inflammation in asthma through the mTOR‑mediated autophagy pathway, rapamycin was administered to mice treated with EX‑527 after OVA sensitization. All effects induced by EX‑527, including increased phosphorylated‑mTOR and decreased autophagy, were abrogated by rapamycin treatment. Taken together, the present findings indicated that EX‑527 may inhibit allergic airway inflammation by suppressing autophagy, an effect mediated by mTOR activation in allergic mice.

Topics: Administration, Inhalation; Animals; Asthma; Autophagy; Carbazoles; Cytokines; Disease Models, Animal; Down-Regulation; Female; Mice; Mice, Inbred C57BL; Ovalbumin; Phosphorylation; Sirolimus; Sirtuin 1; TOR Serine-Threonine Kinases

The aim of the present study was to establish an integrated network of DNA methylation and RNA expression in an ovalbumin (OVA)‑induced asthma model, and to investigate the epigenetically‑regulated genes involved in asthma development. Genome‑wide CpG‑DNA methylation profiling was conducted through the use of a methylated DNA immunoprecipitation microarray and RNA sequencing was performed using three lung samples from mice with OVA‑induced asthma. A total of 35,401 differentially methylated regions (DMRs) were identified between mice with OVA‑induced asthma and control mice. Of these, 3,060 were located in promoter regions and 370 of the genes containing these DMRs demonstrated an inverse correlation between methylation and gene expression. Kyoto Encyclopedia of Genes and Genomes pathway analysis identified that 368 genes were upregulated or downregulated in OVA‑induced asthma samples, including genes involved in 'chemokine signalling pathway', 'focal adhesion', 'leukocyte transendothelial migration' and 'vascular smooth muscle contraction signaling' pathways. Integrated network analysis identified four hub genes, consisting of three upregulated genes [forkhead box O1 (FOXO1), SP1 transcription factor (SP1) and amyloid β precursor protein (APP)], and one downregulated gene [RUNX family transcription factor 1 (RUNX1)], all of which demonstrated an association between DNA methylation and gene expression. These genes were highly interconnected nodes in the Ingenuity Pathway Analysis module and were functionally significant. A total of four interconnected hub genes, FOXO1, RUNX1, SP1 and APP, were identified from the integrated DNA methylation and gene expression networks involved in asthma development. These results suggested that modulating these four genes could effectively control the development of asthma.

Topics: Animals; Asthma; Disease Models, Animal; DNA Methylation; Down-Regulation; Epigenesis, Genetic; Female; Gene Expression Profiling; Gene Expression Regulation; Gene Regulatory Networks; Genome-Wide Association Study; Humans; Mice; Ovalbumin; Sequence Analysis, DNA; Up-Regulation

Epidemiological studies and mice models have demonstrated that air pollution containing particulate matter smaller than 2.5 μm (PM2.5) exacerbates acute episodes of asthma in both children and adults.. To investigate the effect of continuous PM2.5 treatment on asthma regulation mechanism behind this effect.. In this study, the effects of continuous exposure to PM2.5 on asthma and eosinophil recruitment was compared to the effect of a single pre-ovalbumin (OVA)-sensitization exposure to PM2.5. Wild-type mice were either challenged once with PM2.5 + OVA before sensitization and asthma induction over a 27-day period, or with 5 times of PM2.5 + OVA treatment and sensitization/asthma induction over the same period.. Continuous exposure to PM2.5 significantly increased total plasma immunoglobulin E (IgE), bronchial alveolar lavage fluid (BALF) cell numbers, eosinophils, and macrophages, leading to increased lung injury. This effect was regulated through increased production of chemokines and cytokines, such as interleukin (IL)-1β, monocyte chemoattractant protein 1 (MCP-1), IL-12, IL-5, IL-13, and prostaglandin D2 (PGD2). Eosinophil recruitment during continuous PM2.5 treatment was regulated through phosphorylation of the JAK/STAT6 pathway. As this study shows, continuous PM2.5 treatment significantly worsens asthma as compared to single exposure to PM2.5 or OVA exposure alone.. Our findings reveal that continuous exposure of PM2.5 exacerbates OVA-induced asthma in mouse lung through JAK-STAT6 signaling pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Janus Kinases; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Signal Transduction; STAT6 Transcription Factor

Epidemiological and experimental studies suggest that microbial exposure in early childhood is linked with reduced risk to suffer asthma. Thus microbial components with immunoregulatory capabilities might serve as a preventive strategy for allergic asthma. Recently, it was identified that Streptococcus pneumoniae aminopeptidase N (PepN) could suppress T cell effector function. We sought to investigate the effect of PepN on murine allergic asthma and elucidate the underlying mechanism.. The effects of intranasal administration of PepN during or before sensitization were examined in ovalbumin (OVA)-induced murine allergic asthma. The roles of CD11b. Administration of PepN during or before sensitization attenuated type-2 airway inflammation (eosinophilia, mucus hypersecretion, Th2 cytokines production and IgE production) in allergic asthma mice. PepN reduced lung accumulation of CD11b. PepN alleviated type-2 inflammation in OVA-induced allergic asthma mice, which was mediated by regulation of lung CD11b

Topics: Animals; Asthma; CD13 Antigens; Child, Preschool; Cytokines; Dendritic Cells; Disease Models, Animal; Humans; Immunity, Innate; Inflammation; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Ovalbumin; Streptococcus pneumoniae

Sojadodamgangki-tang (SDG) is a traditional East-Asian herbal medicine mainly composed of Pinellia ternate (Thunb.) Makino, Perilla frutescens (L.) Britt and 10 kinds of medicinal herbs. It has been used to treat asthma and mucus secretion including lung and bronchi.. The aim of this study was to investigate the anti-inflammatory effects of Sojadodamgangki-tang (SDG) on allergic lung inflammation in vitro and in vivo as well as the underlying mechanisms.. We used an ovalbumin (OVA)-induced murine allergic airway inflammation model. Five groups of 8-week-old female BALB/C mice were divided into the following groups: saline control group, the vehicle (allergic) group that received OVA only, groups that received OVA and SDG (200 mg/kg or 400 mg/kg), and a positive control group that received OVA and Dexamethasone (5 mg/kg). In vitro experiments include T helper 2 (TH2) polarization system, murine macrophage cell culture, and human bronchial epithelial cell line (BEAS-2B) culture.. SDG administration reduced allergic airway inflammatory cell infiltration, especially of eosinophils, mucus production, Th2 cell activation, OVA-specific immunoglobulin E (IgE), and total IgE production. Moreover, the activation of alveolar macrophages, which leads to immune tolerance in the steady state, was promoted by SDG treatment. Interestingly, SDG treatment also reduced the production of alarmin cytokines by the human bronchial epithelial cell line BEAS-2B stimulated with urban particulate matter.. Our findings indicate that SDG has potential as a therapeutic drug to inhibit Th2 cell activation and promote alveolar macrophage activation.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Female; Macrophages, Alveolar; Mice; Mice, Inbred BALB C; Ovalbumin; Perilla; Pinellia; Th2 Cells

Panax notoginseng saponin R1 (PNS-R1) is one of the most important chemical monomers derived from the panax notoginseng, and our previous study found that PNS-R1 reduced glucocorticoid-induced apoptosis in asthmatic airway epithelial cells. Thus, in this study, we explored the effects of the PNS-R1 on inflammation of allergic asthma.. The asthmatic mice were administered 15 mg/kg PNS-R1 by intraperitoneal injection three days before sensitized to OVA. The effects of PNS-R1 on asthmatic mice were detected by airway hyperresponsiveness, inflammation, mucus hypersecretion and inflammatory cytokines such as interleukin (IL)-13, IL-4, IL-5, IL-8 and tumor necrosis factor (TNF)-α were studied. We also treated human bronchial epithelial cells (16HBE) with house dust mites (HDM) and then detected the secretion of cellular inflammatory factors (IL-13 and TNF-α). Western blot and immunofluorescence were used to examine the effect of PNS-R1 on TNF-α/NF-κB pathway. TNF-α/NF-κB/IKK signal pathway activator was used in PNS-R1-treated asthmatic mice.. PNS-R1 significantly reduced the airway inflammatory, mucus secretion and hyperresponsiveness in asthma model. It also reduced the levels of IL-13, IL-4, IL-5 and IL-8 in bronchoalveolar lavage fluid (BALF) and IgE and OVA-specific IgE in serum for asthma mice. PNS-R1 reduced IL-13 and TNF-α secretion in HDM-treated 16HBE cells. In addition, PNS-R1 suppressed TNF-α/NF-κB pathway in both asthmatic mice and 16HBE. Activation of NF-kB pathway reversed the therapeutic effect of PNS-R1 on asthmatic mice.. The results indicated that PNS-R1 effectively suppresses allergic airway inflammation of asthma partly through TNF-α/NF-κB pathway. PNS-R1 may play a potential role in allergic asthma treatment in the future.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cell Line; Disease Models, Animal; Female; Humans; I-kappa B Kinase; Immunoglobulin E; Inflammation; Lung; Male; Mice, Inbred C57BL; Mucin 5AC; Mucus; NF-kappa B; Ovalbumin; Panax notoginseng; Pyroglyphidae; Respiratory Hypersensitivity; Saponins; Signal Transduction; Tumor Necrosis Factor-alpha

Numerous population studies conducted worldwide indicate that the prevalence of asthma is higher in obese versus lean individuals. It has been reported that sensitized lean mice has a better recovery of lung inflammation in asthma. Extracellular matrix (ECM) plays an essential role in the structural support of the lungs regulating the airways diameter, thus preventing its collapse during expiration. ECM renewal by metalloproteinase (MMPs) enzymes is critical for pulmonary biology. There seems to be an imbalance of MMPs activity in asthma and obesity, which can impair the lung remodeling process. In this study, we characterized the pulmonary ECM of obese and lean mice, non-sensitized and sensitized with ovalbumin (OVA). Pharmacological intervention was performed by using anti-TNF-α, and MMP-8 and MMP-9 inhibitors in obese and lean sensitized mice. Activity of MMPs was assessed by gelatinase electrophorese, western blotting and zymogram in situ. Unbalance of MMP-2, MMP-8, MMP-9 and MMP-12 was detected in lung tissue of OVA-sensitized obese mice, which was accompanied by high degradation, corroborating an excessive deposition of types I and III collagen in pulmonary matrix of obese animals. Inhibitions of TNF-α and MMP-9 reduced this MMP imbalance, clearly suggesting a positive effect on pulmonary ECM. Obese and lean mice presented diverse phenotype of asthma regarding the ECM compounds and the inhibition of MMPs pathway could be a good alternative to regulate the activity in ECM lungs of asthmatic obese individuals.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Extracellular Matrix; Inflammation; Lung; Male; Matrix Metalloproteinase 8; Matrix Metalloproteinase 9; Metalloproteases; Mice; Mice, Inbred C57BL; Obesity; Ovalbumin; Protease Inhibitors; Tumor Necrosis Factor-alpha

Increased number of airway smooth muscle cells (ASMCs) is a characteristic of airway remodeling in asthma. In this study we investigated whether emodin alleviated airway remodeling in a murine asthma model and reduced the proliferation of ASMCs in vitro. We provided in vivo evidence suggesting that intraperitoneal injection of emodin (20 mg/kg) 1 h prior to OVA challenge apparently alleviated the thickness of airway smooth muscle, the mass of alpha-smooth muscle actin (α-SMA), collagen deposition, epithelial damage, goblet cell hyperplasia, airway inflammation and airway hyperresponsiveness (AHR) in lung tissue. Meanwhile, we found that emodin suppressed the activation of the Akt pathway in lungtissue of allergic mouse models. Additionally, we found that emodin inhibited cellular proliferation and Akt activation in a dose-dependent manner in vitro. Furthermore, LY294002, an inhibitor for PI3K, abrogated serum-induced phosphorylation of Akt, and decreased the proliferation of ASMCs. These findings indicated that emodin alleviated ASMCs proliferation by inhibiting PI3K/Akt pathway in vivo and in vitro, which may provide a potential therapeutic option for airway smooth muscle remodeling in asthma.

Topics: Actins; Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Proliferation; Cells, Cultured; Collagen; Cytokines; Disease Models, Animal; Emodin; Eosinophils; Female; Goblet Cells; Leukocytes; Lung; Mice, Inbred BALB C; Myocytes, Smooth Muscle; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Respiratory Hypersensitivity; Th2 Cells

Asthma is a consequence of complex gene-environment interactions. Exploring the heterogeneity of asthma in different stages is contributing to our understanding of its pathogenesis and the development of new therapeutic strategies, especially in severe cases.. This study aimed to further understand the relationship between manifestations of acute and chronic asthma and various endotypes, and explore the severity of lung inflammation, cell types, cytokine/chemokine differences, and the effects of FIP-fve.. Acute and chronic OVA-sensitization mouse asthma models, based on our previously published method, were used and FIP-fve was used to evaluate the effect on these two models. BALF cytokines/chemokines were detected according to the manufacturer's protocol.. Seventeen cytokine/chemokine secretions were higher in the chronic stage than in the acute stage. Whether in acute stage or chronic stage, the FIP-fve treatment groups had reduced airway hyperresponsiveness, infiltration of airway inflammatory cells, secretion of cytokines, chemokines by Th2 cells, and TNF-α, IL-8, IL-17, CXCL-1, CXCL-10, CCL-17, and CCL-22, and it was also found that the Treg cell cytokine IL-10 had increased significantly. PCA (Principal Component Analysis) was also used to compare statistics and laboratory data to find the important biomarkers in different stages and after treatment with FIP-fve.. There are many different immune responses in the different stages of the asthma process. Drug treatment at the appropriate times might help reduce the worsening of asthma.

Topics: Airway Resistance; Animals; Asthma; Biomarkers; Cytokines; Desensitization, Immunologic; Disease Models, Animal; Female; Fungal Proteins; Immunoglobulin E; Immunoglobulin G; Mice; Mice, Inbred BALB C; Ovalbumin; Principal Component Analysis

Asthma is an inflammatory disease of the airways of the lungs, which is characterized by airflow obstruction and bronchospasms. Glabridin is a major flavonoid, especially found in root of Glycyrrhiza glabra, and has several pharmacological activities, including antioxidant and anti-inflammatory effects. The anti-asthmatic effect and possible mechanism of glabridin, however, have not been revealed so far. The aim of this study is to investigate the effects and possible mechanisms of glabridin against ovalbumin (OVA)-induced airway hyperresponsiveness (AHR) and inflammation in mice. In male BALB/c mice, asthma was induced by intraperitoneal (i.p) injection of OVA mixed with 2 mg aluminium hydroxide on days 0, 14 and boosted with OVA aerosol challenge on days 21, 22, and 23. Mice were either treated with dexamethasone (i.p, 1 mg/kg) or glabridin (10, 20, and 30 mg/kg) from days 18-23. Pulmonary function parameters such as peak inspiratory flow, peak expiratory flow, tidal volume, expiratory volume, the frequency of breathing, enhanced pause values were evaluated by using whole-body plethysmography. Measurements were performed at baseline and following methacholine (50 mg/mL) challenges. In addition, white blood cells (WBC) count, total protein, and IgE levels were measured in bronchial alveolar lavage fluid (BALF), lung, and serum, respectively. Glabridin (20 or 30 mg/kg) significantly attenuated (p < 0.05) OVA-induced alteration in respiratory parameters. Elevated counts of total WBC, differential WBC (neutrophils, lymphocytes, monocytes, and eosinophils) in BALF and the total protein in lungs and BALF were significantly decreased (p < 0.05) by glabridin (20 or 30 mg/kg). It also significantly attenuated the increased serum IgE levels (p < 0.05). As glabridin reduces the level of serum IgE, the total protein and the count of WBC and improves respiratory function, it may be a novel therapeutic agent in asthma.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Isoflavones; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Phenols

Asthma is an airway chronic inflammatory disease with significant morbidity, mortality and huge social economic burden. Previous research demonstrated that the root of Aster tataricus (RA) may have the potential to treat asthma, but the efficacy and mechanism were not clear. In this study, preliminary results in vitro showed that Fr-75 eluted from RA extract could not only completely inhibit the tracheal ring contraction raised by KCl in 20 min, but also effectively affect the tracheal ring contraction induced by KCl-, Ach- and His in a concentration-dependent manner (3.91-250 μg/mL). Further results on cells exhibited that Fr-75 could decrease the concentration of intracellular Ca

Topics: Animals; Anti-Asthmatic Agents; Aster Plant; Asthma; Bronchoalveolar Lavage Fluid; Calcium Channel Blockers; Guinea Pigs; Histamine Antagonists; In Vitro Techniques; Lung; Mice; Muscarinic Antagonists; Muscle Contraction; Myocytes, Smooth Muscle; Ovalbumin; Plant Extracts; Trachea; Tracheitis

Tetrahydrocurcumin (THC) is the major active metabolite of curcumin, which is a dietary factor derived from Curcuma species. Our previous study demonstrated a significant beneficial effect of THC in mice with allergic asthma. Glucocorticosteroids (GCs) are commonly used drugs in asthma. Whether THC supplementation could promote the beneficial effects of GC therapy on asthma has not yet been reported. The current study aimed to investigate the combined efficacy of GC and THC treatment in a mouse model of allergic asthma.. BALB/c mice were randomly divided into 5 groups: the control group, ovalbumin (OVA)-induced group, and OVA-induced mice treated with dietary THC only, intraperitoneal injection of dexamethasone (DEX) only, or THC combined with DEX. The nasal symptoms, histopathological alterations of lung tissues, lung cytokine production, and Th cell subsets were assessed.. THC or DEX had beneficial effects on nasal symptoms and pathological lung changes, and the therapeutic effects between THC and DEX treatment were comparable. Importantly, compared to the monotherapy groups (THC or DEX only), the combination of THC and DEX showed a significantly reduced nasal rubbing frequency, lower mucus hyperproduction, lower Th2 and Th17 cell numbers as well as lower related cytokine levels (IL-4, IL-5, and IL-17A).. Supplementation with THC can enhance the therapeutic effects of DEX to alleviate airway symptoms, lung inflammation, and the Th2 response. Our findings suggest that dietary administration of THC could act as an add-on therapy for asthma treated with GCs.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Asthma; Curcuma; Curcumin; Dexamethasone; Dietary Supplements; Disease Models, Animal; Female; Humans; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Animal models of asthma have shown that limonene, a naturally occurring terpene in citrus fruits, can reduce inflammation and airway reactivity. However, the mechanism of these effects is unknown. We first performed computational and molecular docking analyses that showed limonene could bind to both A

Topics: Animals; Asthma; Disease Models, Animal; Inflammation; Limonene; Lung; Mice; Mice, Transgenic; Ovalbumin; Receptor, Adenosine A2A

Bronchoconstriction was recently shown to cause airway remodeling and induce allergic airway inflammation in asthma. However, the mechanisms how mechanical stress via bronchoconstriction could induce airway inflammation and remodeling remain unclear.. We investigated the effect of bronchoconstriction induced by methacholine inhalation in a murine model of asthma.. BALB/c female mice were sensitized and challenged with ovalbumin (OVA), followed by treatment with methacholine by a nebulizer twice a day for 7 days. Twenty-four hours after the last methacholine treatment, the bronchoalveolar lavage fluid (BALF) and lung tissues were collected. The BALF was analyzed for total and differential cell counts and cytokine levels. The lung tissues were analyzed for goblet cell metaplasia, thickness of the smooth muscle, and lung fibrosis. The expression of cytokines in the lung was also examined.. OVA sensitization and challenge induced infiltration of total cells, macrophages, and eosinophils in the BALF along with goblet cell metaplasia and increased airway smooth muscle hypertrophy. Seven days after the last OVA challenge, untreated mice achieved reduction in airway inflammation, while methacholine maintained the number of BALF total cells, macrophages, and eosinophils. The percentage of goblet cells and the thickness of airway smooth muscle were also maintained by methacholine. Moreover, the treatment of methacholine induced the expression of transforming growth factor (TGF)-β in the lung. This result indicates that the production of TGF-β is involved in induction of airway remodeling caused by bronchoconstriction with methacholine.. Repeated bronchoconstriction caused by methacholine inhalation elicited allergic airway inflammation and airway remodeling.

Topics: Administration, Inhalation; Allergens; Animals; Asthma; Bronchoconstriction; Disease Models, Animal; Eosinophils; Female; Humans; Lung; Macrophages; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Transforming Growth Factor beta

Asthma is a chronic respiratory disease orchestrated by immune and structural cells. Identification of novel therapeutic strategies are needed for asthma due to the limitations of existing therapies. We have validated the anti-inflammatory, anti-asthmatic and immunomodulatory therapeutic properties of herbal decoction, Divya-Swasari-Kwath (DSK) using mouse model of ovalbumin (OVA) induced allergic asthma.. HPLC analysis identified the presence of Rutin, Glycyrrchzin, Gallic acid, Cinnamic acid, Chlorogenic acid, Caffeic acid and Piperine as bioactive herbal metabolites in DSK. Therapeutic treatment with herbal decoction DSK significantly alleviated the pathological features of allergic asthma including inflammatory cell accumulation in Broncho-Alveolar Lavage (BAL) fluids, specifically lymphocytes and eosinophils, lung inflammation, oxidative stress, airway remodelling, and pro-inflammatory cytokine levels. H&E analysis of lung tissue sections identified attenuated inflammatory cell infiltration and thickening of bronchial epithelium by DSK. PAS staining and MT staining identified decrease in OVA-induced mucus hyper secretion and peri-bronchial collagen deposition respectively, upon DSK treatment. Treatment with DSK increased the mRNA expression of antioxidative defence gene Nrf-2 and its downstream target genes HO-1 and NQO-1. In the same line, biochemical analysis for the markers of oxidative/antioxidant system confirmed the restoration of activity of Catalase, GPx, SOD and EPO and the levels of GSH, GSSG, MDA and Nitrite in whole lungs. In line with PAS staining, DSK treatment decreased the OVA-induced expression of Muc5AC and Muc5B genes. DSK treatment reduced the steady state mRNA expression levels of IL-6, IL-1β, TNF-α, IL-4, -5, -33, IFN-γ in whole lung; and IL-6, TNF-α and IL-1β protein levels in BALF.. Collectively, our results suggest that herbal decoction DSK is effective in protecting against allergic airway inflammation and remodelling by regulating anti-oxidant mechanisms. We postulate that DSK could be the potential therapeutic option for allergic asthma management.

Topics: Airway Remodeling; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; Asthma; Cytokines; Disease Models, Animal; Eosinophils; Hypersensitivity; Immunologic Factors; Lung; Male; Medicine, Ayurvedic; Mice, Inbred BALB C; NF-E2-Related Factor 2; Ovalbumin; Oxidative Stress; Plant Preparations; Pneumonia

Topics: Allergens; Animals; Asthma; Cytokines; Formaldehyde; Immunoglobulin E; Lung; Male; Mice, Inbred BALB C; Ovalbumin; Respiratory Function Tests; Temperature; TRPA1 Cation Channel; TRPM Cation Channels

Airway dendritic cells (DCs) are recognized as important factors in the mechanisms of allergic inflammatory diseases. Suppressor of cytokine signaling 3 (SOCS3) is involved in regulating the functions of T cells and macrophages, but the roles of SOCS3-expressing DCs in the pathogeneses of allergic inflammatory diseases are still controversial. We compared the effects of adoptively transferred SOCS3

Topics: Adoptive Transfer; Animals; Asthma; Bone Marrow Cells; Cell Movement; Cell Proliferation; Chemotaxis; Cytokines; Dendritic Cells; Hypersensitivity; Inflammation; Injections, Intraperitoneal; Lipopolysaccharides; Lung; Lymphocyte Culture Test, Mixed; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Signal Transduction; STAT Transcription Factors; Suppressor of Cytokine Signaling 3 Protein; T-Lymphocytes

Asthma is a chronic inflammatory disease affecting millions of people around the world, yet much remains unknown about its underlying mechanisms. Cortistatin (CST) is a neuropeptide which is reported to be a potential endogenous anti-inflammatory factor in several conditions. To testify the potential involvement of CST in airway inflammatory reaction, an ovalbumin (OVA)-induced mice model was established in wild-type (WT) as well as CST-knockout (Cort

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Asthma; Chemokine CCL2; Disease Models, Animal; Lung; Male; Mice, Knockout; Neuropeptides; NF-kappa B; Ovalbumin; Signal Transduction

BACKGROUND Loss of the epithelial barrier is characterized by a reduction in E-cadherin expression and is a hallmark of asthma. Qi-xian decoction (QXT) is a Chinese medicinal formula that has been used to effectively treat asthma. This study aimed to investigate the effect of QXT on E-cadherin expression in human lung epithelial 16HBE cells and ovalbumin-challenged mice and to explore the underlying molecular mechanism. MATERIAL AND METHODS Ovalbumin (OVA)-induced mice were used as a model of asthma. Real-time PCR and Western blotting were utilized to examine mRNA and protein levels. Lung tissue reactive oxygen species (ROS) levels were evaluated using dichloro-dihydro-fluorescein diacetate (DCFH-DA). Serum superoxide dismutase (SOD) and the total antioxidant capacity (TAOC) were measured via enzyme-linked immunosorbent assay (ELISA)-based analyses. 16HBE cells were utilized to explore the effect of QXT or hydrogen peroxide (H₂O₂) on the expression of E-cadherin in vitro. RESULTS We found that QXT treatment increased E-cadherin expression and decreased extracellular-signal-regulated kinase (ERK) phosphorylation levels in the lung tissues of OVA-challenged mice. QXT also downregulated ROS levels and increased serum SOD and TAOC levels in OVA-challenged mice. In vitro studies demonstrated that increased ROS generation induced by H₂O₂ resulted in decreased E-cadherin expression levels in 16HBE cells, which was attenuated by inhibition of ERK signaling. Moreover, the H₂O₂-induced downregulation of E-cadherin expression, increased ROS generation, and ERK activation in 16HBE cells were restored by treatment with QXT water or ethanol extract. CONCLUSIONS These data demonstrate that one mechanism by which QXT protects against asthma is to restore E-cadherin expression in vivo and in vitro by inhibiting ROS-mediated ERK activation.

Topics: Animals; Asthma; Cadherins; Disease Models, Animal; Enzyme Activation; Epithelial Cells; Extracellular Signal-Regulated MAP Kinases; Female; Humans; Hydrogen Peroxide; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphorylation; Reactive Oxygen Species; Up-Regulation

Asthma, one of the significant public health problems, is triggered by certain inflammatory processes in the airways that are not addressed propitiously by current therapies. Though pieces of evidence on allergic asthma mitigation by the anti-inflammatory bioflavonoid chrysin (CHR) are accumulating, poor bioavailability, and low solubility curtail drug development. To overcome these shortcomings, CHR loaded nanoparticle (CHR-NP) was formulated, and its salutary effect in preclinical murine allergic asthma model via the peroral route was evaluated. The spherical nanosized particles showed slow, sustained release in vitro. Moreover, CHR-NP dramatically reduced the serum IgE, ovalbumin (OVA)-induced lung histological alteration, as well as Th2 (T-helper 2) cytokines in the bronchoalveolar lavage fluid (BALF). It also suppressed the elevated serum pro-inflammatory cytokines and their upstream TLR/NF-κB/NLRP3 pathway activation in lung superior to CHR and almost identical to dexamethasone (DEX). Thus this study suggests the potentiality of CHR-NP in ameliorating allergic asthma progression.

Topics: A549 Cells; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Flavonoids; Humans; Hypersensitivity; Immunoglobulin E; Inflammation Mediators; Lung; Male; Mice; Mice, Inbred BALB C; Microscopy, Atomic Force; Microscopy, Electron, Transmission; NF-kappa B; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Polylactic Acid-Polyglycolic Acid Copolymer; Toll-Like Receptors

Asthma is a chronic, serious allergic inflammatory disease in the airway. The inflammation in the airway is induced by the allergic T-helper 2 cells (Th2 cells), which leads to unfettered production of inflammatory cytokines. The accretion of inflammatory cells in the airway also speeds up the secretion of reactive oxygen species (ROS) and suppresses antioxidative processes. Hence, the present work aimed to study the antiasthmatic efficacy of betulin and its effect in suppressing the inflammatory markers of ovalbumin (OVA) challenged asthmatic mice. The observed results revealed that the levels of inflammatory cells including neutrophils, eosinophils, lymphocytes, and macrophages were effectively decreased by betulin treatment; furthermore, the inflammatory markers IL-4, IL-5, IL-13, and TNF-α levels were notably suppressed by betulin administration in OVA-challenged asthmatic mice. Similarly, the oral administration of betulin showed a reduction in IgE level and elevation in the IFN-γ level in bronchoalveolar lavage fluid (BALF). The elevated levels of antioxidant enzymes like catalase (CAT), glutathione (GSH), and superoxide dismutase (SOD) were observed in betulin treated mice. Furthermore, reduced levels of reactive oxygen species like NO2, NO3, and MDA were noted in the betulin treated group. Consistently, airway hyperreactivity (AHR) was depleted in the betulin administered group compared with the OVA-challenged asthmatic group. Betulin treatment was revealed to have noteworthy antiasthmatic effects mediated by the suppression of production of inflammatory cells and the expression of other inflammatory markers. Furthermore, the elevation in the level of antioxidant markers helped to disclose the original regulatory mode of betulin on asthma treatment.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Female; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Reactive Oxygen Species; Respiratory Function Tests; Triterpenes

Asthma is marked by chronic irritation in the airway lumen of the lungs due to the accretion of inflammatory cells that influence the regular inhalation process. An extended buildup of inflammation leads to oxidative pressure and the repression of antioxidant functions. In the current study, a potential compound, boldine, was tested for the containment of provocative markers along the path of antiasthmatic activity in an ovalbumin (OVA)-induced asthmatic mice model. As an effect, the boldine (10 and 20 mg/kg) treatment suppressed inflammatory cells such as eosinophil, macrophage, neutrophil, lymphocyte, and other inflammatory markers in the bronchoalveolar lavage fluid (BALF) of OVA-induced mice. Likewise, immunoglobulin E (IgE) levels were drastically condensed in the serum of boldine-treated animals. Levels of enzymatic and nonenzymatic antioxidants, such as superoxide dismutase (SOD) and glutathione (GSH), were upregulated in the boldine treatment group compared to the asthmatic control group, which displays the antioxidant effects of boldine on asthmatic animals. Interestingly, the reactive oxygen species (ROS) and malonaldehyde (MDA) levels were repressed in the BALF of boldine-treated mice groups. Therefore, the effects of boldine are significant for the management of asthma, reducing the accrual of inflammatory cells, along with other inflammatory markers, while improving antioxidant markers and containing ROS. Hence, boldine may be an option for clinical trials of chronic asthma management.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antioxidants; Aporphines; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Eosinophils; Immunoglobulin E; Lung; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Reactive Oxygen Species; Respiratory Function Tests

Topics: Animals; Anti-Inflammatory Agents; Asthma; beta-N-Acetylhexosaminidases; Body Weight; Bronchi; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Capsaicin; Cordyceps; Cytokines; Disease Models, Animal; Eosinophil Peroxidase; Female; Histamine Release; Immunization; Immunoglobulin E; Mast Cells; Methacholine Chloride; Mice, Inbred BALB C; Mycelium; Nasal Lavage; Ovalbumin; Rats, Sprague-Dawley; Rhinitis, Allergic; Skin; Spleen; Trachea

Asthma is defined as a heterogeneous disease, usually characterized by chronic airway inflammation. Long noncoding RNAs (lncRNAs) play important roles in various biological processes. To know more about the relationships between lncRNAs and asthma, gene microarray analysis was performed to screen differentially expressed lncRNAs between the lung tissue of ovalbumin (OVA) mice and control mice. Further studies showed that downregulating differentially expressed lncRNA-AK149641 by adeno-associated virus 6 (AAV6) in OVA mice inhibited airway inflammation, with improved airway compliance and resistance, diminished infiltration of inflammatory cells, as well as less secretions of mucus, tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6). Moreover, the activity of nuclear factor-kappa B (NF-κB) in the lung tissue was reduced after downregulating lncRNA-AK149641. In conclusion, we proposed that downregulation of lncRNA-AK149641 attenuated the airway inflammatory response in an OVA-induced asthma mouse model, probably in association with modulation of the NF-κB signaling pathway.

Topics: Animals; Asthma; Disease Models, Animal; Female; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Long Noncoding

Pentraxin 3 (PTX3) regulates multiple aspects of innate immunity and tissue inflammation. Recently, it has been reported that PTX3 deficiency enhances interleukin (IL)-17A-dominant pulmonary inflammation in an ovalbumin (OVA)-induced mouse asthma model. However, whether PTX3 treatment would provide protection against allergic airway inflammation has not been clearly elucidated. The goal of this study was to further investigate the effect of recombinant PTX3 administration on the phenotype of asthma.. C57BL/6 J mice were sensitized and challenged with OVA to induce eosinophilic asthma model, as well as sensitized with OVA plus LPS and challenged with OVA to induce neutrophilic asthma model. We evaluated effect of recombinant PTX3 on asthma phenotype through both asthma models. The bronchoalveolar lavage fluid (BALF) inflammatory cells and cytokines, airway hyperresponsiveness, and pathological alterations of the lung tissues were assessed.. In both eosinophilic and neutrophilic asthma models, PTX3 treatment provoked airway hyperresponsiveness, concomitant with increased inflammatory cytokines IL-4, IL-17, eotaxin, and transforming growth factor (TGF)-β1 and aggravated airway accumulation of inflammatory cells, especially eosinophils and neutrophils. In histological analysis of the lung tissue, administration of PTX3 promoted inflammatory cells infiltration, mucus production, and collagen deposition. In addition, PTX3 also significantly enhanced STAT3 phosphorylation in lung tissue.. Our results show that exogenous PTX3 can exacerbate multiple asthmatic features by promoting both eosinophils and neutrophils lung infiltration and provide new evidence to better understand the complex role of PTX3 in allergic airway inflammation.

Topics: Animals; Asthma; C-Reactive Protein; Female; Inflammation Mediators; Mice; Mice, Inbred C57BL; Nerve Tissue Proteins; Ovalbumin

Topics: Animals; Anti-Inflammatory Agents; Asthma; Dexamethasone; Disease Models, Animal; Drug Carriers; Hydrogen Peroxide; Hypersensitivity; Mice; Mice, Inbred BALB C; Nanoparticles; Ovalbumin

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Dendritic Cells; Disease Models, Animal; Drugs, Chinese Herbal; Immunity; Interferon-alpha; Interferon-gamma; Interleukin-12; Male; Mice; Mice, Inbred BALB C; Ovalbumin; RNA, Messenger; Toll-Like Receptors

This study aimed to evaluate the effects of early-life exposure to different extracts of Angiostrongylus cantonensis (A. cantonensis) on airway inflammation in an allergic asthma model. The total soluble extract (TE) and the soluble extracts of the digestive (AcD), reproductive (AcR), and cuticle (AcC) systems of A. cantonensis were used for immunisation before ovalbumin (OVA)-sensitisation/challenge in an OVA-induced allergic asthma model. The initial hypothesis of the study was that some soluble extract of the systems (AcD, AcR, or AcC) could be more potent to the modulation of inflammation than the TE. Our data, however, shows that immunisation with the TE is more promising because it decreased the high influx of inflammatory cells on airways and promoted an increase of interferon-γ (IFN-ɣ) and interleukin-10 (IL-10) levels. Besides this, the immunisation with the TE also led to a reduction of goblet cells and mucus overproduction in the lung tissue of asthmatic mice. We believe that the extracts have a distinct capacity to modulate the immune system, due to the TE possessing a greater variability of molecules, which together leads to control of airway inflammation. In conclusion, this is the first study to reveal that the TE of A. cantonensis adult worms has a greater potential for developing a novel therapeutic for allergic asthma.

Topics: Angiostrongylus cantonensis; Animals; Asthma; Cytokines; Disease Models, Animal; Female; Immunization; Immunomodulation; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Mucosa

The aryl hydrocarbon receptor (AhR) is a ligand-dependent-activated transcriptional factor that regulates the metabolism of xenobiotic and endogenous compounds. Recent studies have shown that AhR is a novel master regulator of the mucosal immune system, including lungs and intestine. To elucidate the role of AhR in chronic severe asthma, AhR wild-type and knockout mice (AhR

Topics: Animals; Asthma; Basic Helix-Loop-Helix Transcription Factors; Cell Movement; Cytokines; Disease Models, Animal; Epithelial-Mesenchymal Transition; Female; Inflammation; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Receptors, Aryl Hydrocarbon; Respiratory Hypersensitivity; Th17 Cells

Asthma is a common respiratory immune disease in children and adults, and interleukin-4 (IL-4) is one of the key factors for the onset of asthma. Therefore, targeting human IL-4 and IL-4 receptor alpha (IL-4RA) has become one of the strategies for targeted therapy of cytokines. Herein, we established an animal model of asthmatic airway inflammation using double humanized IL-4/IL-4RA (hIL-4/hIL-4RA) mice, where human IL-4 and IL-4RA replaced their murine counterparts, respectively. We successfully identified the phenotype by Southern blotting, ELISA, and flow cytometry. The hIL-4/hIL-4RA mice induced by ovalbumin (OVA) exhibited several important features of asthma, such as inflammatory cell infiltration, IgE release, goblet cell hyperplasia, and Th2 cytokine secretion. Furthermore, treatment of these humanized mice with anti-human IL-4RA antibodies significantly inhibited level of these pathological indicators. Thus, hIL-4/hIL-4RA mice provide a validated preclinical mouse model to interrogate new therapeutic agents targeting this specific cytokine pathway in asthma.

Topics: Allergens; Animals; Antibodies, Monoclonal; Asthma; Bronchoalveolar Lavage Fluid; Cells, Cultured; Disease Models, Animal; Female; Gene Editing; Goblet Cells; Humans; Immunoglobulin E; Interleukin-4; Interleukin-4 Receptor alpha Subunit; Leukocytes; Lung; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Transgenic; Mucus; Ovalbumin; Spleen

Silica crystals (silica), which are a major mineral component of volcanic ash and desert dust, contribute to the pathogenesis of pulmonary disorders such as asthma and fibrosis. Although administration of silica or sand dust to rodents exacerbates development of ovalbumin-induced or house dust mite-induced asthma-like airway inflammation, the detailed mechanisms remain unclear. Here, using murine models, we found that silica can induce IL-33 expression in pulmonary epithelial cells. IL-33, but not IL-25 or TSLP, and type 2 cytokines such as IL-5 and IL-13 were critically involved in silica's exacerbation of OVA-induced airway eosinophilia in mice. Innate lymphoid cells (ILCs), but not T, B or NKT cells, were also involved in the setting. Moreover, a scavenger receptor that recognized silica was important for silica's exacerbating effect. These observations suggest that IL-33 induced in epithelial cells by silica activates ILCs to produce IL-5 and/or IL-13, contributing to silica's exacerbation of OVA-induced airway eosinophilia in mice. Our findings provide new insight into the underlying mechanisms of exacerbation of pulmonary disorders such as asthma following inhalation of silica-containing materials such as volcanic ash and desert dust.

Topics: Animals; Asthma; Cytokines; Interleukin-13; Interleukin-33; Interleukin-5; Interleukins; Lung; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Pneumonia; Pulmonary Eosinophilia; Receptors, Scavenger; Silicon Dioxide; Thymic Stromal Lymphopoietin

Asthma is a chronic respiratory disease which is susceptible to children and causes great harm to them. Recently, Interferon-related developmental regulator 1 (IFRD1) was proved to be participant in regulating lung diseases, and its abnormal expression was shown in pathological airway tissues. Our study aimed to demonstrate the role and modulatory mechanism of IFRD1 in the pathogenesis of asthma. First, we evaluated the expression of IFRD1 in the lungs of asthmatic patients. C57BL/6 mice and human bronchial epithelioid (HBE) cells were respectively induced by ovalbumin (OVA) and lipopolysaccharide (LPS) to construct asthma models in vivo and in vitro. Using adenovirus and pcDNA vectors, we carried out overexpression assays on mice and cell models. Additionally, the potential mechanism of IFRD1 on regulating asthma process was elucidated by targeting NF-κB pathway. The results showed that IFRD1 was significantly down-regulated in asthma lung tissues, as well as the in vivo and in vitro models of asthma. Besides, OVA induced the inflammation responses and hyperreactivity of airway in mice, and LPS also caused inflammatory cytokine secretion and apoptosis of HBE cells, while cell viability was inhibited. However, IFRD1 overexpression dramatically reversed the effects of OVA and LPS. We subsequently discovered that the NF-κB pathway was activated in asthmatic cells, and NF-κB signaling activation was involved in IFRD1 regulated asthma responses of HBE cells. In conclusion, our study indicated that IFRD1 inhibited the asthmatic responses of airway via the NF-κB pathway inactivation. The evidence presented herein might provide a novel sight for asthma therapy.

Topics: Animals; Apoptosis; Asthma; Cell Survival; Child; Down-Regulation; Female; Humans; Immediate-Early Proteins; Inflammation; Lipopolysaccharides; Lung; Male; Membrane Proteins; Mice, Inbred C57BL; NF-kappa B; Ovalbumin; Signal Transduction

Dryopteris crassirhizoma (DC) has a wide range of pharmacological effects, including antibacterial, anti‑influenza virus, anti‑tumor, anti‑reverse transcriptase and antioxidant effects. However, the inhibitory effect of DC on allergic inflammatory response remains unclear; therefore, the current study used an experimental ovalbumin (OVA)‑induced allergic asthma mouse model and phorbol myristate acetate (PMA)‑ and A23187‑stimulated HMC‑1 cells to reveal the effects of DC in regulating airway inflammation and its possible mechanism. Allergic asthma was initiated in BALB/c mice via exposure to OVA emulsified in aluminum, on days 1 and 14. Thereafter, the mice were treated with DC or dexamethasone (Dex) orally, before being challenged, from days 15 to 26. Subsequently, the mice were challenged with OVA on days 27, 28 and 29. The results of histological analysis indicated that the administration of DC decreased the number of inflammatory cells in the bronchoalveolar lavage fluid (BALF) and suppressed eosinophilic infiltration, mucus production and collagen deposition in the lung tissue. DC treatment increased the level of T helper type 1 (Th1) cytokines (IL‑10 and interferon (IFN)‑γ) and decreased the levels Th2 cytokines (IL‑4, IL‑5 and IL‑13) and proinflammatory cytokines (IL‑6 and TNF‑α). Furthermore, DC treatment inhibited the activation of NF‑κB signaling (NF‑κB, p‑NF‑κB, IκB and p‑IκB), both in BALF and lung homogenates. Serum levels of total IgE and OVA‑specific IgE and IgG1 were significantly lower after DC treatment compared with after OVA treatment. However, the anti‑inflammatory effect of OVA‑specific IgG2a was higher after DC treatment. In addition, DC treatment attenuated the production of proinflammatory cytokines, including IL‑6 and TNF‑α, and the activation of NF‑κB signaling (NF‑κB and p‑NF‑κB), in PMA and calcium ionophore A23187‑stimulated HMC‑1 cells. In summary, the current study demonstrated that DC acts a potent anti‑allergic and anti‑inflammatory drug by modulating the Th1 and Th2 response and reducing the allergic inflammatory reaction in PMA and A23187‑stimulated HMC‑1 cells via NF‑κB signaling in an OVA‑induced allergic asthma model.

Topics: Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Calcimycin; Cell Line, Tumor; Cytokines; Disease Models, Animal; Dryopteris; Humans; Lung; Male; Mast Cells; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Phytotherapy; Plant Extracts; Signal Transduction; Tetradecanoylphorbol Acetate

Mounting evidence, consistent with our previous study, showed that γ-aminobutyric acid type A receptor (GABAAR) played an indispensable role in airway inflammation and mucus hypersecretion in asthma. Monocyte chemotactic protein-inducing protein 1 (MCPIP1) was a key negative regulator of inflammation. Recent studies showed that inflammation was largely suppressed by enhanced MCPIP1 expression in many inflammatory diseases. However, the role and potential mechanism of MCPIP1 in airway inflammation and mucus hypersecretion in asthma were still not well studied. This study was to explore the role of MCPIP1 in asthmatic airway inflammation and mucus hypersecretion in both mice and BEAS-2B cells, and its potential mechanism.. In vivo, mice were sensitized and challenged by ovalbumin (OVA) to induce asthma. Airway inflammation and mucus secretion were analyzed. In vitro, BEAS-2B cells were chosen. Interleukin (IL)-13 was used to stimulate inflammation and mucus hypersecretion in cells. MCPIP1 Lentiviral vector (LA-MCPIP1) and plasmid-MCPIP1 were used to up-regulate MCPIP1 in lung and cells, respectively. MCP-1, thymic stromal lymphopoietin (TSLP), mucin 5AC (MUC5AC), MCPIP1, and GABAARβ2 expressions were measured in both lung and BEAS-2B cells. Immunofluorescence staining was performed to observe the expression of GABAARβ2 in cells.. MCPIP1 was up-regulated by LA-MCPIP1 (P < 0.001) and plasmid-MCPIP1 (P < 0.001) in lung and cells, respectively. OVA-induced airway inflammation and mucus hypersecretion, OVA-enhanced MCP-1, TSLP, MUC5AC, and GABAARβ2 expressions, and OVA-reduced MCPIP1 were significantly blunted by LA-MCPIP1 in mice (all P < 0.001). IL-13-enhanced MCP-1, TSLP, MUC5AC, and GABAARβ2 expressions, and IL-13-reduced MCPIP1 were markedly abrogated by plasmid-MCPIP1 in BEAS-2B cells (all P < 0.001).. The results of this study suggested that OVA and IL-13-induced airway inflammation and mucus hypersecretion were negatively regulated by MCPIP1 in both lung and BEAS-2B cells, involving GABAAR signaling pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; gamma-Aminobutyric Acid; Inflammation; Mice; Mice, Inbred BALB C; Monocytes; Mucus; Ovalbumin; Ribonucleases; Signal Transduction

Study found that glucocorticoids, as first-line treatments for asthma, fails to prevent asthma recurrence. Orosomucoid-like (ORMDL) 3 is associated to childhood asthma onset and involved in the inflammation and repair of airway epithelium. We explored the functional role of ORMDL3 in glucocorticoid treatment for childhood asthma.. Mice were sensitized with Ovalbumin (OVA) and treated with Dexamethasone (Dex), followed by OVA challenge to establish a mouse model of asthma. Histopathological changes in lung tissues were observed by hematoxylin-eosin and masson staining. Human bronchial epithelial (16HBE-14°) cells were transfected with ORMDL3 overexpression plasmid and siRNA-interleukin (IL)-33 alone or in combination, followed by Dex. Cell viability was measured by MTT assay. Cell migration was evaluated by wound healing assay. The expressions of E-cadherin and Vimentin and the activation of NF-κB and MAPK/ERK in 16HBE-14° cells were assessed by Western blot. The expressions of ORMDL3 and IL-33 in lung tissues and 16HBE-14° cells were analyzed by qRT-PCR or Western blot.. Dex treatment alleviated the histopathological abnormality and reversed the overexpressions of ORMDL3 and IL-33 in the lung tissues of asthmatic mice. Overexpressed ORMDL3 enhanced migration and viability, decreased E-cadherin level, increased the levels of IL-33 and Vimentin, and promoted the phosphorylation of NF-κB and MAPK/ERK in Dex-treated 16HBE-14° cells, thus reversing the effect of Dex treatment. However, siRNA-IL-33 inhibited viability and migration, increased E-cadherin level, decreased Vimentin level, and suppressed the phosphorylation of NF-κB and MAPK/ERK, thus reversing the effect of overexpressed ORMDL3 in Dex-treated 16HBE-14° cells.. ORMDL3 overexpression helped airway epithelial cellrepairin asthma via regulating IL-33 expression.

Topics: Animals; Asthma; Disease Models, Animal; Epithelial Cells; Glucocorticoids; Interleukin-33; Lung; Membrane Proteins; Mice; Mice, Inbred BALB C; Ovalbumin

Allergic asthma is the most common type of asthma which characterized by inflammatory responses of the airways. Alpinetin, a flavonoid compound derived from the ginger family of medicinal herbs, possesses various biological properties including anti-inflammatory, anti-oxidant and other medical effects. In this study, we aimed to evaluate the effects of alpinetin on OVA-induced allergic asthma, and further to examine its molecular mechanisms underlying these processes in vivo and in vitro. Mice were sensitized and challenged with OVA to build allergic asthma model in vivo. Bronchoalveolar lavage fluid (BALF) was collected for inflammatory cells analysis and lung tissues were examined for histopathological examination. The levels of IL-5, IL-13, IL-4, IgE, TNF-α, IL-6 and IL-1β were determined by the respective ELISA kits. The PI3K/AKT/NF-κB and HO-1 signaling pathways were examined by western blot analysis. The results showed that alpinetin significantly ameliorated OVA-induced pathologic changes of lungs, such as decreasing massive inflammatory cell infiltration and mucus hypersecretion, and reduced the number of inflammatory cells in BALF. Alpinetin also decreased the OVA-induced levels of IL-4, IL-5, IL-13 and IgE. Furthermore, alpinetin inhibited OVA-induced phosphorylation of p65, IκB, PI3K and AKT, and the activity of HO-1 in vivo. More importantly, these anti-inflammatory effects and molecular mechanisms of alpinetin has also been confirmed in LPS-stimulated RAW 264.7 macrophages in vitro. In conclusion, above results indicate that alpinetin exhibites a potent anti-inflammatory activity in allergic asthma through modulating PI3K/AKT/NF-κB and HO-1 signaling pathways, which would be used as a promising therapy agent for allergic asthma.

Topics: Animals; Asthma; Cell Survival; Drug Tapering; Flavanones; Gene Expression Regulation; Heme Oxygenase-1; Immunoglobulin E; Inflammation; Interleukin-4; Membrane Proteins; Mice; Molecular Structure; NF-kappa B; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; RAW 264.7 Cells; Signal Transduction

Allergic airway inflammation is one of the major pathological events involved in the development of asthma. The B cell-activating factor (BAFF)-mediated abnormal activation of B cells plays a key role in developing allergic airway inflammation. Here, we investigated the effects of Gu-Ben-Fang-Xiao decoction (GBFXD), a TCM decoction used in the prevention and treatment of allergic asthma, on allergic airway inflammation and BAFF-mediated B cell activation. A mouse model of OVA-Severe respiratory syncytial virus (RSV) induced asthma in the remission stage was administrated with GBFXD by gavage for four weeks, after which, the pulmonary function was evaluated. Pathological changes of the lung were observed by hematoxylin and eosin (HE) staining, and serum levels of IgE, BAFF, and inflammatory factors were detected by ELISA. The expression of BAFF, APRIL, and their related receptors in the lung and spleen was detected by Western blotting and RT-qPCR. Flow cytometry detected B cell subsets in the spleen, PBC, and monocyte subsets in bronchoalveolar lavage fluid (BALF). The results showed that GBFXD improved the lung function, alleviated the inflammatory changes of the lung tissue in OVA-RSV sensitized mice, and reduced levels of IL-6, TNF-α, IL1-β, INOS, IL13 as well as IL-15, IgE, BAFF in the serum of OVA-RAV mice. Additionally, GBFXD significantly reduced the proportion of CD19+CD27+ B cell subpopulation and IgE + B cell subpopulation in the PBC and spleen cells of mice. Furthermore, the expression of BAFF, APRIL, BAFFR, TACI, and AID decreased in the lung and spleen of GBFXD-treated mice, as well as the proportion of CD11b + BAFF + cell subsets in BALF. In conclusion, GBFXD has an inhibitory effect on the secretion of BAFF by pulmonary macrophages and the expression of BAFF-related receptors, thereby reducing B cell activation and the release of IgE. This proposed mechanism contributes to the improvement of allergic airway inflammation and respiratory function in an asthmatic mouse model.

Topics: Animals; Asthma; B-Cell Activating Factor; B-Lymphocytes; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Drugs, Chinese Herbal; Female; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Syncytial Viruses

Topics: Animals; Apoptosis; Asthma; Dendritic Cells; Disease Models, Animal; Dose-Response Relationship, Immunologic; Gene Expression Regulation; Glucocorticoid-Induced TNFR-Related Protein; Humans; Immune Tolerance; Lipopolysaccharides; Lymphocyte Count; Mice; Mice, Inbred BALB C; Ovalbumin; Signal Transduction; Specific Pathogen-Free Organisms; T-Lymphocytes, Regulatory; Th17 Cells; Th2 Cells; Toll-Like Receptor 4; Tumor Necrosis Factors

In allergic bronchial asthma, an increased smooth muscle contractility of the airways is one of the causes of the airway hyperresponsiveness (AHR). Increasing evidence also suggests a possible involvement of microRNAs (miRNAs) in airway diseases, including asthma, although their roles in function and pathology largely unknown. The current study aimed to determine the role of a miRNA, miR-140-3p, in the control of protein expression of CD38, which is believed to regulate the contraction of smooth muscles, including the airways. In bronchial smooth muscles (BSMs) of the mice that were actively sensitized and repeatedly challenged with ovalbumin antigen, an upregulation of CD38 protein concurrently with a significant reduction of miR-140-3p was observed. In cultured human BSM cells (hBSMCs), transfection with a synthetic miR-140-3p inhibitor caused an increase in CD38 protein, indicating that its basal protein expression is regulated by endogenous miR-140-3p. Treatment of the hBSMCs with interleukin-13 (IL-13), an asthma-related cytokine, caused both an upregulation of CD38 protein and a downregulation of miR-140-3p. Transfection of the hBSMCs with miR-140-3p mimic inhibited the CD38 protein upregulation induced by IL-13. On the other hand, neither a CD38 product cyclic ADP-ribose (cADPR) nor its antagonist 8-bromo-cADPR had an effect on the BSM contraction even in the antigen-challenged mice. Taken together, the current findings suggest that the downregulation of miR-140-3p induced by IL-13 might cause an upregulation of CD38 protein in BSM cells of the disease, although functional and pathological roles of the upregulated CD38 are still unclear.

Topics: ADP-ribosyl Cyclase 1; Animals; Asthma; Bronchi; Cell Line; Disease Models, Animal; Gene Expression Regulation; Humans; Hypersensitivity; Interleukin-13; Male; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; MicroRNAs; Myocytes, Smooth Muscle; Ovalbumin

Increased IL-8 levels and neutrophil accumulation in the airways are common features found in patients affected by pulmonary diseases such as Asthma, Idiopathic Pulmonary Fibrosis, Influenza-A infection and COPD. Chronic neutrophilic inflammation is usually corticosteroid insensitive and may be relevant in the progression of those diseases.. To explore the role of Ladarixin, a dual CXCR1/2 antagonist, in several mouse models of airway inflammation with a significant neutrophilic component.. Ladarixin was able to reduce the acute and chronic neutrophilic influx, also attenuating the Th2 eosinophil-dominated airway inflammation, tissue remodeling and airway hyperresponsiveness. Correspondingly, Ladarixin decreased bleomycin-induced neutrophilic inflammation and collagen deposition, as well as attenuated the corticosteroid resistant Th17 neutrophil-dominated airway inflammation and hyperresponsiveness, restoring corticosteroid sensitivity. Finally, Ladarixin reduced neutrophilic airway inflammation during cigarette smoke-induced corticosteroid resistant exacerbation of Influenza-A infection, improving lung function and mice survival.. CXCR1/2 antagonist Ladarixin offers a new strategy for therapeutic treatment of acute and chronic neutrophilic airway inflammation, even in the context of corticosteroid-insensitivity.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Biomarkers; Biopsy; Bleomycin; Cytokines; Disease Models, Animal; Disease Progression; Disease Susceptibility; Eosinophils; Female; Fibrosis; Immunohistochemistry; Leukocytes; Male; Mice; Mice, Knockout; Neutrophils; Ovalbumin; Oxidation-Reduction; Receptors, Interleukin-8A; Receptors, Interleukin-8B; Respiratory Hypersensitivity; Respiratory Tract Diseases; Sulfonamides; T-Lymphocyte Subsets

This study is to investigate the inhibitory effects and mechanisms of DEK-targeting aptamer (DTA-64) on epithelial mesenchymaltransition (EMT)-mediated airway remodelling in mice and human bronchial epithelial cell line BEAS-2B. In the ovalbumin (OVA)-induced asthmatic mice, DTA-64 significantly reduced the infiltration of eosinophils and neutrophils in lung tissue, attenuated the airway resistance and the proliferation of goblet cells. In addition, DTA-64 reduced collagen deposition, transforming growth factor 1 (TGF-β1) level in BALF and IgE levels in serum, balanced Th1/Th2/Th17 ratio, and decreased mesenchymal proteins (vimentin and α-SMA), as well as weekend matrix metalloproteinases (MMP-2 and MMP-9) and NF-κB p65 activity. In the in vitro experiments, we used TGF-β1 to induce EMT in the human epithelial cell line BEAS-2B. DEK overexpression (ovDEK) or silencing (shDEK) up-regulated or down-regulated TGF-β1 expression, respectively, on the contrary, TGF-β1 exposure had no effect on DEK expression. Furthermore, ovDEK and TGF-β1 synergistically promoted EMT, whereas shDEK significantly reduced mesenchymal markers and increased epithelial markers, thus inhibiting EMT. Additionally, shDEK inhibited key proteins in TGF-β1-mediated signalling pathways, including Smad2/3, Smad4, p38 MAPK, ERK1/2, JNK and PI3K/AKT/mTOR. In conclusion, the effects of DTA-64 against EMT of asthmatic mice and BEAS-2B might partially be achieved through suppressing TGF-β1/Smad, MAPK and PI3K signalling pathways. DTA-64 may be a new therapeutic option for the management of airway remodelling in asthma patients.

Topics: Alveolar Epithelial Cells; Animals; Aptamers, Nucleotide; Asthma; Basic Helix-Loop-Helix Transcription Factors; Biomarkers; Chromosomal Proteins, Non-Histone; Disease Susceptibility; Epithelial-Mesenchymal Transition; Female; Gene Silencing; Humans; Immunoglobulin E; Immunomodulation; Lung; Mice; Mitogen-Activated Protein Kinases; NF-kappa B; Oncogene Proteins; Ovalbumin; Phosphatidylinositol 3-Kinases; Poly-ADP-Ribose Binding Proteins; Receptors, Aryl Hydrocarbon; Signal Transduction; Smad Proteins; T-Lymphocyte Subsets; Transforming Growth Factor beta1

Acrylamide is a toxic chemical substance produced when starch-rich foods are fried at high temperatures. Asthma is a chronic and complicated respiratory disease, of which genetic and environmental factors are the main triggers. Orally-received components may have an effect on asthma pathophysiology. The aim of this study was to investigate the role of AA as a stimulus in asthma. BALB/c mice were allocated into four groups as follows: two OVA-sensitized asthmatic groups, including one treated with AA by gavage feeding and one non-treated (asthma group), and two healthy (non-asthmatic) groups, one treated with AA by gavage feeding and one non-treated (negative control group). Airway hyperresponsiveness, cell count, cytokine levels in BAL fluid, lung histopathology, IgE levels, and oxidative stress indices including plasma level of MDA, pulmonary antioxidant enzymes (SOD and CAT) levels, HP content, and collagen fiber accumulation in lung tissue were measured. We found that the group of mice treated with both OVA and AA (asthmatic and AA-treated mice) experienced higher levels of asthma-associated biomarkers, including higher enhanced pause (Penh value), eosinophilic inflammation, mucus hyper secretion, goblet cell hyperplasia, total and OVA-specific IgE levels, IL-4, IL-5, and IL-13 levels than the group sensitized only with OVA (asthmatic mice). The OVA-AA-treated mice also experienced worsened levels of oxidative stress indicators. Healthy (non-asthmatic) mice that only received AA were in similar conditions to healthy untreated mice (negative control group). The OVA-AA-treated group showed more severe allergic asthma symptoms in comparison to the group only sensitized with OVA. Therefore, food/water contaminated with AA can act as a stimulant of allergic asthma and exacerbate the bronchial inflammatory responses.

Topics: Acrylamide; Administration, Oral; Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Cytokines; Female; Fibrosis; Inflammation Mediators; Lung; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Respiratory Hypersensitivity

Asthma is a leading allergic disease worldwide, demonstrating an ever‑increasing prevalence over the past two decades. Asthma is characterized by allergen‑associated airway hyperresponsiveness (AHR) that primarily results from T helper 2 (Th2) cell inflammation, in which dendritic cells (DCs) serve an important role in determining T cell development after encountering an antigen. Atractylodin (ATL), a polyethene alkyne extracted from Atractylodis rhizoma (also known as Cangzhu), has proven effective in treating digestive disorders, rheumatic disease and influenza. In addition, ATL was discovered to alleviate mouse collagen‑induced arthritis via regulating DC maturation. The present study aimed to investigate the effect of ATL on asthma given that DCs serve an essential role in Th2‑mediated inflammation in asthma. Mouse model of asthma was induced by ovalbumin (OVA). OVA‑induced airway hyperresponsiveness (AHR) and inflammatory cells in bronchoalveolar lavage fluid (BALF) were detected. The production of IgE and IgG1 in serum and cytokines in BALF were detected by ELISA. The effects of ATL on dendritic cells maturation and T cell expansion were detected by flow cytometry analysis and 3H‑thymidine incorporation. Using a model of OVA‑induced asthma, it was demonstrated that ATL ameliorated AHR and decreased the levels of IL‑4, IL‑5 and IL‑13 in bronchoalveolar lavage fluid (BALF), and OVA‑specific IgE and IgG1 in the serum. OVA‑stimulated splenocytes were used to demonstrated that ATL decreased cell expansion and the production of IL‑4, IL‑5 and IL‑13 in the culture medium. In order to determine the cellular mechanism of ATL in asthma, splenic DCs were isolated and it was subsequently observed that ATL downregulated the expression levels of CD40 and CD80. Furthermore, OVA‑stimulated CD4+ T cells were co‑cultured with splenic DCs, which revealed that ATL‑treated splenic DCs led to impaired cellular proliferation and the production of IL‑4, IL‑5 and IL‑13 in OVA‑stimulated T cells. In conclusion, these results indicated that ATL may suppress antigen‑specific Th2 responses in an OVA‑induced allergic asthma model via regulating DCs. Therefore, ATL may exhibit therapeutic potential in the management of asthma and other allergic diseases presenting with Th2 inflammation.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; China; Cytokines; Dendritic Cells; Disease Models, Animal; Furans; Immunoglobulin E; Immunologic Factors; Inflammation; Interleukin-13; Interleukin-4; Interleukin-5; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Th2 Cells

3,4,5-Trihydroxycinnamic acid (THCA) has been reported to possess anti-inflammatory activity. However, the effect of THCA for treating allergic asthma was unknown. Therefore, in the present study, the anti-asthmatic effects of THCA were studied in both in vitro and in vivo studies. In phorbol 12-myristate 13-acetate (PMA)-stimulated A549 airway epithelial cells, THCA pretreatment decreased the mRNA expression and secretion of interleukin (IL)-8, monocyte chemoattractant protein-1 (MCP-1), and intercellular adhesion molecules 1 (ICAM-1), and reduced the mRNA expression of matrix metalloproteinase 9 (MMP-9). THCA also inhibited PMA-induced protein kinase B (AKT), mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) activation in A549 cells. In lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages, THCA pretreatment suppressed the mRNA expression of ICAM-1 and MMP-9. In addition, THCA suppressed the adhesion of EOL and A549 cells. In ovalbumin (OVA)-administered asthmatic mice, THCA exerted inhibitory activity on IL-5, IL-13, and MCP-1 in bronchoalveolar lavage fluid (BALF) and on OVA-specific immunoglobulin E (IgE) in serum. THCA attenuated the numbers of inflammatory cells in BALF and the influx of inflammatory cell in lung tissues. Furthermore, THCA downregulated the levels of inducible nitric oxide (iNOS), cyclooxygenase-2 (COX-2), and leukotriene B4 (LTB4) expression, mucus production and CREB phosphorylation as well as Penh value. These effects were accompanied by suppression of AKT, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and NF-κB activation. Therefore, the results of the current study suggest that THCA may be a valuable adjuvant or therapeutic in the prevention or treatment of allergic asthma.

Topics: Animals; Asthma; Cell Adhesion; Cell Adhesion Molecules; Cell Line, Tumor; Chemokines; Cinnamates; Female; Gene Expression Regulation; Humans; Macrophages; Mice; Mice, Inbred BALB C; Ovalbumin; Random Allocation; RAW 264.7 Cells

The aim of this study was to analyze the 4-carvomenthenol (carvo) oral treatment on the experimental model of the combined allergic rhinitis and asthma syndrome (CARAS). BALB/c mice were OVA-sensitized on day zero and 7th (50 μg/mL OVA in 10 mg/mL Al (OH)3) and OVA-challenged (5 mg/mL, 20 μL/animal) for three weeks. In the last week, the animals were dally challenged with aerosol of OVA and the carvo treatment (12.5, 25 or 50 mg/kg) occurred one hour before each OVA-challenge. Data were analyzed and p < 0.05 was considered significant. Carvo (12.5-50 mg/kg) decreased significantly the eosinophil migration into the nasal (NALF) and bronchoalveolar (BALF) cavities as well as on the nasal and lung tissues of sick animals. The treatment also decreased mucus production on both tissue sections stained with PAS (periodic acid-Schiff satin). In addition, the histological analyzes demonstrated that sick mice presented hyperplasia and hypertrophy of the lung smooth muscle layer followed by increasing of extracellular matrix and carvo (50 mg/kg) inhibited these asthmatic parameters. We analyzed the allergic rhinitis signals as nasal frictions and sneezing and observed that carvo decreased these two signals as well as serum OVA-specific IgE titer, type 2 cytokine synthesis, mainly IL-13, with increasing of IL-10 production. Decreasing of IL-13 production corroborated with decreasing of mucus production and these effects were dependent on p38MAPK/NF-κB(p65) signaling pathway inhibition. Therefore, these data demonstrated that a monoterpene of essential oils presents anti-allergic property on an experimental model of CARAS suggesting a new drug prototype to treat this allergic syndrome.

Topics: Allergens; Animals; Anti-Allergic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; Interleukin-13; Lung; Menthol; Mice, Inbred BALB C; Mucus; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Rhinitis, Allergic; Signal Transduction; Syndrome

Asthma is a chronic pulmonary inflammatory disease characterized by excessive infiltration of leukocytes into the respiratory tract. We explored the underlying mechanisms of mesenchymal stem cells (MSCs) in the treatment of allergic asthma using a rat model.. The rats were sensitized with ovalbumin (OVA) and an aluminium hydroxide emulsion, which were injected intraperitoneally, and then the sensitized rats were challenged with aerosolized OVA. Before the allergen challenge, the model rats were injected with MSCs and MSC-derived exosomes. At the same time, 2 out of the 6 groups of rats were injected with BML-284, a Wnt agonist. The degree of airway inflammation was determined by bronchoalveolar lavage fluid (BALF) and haematoxylin and eosin (H&E) staining; the degree of airway remodelling was assessed by Masson staining; Western blotting (WB) and real-time polymerase chain reaction (PCR) were performed to evaluate Wnt/β-catenin signalling pathway-related factors and the expression of epithelial-mesenchymal transition (EMT)-related proteins in lung tissues.. We showed that among the rats that were sensitized and challenged with OVA, the injection of MSCs and MSC-derived exosomes significantly reduced the total number of cells and the number of immune cells in BALF, proliferation of goblet cells and collagen deposition. Moreover, the number of BALF cells and collagen deposition increased significantly after the injection of BML-284. WB and real-time PCR showed that MSCs and MSC-derived exosomes significantly inhibited airway remodelling and EMT by restricting the Wnt/β-catenin signalling pathway, while additional injection of BML-284 suppressed the effects of MSCs and their exosomes, increased the EMT of the airway epithelium and exacerbated airway remodelling.. MSCs inhibit chronic allergic inflammation of the airway and reduce airway remodelling and EMT of the airway epithelium in the lungs of asthmatic rats. This process is partly attributed to the inhibition of the Wnt/β-catenin signalling pathway by MSC-derived exosomes.

Topics: Airway Remodeling; Animals; Asthma; beta Catenin; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Exosomes; Inflammation; Male; Mesenchymal Stem Cells; Ovalbumin; Rats; Rats, Sprague-Dawley; Wnt Signaling Pathway

Isoliquiritigenin (ILQ), an important component of Anti-Asthma Herbal Medicine Intervention (ASHMI), had shown potent anti-asthma effect in vitro in our previous study. However, poor solubility and low bioavailability hindered in vivo application to treat asthma. This study was to develop a novel ILQ loaded self-nanoemulsifying drug delivery system (ILQ-SMEDDS) with enhanced bioavailability.. The optimized SMEDDS formulation was composed of ethyl oleate (oil phase), Tween 80 (surfactant) and PEG400 (co-surfactant) at a mass ratio of 3:6:1. The physiochemical properties of ILQ-SMEDDS, including drug content, globule size, zeta potential, scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, were characterized. And the in vitro release profile, in situ intestinal absorption, in vivo pharmacokinetic parameters and the anti-asthma effect of ILQ suspension and ILQ-SMEDDS were evaluated.. The ILQ-SMEDDS had an average globule size of 20.63 ± 1.95 nm with a polydispersity index (PDI) of 0.11 ± 0.03, and its zeta potential was -12.64 ± 2.12 mV. The cumulative release rate of ILQ from ILQ-SMEDDS to the simulated gastrointestinal tract was significantly higher than that of free ILQ suspension. And area under curve with ILQ-SMEDDS was found to be 3.95 times higher than that of ILQ suspension indicating improved bioavailability by SMEDDS. Although ILQ-SMEDDS showed a slight less effective inhibitory effect on eotaxin-1 in human lung fibroblast (HFL-1) cells than free ILQ, in an ovalbumin-induced asthma model, ILQ-SMEDDS exhibited more efficacy than ILQ suspension in improving asthma-associated inflammation, including eosinophil production, ovalbumin-specific immunoglobulin E (OVA-sIgE), interleukin 4 (IL 4), interleukin 5 (IL 5) and interferon-γ (IFN-γ). Even the low dose of ILQ-SMEDDS group (10 mg/kg) showed better anti-asthma effect than that of the ILQ suspension group (20 mg/kg).. Compared with ILQ suspension, ILQ-SMEDDS showed significantly improved bioavailability and anti-asthma effect, revealing its potential as a favorable pharmaceutical agent for treating asthma.

Topics: Administration, Oral; Animals; Asthma; Biological Availability; Chalcones; Drug Carriers; Emulsions; Humans; Intestinal Absorption; Male; Nanostructures; Ovalbumin; Polyethylene Glycols; Polysorbates; Solubility; Surface-Active Agents

Topics: Animals; Asthma; Disease Models, Animal; Female; Glutaredoxins; Humans; Inflammation; Macrophage Activation; Macrophages; Mice; Mice, Inbred BALB C; Nitric Oxide; Ovalbumin; Oxidation-Reduction; Protective Agents; RAW 264.7 Cells; Recombinant Proteins; Signal Transduction; Thioredoxins

Thymocyte-expressed, positive selection-associated 1 (Tespa1) is a critical signaling molecule in thymocyte development. This study aimed to investigate the regulatory effect of Tespa1 on mast cells in the pathogenesis of asthma and its relationship with the interleukin (IL)-4/signal transducers and activators of transcription 6 (STAT6) signaling pathway.. Compared with the healthy controls, Tespa1 expression was decreased, and IgE levels were elevated in the sputum of asthmatic patients. Animal experiments showed that Tespa1. Altogether, our results indicate that Tespa1 can negatively regulate mast cell activity, and this event is related to the mast cell IL-4/STAT6 signaling pathway and could be therapeutically exploited to treat asthma attacks.

Topics: Adaptor Proteins, Signal Transducing; Animals; Asthma; Disease Models, Animal; Humans; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; STAT6 Transcription Factor; Thymocytes

There is an increasing interest in natural products and their derivatives with therapeutic benefits and less side effects compared to steroid therapy. Benzofuran derivatives display biological effects including anti-inflammatory effects. The present study aims to investigate whether (3-(7-methoxy-2-p-tolyl benzofuran-5-yl) propan-1-ol) (DK-1108), new synthetic benzofuran compound exerts anti-asthmatic effects in vitro and in vivo. DK-1108 strongly reduced the production of inflammatory mediators, cytokines and chemokines in RAW264.7 and A549 cells. DK-1108 significantly regulated the levels of AKT/MAPKs/c-Jun activation, AP-1 luciferase activity and ICAM-1 expression. Furthermore, DK-1108 effectively suppressed the adhesion of A549 and EOL-1 cells. In OVA-induced asthmatic mice, DK-1108 decreased the levels of IL-5/IL-13/IgE production, eosinophils/macrophages influx, ICAM-1/MCP-1 expression, mucus secretion and airway hyperresponsiveness (AHR). These effects of DK-1108 were accompanied by downregulation of MAPKs activation. Therefore, we suggest that DK-1108 exerts protective effect against airway inflammation and mucus overproduction, and therefore could be valuable therapeutic agent for treatment in asthma.

Topics: A549 Cells; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Benzofurans; Dose-Response Relationship, Drug; Female; Humans; Inflammation Mediators; Mice; Mice, Inbred BALB C; Ovalbumin; RAW 264.7 Cells; Respiratory Hypersensitivity; Tetradecanoylphorbol Acetate

PM2.5 causes abnormal immune response and asthma in animals. In this study, a Balb/c mouse animal model was exposed to PM2.5 to induce asthma. Lactobacillus paracasei HB89 was fed at the same time, in order to observe whether L. paracasei HB89 mitigates respiratory tract allergies stimulated by PM2.5. The results showed that PM2.5 stimulated a significant increase in white blood cells and immunoglobulin (IgE) in OVA-induced allergic Balb/c mice, and IgE in the blood further triggered the release of histamine in the lung immune cells. This not only increased overall immune cell counts, but the lymphocyte counts also increased significantly, resulting in significant inhibitions of cytokines INF-r and TGF-β, and induction of IL-4, IL-5, IL-13 and IL-17a. After feeding with HB89, apart from the absence of observable changes in body weight, the total white blood cell count in the animal blood and IgE response were also be reduced; the proliferation of immune cells in the lungs caused by PM2.5 was slowed down; and histamine and cytokines INF-r and TGF-β were secreted in large quantities, but IL- 4, IL-5, IL-13, IL-17a were inhibited, which effectively reduced the possibility of asthma induction.

Topics: Animals; Asthma; Cytokines; Disease Models, Animal; Down-Regulation; Female; Histamine; Immunoglobulin E; Lacticaseibacillus paracasei; Lymphocyte Count; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Treatment Outcome

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Differentiation; Disease Models, Animal; Female; Humans; Interferon-gamma; Interleukin-17; Lung; Lymphocyte Activation; Mechanistic Target of Rapamycin Complex 1; Mice; Mice, Transgenic; Neutrophils; Ovalbumin; Receptors, Antigen, T-Cell; Signal Transduction; Sirolimus; Th17 Cells

We established a mouse model of allergic asthma by sensitizing with chicken ovalbumin. The volatile oils and decoctions from raw, wine- and vinegar-steamed Schisandra chinensis fruits were intragastrically administrated to the mice. Atomization, serum IgE, IL-2, IL-4 and IFN-γ in the lung homogenates and pathological sections were evaluated to compare the effect of these volatile oils and decoctions on allergic asthma in mice. The results showed that all Schisandra volatile oils could significantly suppress allergic asthma in mice. Raw Schisandra volatile oil was most effective followed by volatile oils extracted from wine-steamed and vinegar-steamed Schisandra. The decoctions had no significant impact. Our findings demonstrated that volatile oil was the active ingredient in Schisandra, and raw Schisandra could be used to prevent cough and asthma.

Topics: Animals; Asthma; Cyclooctanes; Fruit; Hypersensitivity; Lignans; Male; Mice; Oils, Volatile; Ovalbumin; Plant Extracts; Polycyclic Compounds; Schisandra

The use of phytochemical plays a major role in recent therapeutic regimens. Amongst, Capillarisin (CPS), an active chemical constituent of Artemisia capillaris was found to exert anti-inflammatory and antioxidant properties. However, the protective role of CPS has not been identified against neonatal asthma. Hence, in the present study, Wistar rats were used consisting of four groups such as control, asthma-induced, CPS-pretreated asthma animals, and CPS control. At the end of the experimental period, histology of the lungs, inflammatory cell counts in bronchoalveolar lavage fluid (BALF), inflammatory markers such as interleukin (IL) -6, IL-5, IL-4, and IL-13 were measured. Results demonstrated a significant restoration in alveolar thickening and reduced goblet cell hyperplasia with suppressed inflammatory cells. Moreover, a significant reduction in leukocyte infiltration in BALF lessened hyper responsiveness, and serum IgE levels of CPS treated group. Furthermore, the CPS administration alleviated the expression levels of IL-6, IL-17, IL-4 and IL-13 compared to the asthma-induced group. To an extent, the study elicited the extra cellular matrix protein expression in the asthma-induced animals, and the results demonstrated a profound reduction in the fibrotic markers was evidenced in CPS treated animals. Thus, the results of the present investigation propose that capillarisin can be a new medicine target for the treatment of asthma-mediated complications.

Topics: Animals; Animals, Newborn; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Artemisia; Asthma; Bronchoalveolar Lavage Fluid; Chromones; Cytokines; Disease Models, Animal; Inflammation; Lung; Ovalbumin; Rats; Rats, Wistar

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Flavonoids; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

Topics: Acute Disease; Administration, Inhalation; Administration, Oral; Airway Remodeling; Airway Resistance; Animals; Anti-Asthmatic Agents; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Bronchoconstrictor Agents; Dexamethasone; Disease Models, Animal; Drug Evaluation, Preclinical; Female; Glucocorticoids; Humans; Indoles; Inflammation Mediators; Methacholine Chloride; Mice; Ovalbumin

Studies have shown autophagy participation in the immunopathology of inflammatory diseases. However, autophagy role in asthma and in eosinophil extracellular traps (EETs) release is poorly understood. Here, we attempted to investigate the autophagy involvement in EETs release and in lung inflammation in an experimental asthma model. Mice were sensitized with ovalbumin (OVA), followed by OVA challenge. Before the challenge with OVA, mice were treated with an autophagy inhibitor, 3-methyladenine (3-MA). We showed that 3-MA treatment decreases the number of eosinophils, eosinophil peroxidase (EPO) activity, goblet cells hyperplasia, proinflammatory cytokines, and nuclear factor kappa B (NFκB) p65 immunocontent in the lung. Moreover, 3-MA was able to improve oxidative stress, mitochondrial energy metabolism, and Na

Topics: Adenine; Animals; Anti-Asthmatic Agents; Asthma; Autophagy; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Energy Metabolism; Eosinophil Peroxidase; Eosinophils; Extracellular Traps; Female; Goblet Cells; Lung; Mice; Mice, Inbred BALB C; Mitochondria; Ovalbumin; Oxidative Stress; Reactive Oxygen Species; Transcription Factor RelA

A large proportion of asthmatic patients use complementary and alternative medicine (CAM) for the treatment of their disease. Various pharmacological effects, including anti-inflammatory properties, have been described for Portulaca oleracea (P. oleracea).. The study intended to evaluate the effects of an extract of P. oleracea on interleukin 4 (IL-4) and interferon-gamma (INF-γ) and on the INF-γ /IL4 ratio, as an index for the balance of T helper 1 and 2 (Th1/Th2) cells, in bronchoalveolar lavage fluid (BALF) as well as on tracheal responsiveness (TR) in a rat model of asthma.. The research team performed an animal study.. Forty-eight male Wistar rats, each weighing 220 ± 50 g, were included in the study.. The rats were randomly divided into 6 groups: (1) a control group that was not induced with asthma and that received no treatments (C group), (2) a group induced with asthma that received no treatments (A group), (3) an intervention group induced with asthma and treated with one mg/ml of P. oleracea extract (PO 1 group), (4) an intervention group induced with asthma and treated with 2 mg/ml of P. oleracea extract (PO 2 group), (5) an intervention group induced with asthma and treated with 4 mg/ml of P. oleracea extract (PO 4 group), and (6) a positive control group induced with asthma and treated with 1.25 μg/ml dexamethasone (D group). The asthma was induced using ovalbumin (OVA).. Tracheal responsiveness and the BALF levels of IL-4 and INF-γ and the INF-γ/IL4 ratio were measured.. Tracheal responsiveness to methacholine in the A and PO 1 groups and to OVA in the A, PO 1, and PO 2 groups as well as the BALF levels of IL-4 in the A, D, PO 1, PO 2, and PO 4 groups were significantly higher than that of the C group. The BALF level of INF-γ and the INF-γ/IL4 ratio were significantly lower in the A, D, PO 1, PO 2, and PO 4 groups than in the C group. Only treatment with the 2 higher concentrations of the extract caused a concentration-dependent increase in the INF-γ/IL4 ratio (P < .001). The effects of the 2 higher concentrations of the extract on INF-γ and IL-4 and on the INF-γ/IL4 ratio were significantly greater than those of the dexamethasone treatment (P < .001).. These results showed an immunomodulatory effect for the extract of P. oleracea with regard to an increased INF-γ/IL4 ratio, as an index of Th1/Th2, as well as a preventive effect on tracheal responsiveness, which was more specific than the effects of dexamethasone on the Th1/Th2 balance.

Topics: Animals; Asthma; Disease Models, Animal; Humans; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Portulaca; Rats; Rats, Wistar; Th1 Cells

Eriobotrya japonica leaves has a very long history of medicinal use as an anti-inflammatory and antitussive agent for bronchial inflammation.. The aim of this study was to evaluate the anti-inflammatory activities of Eriobotrya japonica (EJ) leaf water extract in an ovalbumin (OVA)-induced murine asthma model and human tracheal smooth muscle cell (HTSMC).. Mice were sensitized by intra peritoneal OVA and challenged with nebulized OVA. EJ extract was administered orally at various dose. Bronchoalveolar lavage fluid (BALF) was quantified for nitric oxide (NO), eosinophil peroxidase (EPO), interleukin (IL)-4, IL-13 level and immunoglobulin (Ig) E was quantified in serum. Lung tissue sections were stained with hematoxylin and eosin for assessment of inflammatory cell infiltration whereas mucus production and goblet cell hyperplasia were studied by periodic acid schiff staining. Western blot was done for analysis of pERK1/2 expression and NFκB translocation in HTSMC whereas iNOS and COX-2 expression in RAW264.7 cell.. EJ significantly reduced the levels of BALF's NO, EPO, MMPs, IL-4, IL-13, and serum IgE. It also decreases inflammatory cell infiltration and mucus production. EJ also attenuated the proliferation of HTSMC, inhibits overexpression of ERK 1/2 and translocation of NFκB in HTSMC as well as iNOS and COX-2 expression in RAW 264.7 cell.. Present study suggest that, EJ effectively protects against allergic airway inflammation thus possessing potential therapeutic option for allergic asthma management.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Eriobotrya; Female; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Plant Leaves; RAW 264.7 Cells

To investigate the effect of hydro-ethanolic extract of Zataria multiflora (Z. multiflora) on endothelin level, total and differential white blood cells (WBC) count of sensitized guinea pigs.. Five groups of guinea pigs sensitized to ovalbumin (OA) were given drinking water alone (group S), drinking water containing three concentrations of Z. multiflora (0.2, 0.4 and 0.8 mg/mL as groups S+Z1, S+Z2 and S+Z3) and dexamethasone (group S+D), n=6 for each group. The endothelin levels as well as total and differential WBC count in blood of sensitized and control guinea pigs were evaluated using enzyme linked immunosorbent assay method, and hemocytometer and Wright-Giemsa's staining of blood sample smear respectively.. Blood endothelin levels, total and most differential WBC count were increased but lymphocytes decreased in sensitized animals compared to controls (allP <0.01). In groups S+D, S+Z2 and S+Z3 endothelin level, total and differential WBC counts were significantly improved compared with group S (P <0.01). Although, all measured parameters in group S+Z1 was lower than group S+D (P <0.01), some parameters in group S+Z3 were greater than in group S+D (P <0.05 toP <0.01).. The results showed an anti-inflammatory effect of Z. multiflora extract in sensitized guinea pigs, which may suggest a therapeutic potential for the plant on asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Endothelins; Guinea Pigs; Iran; Leukocyte Count; Ovalbumin; Plant Extracts

Epidemiological studies have confirmed atmospheric PM. Rats were treat with ovalbumin (OVA) to establish asthma models. Notch signaling pathway inhibitor (DAPT) was injected intraperitoneally. Asthmatic and healthy rats were exposed to different concentrations of PM. Hes1 expression levels in healthy and asthma pathway inhibition groups were lower than those in control groups. Compared with control group, rats exposed to PM. PM

Topics: Animals; Asthma; Cholesterol, HDL; Cholesterol, LDL; Diamines; Disease Models, Animal; Dyslipidemias; Female; Gene Expression; Lipid Metabolism; Male; Ovalbumin; Particulate Matter; Rats; Rats, Wistar; Signal Transduction; Thiazoles; Transcription Factor HES-1; Triglycerides

Inflammatory airway diseases, such as asthma, cystic fibrosis (CF), and chronic obstructive pulmonary disease (COPD), are characterized by mucus hypersecretion and airway plugging. In both CF and asthma, enhanced expression of the Ca2+-activated Cl- channel TMEM16A is detected in mucus-producing club/goblet cells and airway smooth muscle. TMEM16A contributes to mucus hypersecretion and bronchoconstriction, which are both inhibited by blockers of TMEM16A, such as niflumic acid. Here we demonstrate that the FDA-approved drug niclosamide, a potent inhibitor of TMEM16A identified by high-throughput screening, is an inhibitor of both TMEM16A and TMEM16F. In asthmatic mice, niclosamide reduced mucus production and secretion, as well as bronchoconstriction, and showed additional antiinflammatory effects. Using transgenic asthmatic mice, we found evidence that TMEM16A and TMEM16F are required for normal mucus production/secretion, which may be due to their effects on intracellular Ca2+ signaling. TMEM16A and TMEM16F support exocytic release of mucus and inflammatory mediators, both of which are blocked by niclosamide. Thus, inhibition of mucus and cytokine release, bronchorelaxation, and reported antibacterial effects make niclosamide a potentially suitable drug for the treatment of inflammatory airway diseases, such as CF, asthma, and COPD.

Topics: Animals; Anoctamins; Anti-Inflammatory Agents; Asthma; Bronchi; Cell Line, Tumor; Cystic Fibrosis; Disease Models, Animal; Drug Repositioning; Goblet Cells; HEK293 Cells; Humans; Mice; Mice, Knockout; Mice, Transgenic; Mucus; Niclosamide; Ovalbumin; Pulmonary Disease, Chronic Obstructive; Signal Transduction

Airway inflammation and oxidative stress are the two major characteristics of asthma pathogenesis. Therefore, this study evaluated the protective effects of ascorbic acid in combination with calcitriol on the oxidative damages and inflammation in asthma model. All animals, except in the control group, were sensitized and challenged with ovalbumin. One day after the last challenge, samples of bronchoalveolar lavage fluid was collected for the assessment of total white blood cell counts and differential count of white blood cell and plasma was used for the measurement of pro-oxidant/antioxidant balance level. Lung tissue samples were also stored for examining peribronchial inflammatory cell infiltration, phosphorylated nuclear factor-kappa B expression and measurement of malondialdehyde level. Induction of asthma caused significant increases in total white blood cell counts, percentage of neutrophils and eosinophils and a decrease in the percentage of lymphocytes. Moreover, asthma resulted in significant increases of peribronchial inflammatory cell infiltration, phosphorylated nuclear factor-kappa B expression and malondialdehyde level. However, no significant changes were observed in pro-oxidant/antioxidant balance level with the induction of asthma. Co-administration of low doses of ascorbic acid and calcitriol returned all to the levels measured before sensitization and challenge. Combination of low doses of ascorbic acid with calcitriol improves mouse asthma model by a possible additive effects through the decrease of oxidative stress and inflammation.

Topics: Animals; Anti-Inflammatory Agents; Ascorbic Acid; Asthma; Bronchoalveolar Lavage Fluid; Calcitriol; Calcium Channel Agonists; Chronic Disease; Drug Therapy, Combination; Leukocyte Count; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Vitamins

Asthma is a complex chronic disease and the pathogenesis is still not entirely clear. In this study, we aimed to clarify the role and mechanism of miR-29b in the development of asthma. We observed that miR-29b levels were decreased in the lung and spleen of OVA-induced asthmatic mice. Reverse transcription-quantitative polymerase chain reaction and flow cytometry demonstrated that the inducible co-stimulator (ICOS) expression at mRNA and protein levels was elevated in the lung of asthmatic mice, and miR-29b expression in the lung of asthmatic mice was negatively associated with ICOS mRNA levels by Pearson Correlation analysis. Additional, flow cytometry showed that the percentage of CD4+ICOS+ T cells in the lung and spleen was regulated by miR-29b, and dual luciferase reporter assay confirmed ICOS was a target gene of miR-29b. Furthermore, miR-29b overexpression in asthmatic mice was induced with miR-29b agomir by intranasal administration; miR-29b alleviated total inflammatory cell infiltration and CCL24 levels, decreased IL-5 levels in bronchoalveolar lavage fluid and serum, and upregulated IFN-γ expression in serum. This study demonstrates that miR-29b targets ICOS, thereby reverses the imbalance of T helper 1 cells (Th1)/Th2 responses and decreases eosinophils recruitment in the airway, which are key features of allergic airway inflammation. Therefore, miR-29b might be an attractive candidate target for asthma treatment.

Topics: Allergens; Animals; Asthma; Cell Movement; Disease Models, Animal; Eosinophils; Female; Gene Expression Regulation; Humans; Inducible T-Cell Co-Stimulator Protein; Lung; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Respiratory Hypersensitivity; RNA, Small Interfering; Th1 Cells; Th1-Th2 Balance; Th2 Cells

Inflammation, reversible obstruction, and hyperresponsiveness of the airways are characteristic findings of bronchial asthma. Several evidence has demonstrated the involvement of matrix metalloproteinase-2 in allergic airway inflammation. Matrix metalloproteinase-2 may promote aberrant tissue remodeling in late stages of allergic airway inflammation. However, whether matrix metalloproteinase-2 is detrimental or protective in early stages of allergic airway inflammation remains unclear. To evaluate this here we compared the severity of allergic bronchial asthma between mice overexpressing human matrix metalloproteinase-2 and wild type mice. After sensitization and challenge with an allergen, mice overexpressing the human matrix metalloproteinase-2 showed a significant reduction in airway hyperresponsiveness and in the expression of Th2 cytokines and IgE compared to their wild type counterparts. An inhibitor of matrix metalloproteinases abolished this beneficial effect of human matrix metalloproteinase-2 overexpression. Allergen-sensitized and challenged human matrix metalloproteinase-2 transgenic mice had enhanced percentage of M1 macrophages with increased expression of inducible nitric oxide synthase and STAT1 activation in the lungs compared to their wild type counterparts. There was no difference in the percentage of regulatory T cells between mouse groups. The results of this study showed that matrix metalloproteinase-2 is protective in allergic bronchial asthma by promoting polarization of macrophages to M1 phenotype.

Topics: Allergens; Animals; Asthma; Cytokines; Disease Models, Animal; Female; Humans; Immunoglobulin E; Inflammation; Lung; Macrophages; Male; Matrix Metalloproteinase 2; Mice; Mice, Inbred C57BL; Mice, Transgenic; Middle Aged; Ovalbumin; Respiratory Hypersensitivity; T-Lymphocytes, Regulatory; Th2 Cells

In 2013, WHO estimated that approximately 235 million people suffered from asthma worldwide. Asthma is a hyper responsive disorder, which is related to an imbalance between the T‑helper type 1 and 2 cells (henceforth, Th1 and Th2, respectively). Allium hookeri is a plant that is widely used for culinary purposes and also in traditional Asian medicine. The present study was conducted to elucidate the anti‑asthmatic effects and mechanism of action of A. hookeri root extracts (AHRE) in an ovalbumin (OVA)‑induced asthma mouse model. The mice were divided into five groups, namely, the control, the OVA‑treated group, the dexamethasone‑treated group, the 30 mg/kg AHRE‑treated group, and the 300 mg/kg AHRE‑treated group. The total WBC count and the differential cell count in the bronchoalveolar fluid, the level of serum IgE, the histopathological changes in the lung, and changes in the cell surface molecules, the asthma‑related cytokine levels, and Th cell transcription factors were evaluated. AHRE significantly ameliorated asthmatic changes, such as the total WBC count, eosinophil count, and the level of IgE; in addition, it reduced mucus hypersecretion, epithelial hyperplasia, and eosinophil infiltration in the lungs. AHRE significantly inhibited the expression of CD68+ cells and MHC class II+ molecules, Th1 cell transcription factor (T‑bet) activation, Th2 cell transcription factor (GATA‑3) activation, and TNF‑α in the lung tissue. Furthermore, it suppressed cell surface molecules, such as CD4+and CD8+; Th1‑related cytokines, such as IFN‑γ and IL‑12p40; Th2‑related cytokines, such as IL‑4 and IL‑5; and Th17‑related cytokines, such as IL‑6 and TNF‑α, in a dose‑dependent manner. Thus, AHRE may be considered a promising anti‑asthmatic drug.

Topics: Allium; Animals; Asthma; Cytokines; Disease Models, Animal; Female; Immunomodulation; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Plant Roots; Th1 Cells; Th2 Cells

Our previous study showed that sulfatide-activated type II natural killer T (NKT) cells can prevent allergic airway inflammation in an ovalbumin (OVA)-induced murine model of asthma, but the underlying mechanism is unclear. Recently, sulfatide-activated type II NKT cells were shown to modulate the function of dendritic cells in experimental autoimmune encephalomyelitis and nonobese diabetic mice. Thus, it was hypothesized that sulfatide-activated type II NKT cells may modulate the function of lung dendritic cells (LDCs) in asthmatic mice. Our data showed that, in our mouse models, activation of type II NKT cells by sulfatide administration and adoptive transfer of sulfatide-activated type II NKT cells resulted in reduced expression of surface maturation markers and proinflammatory cytokine production of LDCs. LDCs from sulfatide-treated asthmatic mice, in contrast to LDCs from PBS-treated asthmatic mice, significantly reduced allergic airway inflammation in vivo. However, we found no influence of sulfatide-activated type II NKT cells on the phenotypic and functional maturation of bone marrow-derived dendritic cells in vitro. In addition, adoptive transfer of sulfatide-activated type II NKT cells did not influence the phenotypic and functional maturation of LDCs in CD1d

Topics: Animals; Asthma; Dendritic Cells; Female; Inflammation Mediators; Lung; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Natural Killer T-Cells; Ovalbumin; Sulfoglycosphingolipids

Accumulating evidence suggests that non-typeable Haemophilus influenzae (NTHi) infection drives the development of steroid-resistant allergic airway disease (SRAAD), exacerbates clinical symptoms, worsens quality of life, and accounts for most of the related healthcare burden. The poor understanding of the pathogenesis of SRAAD deters the development of more effective therapeutic strategies. Here, we established a murine model of NTHi infection-induced exacerbation of allergic airway disease. We showed that NTHi infection drove Th 17-mediated pulmonary neutrophilic inflammation, aggravated airway hyper-responsiveness, and upset the balance of MUC5AC and MUC5B expression. Dexamethasone treatment effectively inhibited the features of allergic airway disease but failed to reduce NTHi-induced exacerbation, which was associated with the hyper-phosphorylation of p38 mitogen-activated protein kinase (MAPK). Interestingly, inhibition of p38 using a specific inhibitor (SB203580) only partly suppressed the airway hyper-responsiveness and mucus hyper-secretion but failed to abrogate the infection-induced neutrophilic inflammatory response in SRAAD. However, SB203580 and dexamethasone co-treatment substantially suppressed all the features of NTHi-induced SRAAD. Our findings highlight the importance of p38 MAPK in the pathogenesis of NTHi-induced steroid resistance, and this combined treatment approach may be a novel strategy against steroid-resistant asthma.

Topics: Animals; Asthma; Dexamethasone; Disease Models, Animal; Drug Therapy, Combination; Female; Haemophilus influenzae; Humans; Imidazoles; Inflammation; Lung; Mice; Mucin 5AC; Mucin-5B; Neutrophils; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Pyridines; Respiratory Mucosa; Signal Transduction; Symptom Flare Up; Th17 Cells

Recent studies have reported recruitment of inflammatory monocytes by cytokines including chemokine (C-C motif) ligand 2 (CCL2) are critical in allergic responses. We aimed to investigate the role of inflammatory monocytes and CCL2 in mouse model with ovalbumin (OVA)-induced allergic asthma. Mice were sensitized with OVA to induce allergic asthma. The proportion of inflammatory cells in bronchoalveolar lavage fluid (BALF) and peritoneal lavage fluid (PLF) were measured by flow cytometry. The expression of CCL2 and CCL2 receptor (CCR2) were determined by qPCR and western blot. The concentrations of Type 1 helper T (Th1) and Type 2 helper T (Th2) cytokines in PLF were detected by ELISA. Inflammatory monocytes are recruited in PLF, and expression of CCL2 and CCR2 were elevated in OVA-induced mice. In addition, transfer of CCR2 knockdown inflammatory monocytes decreased the levels of allergic asthma biomarkers. Injection of anti-CCL2 or anti-CCR2 antibody decreased the proportion of eosinophils and inflammatory monocytes in BALF. Blockade of CCL2/CCR2 signaling pathway suppressed the allergen-induced Th2 cytokines and enhanced the levels of Th1-associated cytokines. Blockade of CCL2/CCR2 signaling pathway in sensitization-recruited inflammatory monocytes exhibits protective effects in mouse model of OVA-induced allergic asthma by inhibiting the Th2 inflammatory responses.

Topics: Animals; Antibodies; Asthma; Cell Movement; Chemokine CCL2; Mice; Monocytes; Ovalbumin; Receptors, CCR2; Signal Transduction; Th1 Cells; Th2 Cells

Asthma is a common chronic airway inflammatory disease, characterized by airway inflammation and remodeling. Piperlongumine (PL) has a number of physiological and pharmacological properties. However, the anti‑asthmatic effect of PL has not been reported to date. In the present study, ovalbumin (OVA) was used to sensitize and challenge mice to induce asthma. The results revealed that PL pretreatment reduced OVA‑induced airway inflammatory cell infiltration, reduced Th2 cytokine expression, both in the bronchoalveolar lavage fluid and in lung tissues, reduced the serum IgE level, pro‑inflammatory cytokine [tumor necrosis factor (TNF)‑α and interleukin (IL)‑6] and intercellular adhesion molecule expression, as well as nuclear factor (NF)‑κB activation. In addition, PL also mitigated OVA‑induced goblet cell metaplasia, inhibited mucus protein secretion, mitigated airway fibrosis and downregulated fibrosis marker expression. It was also demonstrated that PL inhibited TNF‑α induced inflammatory cytokine expression and NF‑κB activation in vitro. Taken together, the findings of the present study indicated that PL can reduce OVA‑induced airway inflammation and remodeling in asthmatic mice, and that these effects may be mediated by inhibiting NF‑κB signaling.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Line; Cytokines; Dioxolanes; Fibrosis; Humans; Inflammation; Interleukin-6; Lung; Male; Mice; Mice, Inbred C57BL; Mucus; NF-kappa B; Ovalbumin; Signal Transduction; Th2 Cells

We investigated the antiasthmatic effect of mogroside V (Mog V) in mice with ovalbumin (OVA)-induced asthma. Administration of Mog V effectively attenuated OVA-induced airway hyperresponsiveness and reduced the number of inflammatory cells in bronchoalveolar lavage fluid (BALF). Histological examination showed that Mog V reduced the inflammatory infiltration of the lungs in the asthmatic mice. ELISAs suggested that Mog V effectively decreased the levels of IL-4, IL-5, and IL-13 in BALF and serum levels of OVA-specific IgE and IgG1 in the asthmatic mice. A quantitative reverse-transcription PCR assay also indicated that Mog V decreased the mRNA levels of IL-17A, IL-23, and RORγt in the lungs of the asthmatic mice (the opposite effect on Foxp3 mRNA). Furthermore, Mog V significantly reduced the OVA-induced activation of NF-κB in the lungs. This study indicates that Mog V alleviates OVA-induced inflammation in airways, and this effect is associated with a reduction in NF-κB activation. PRACTICAL APPLICATIONS: A traditional Chinese medicine herb has been reported to have a strong curative effect on asthma in clinical practice. Siraitia grosvenorii is known in China as a functional food product with the ability to improve lung function. Mogroside V is a triterpene glycoside isolated from S. grosvenorii. Nonetheless, the antiasthmatic effect of mogroside V has not been evaluated yet. The aim of this study was to investigate the antiasthmatic activity of mogroside V in mice with chemically induced asthma. The data from this study will provide some scientific evidence supporting wider use of S. grosvenorii in functional foods.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Eosinophils; Gene Expression Regulation; Lymphocytes; Macrophages; Male; Mice; Molecular Structure; Neutrophils; Ovalbumin; Random Allocation; Triterpenes

Butylphthalide (NBP) is a phthalide compound contained in Angelicae Sinensis Radix which is one of the most widely used traditional Chinese medicines. This study aims to explore the therapeutic effect of NBP on airway inflammation, mucus hypersecretion and their possible mechanism in asthma mice. BALB/c mice were sensitized and challenged with ovalbumin (OVA) for establishment of asthma model and then treated with NBP during day 22-77. The pulmonary function of the mice was determined, and the pathology of lung tissue and goblet cell hyperplasia were observed through analyzing inflammation scores and goblet cell percentage, respectively. Cytokine IL-4, IL-8, IL-13 and tumor necrosis factor-alpha (TNF-α) in bronchoalveolar lavage fluid (BALF) and total immunogloblin E (T-IgE) and OVA-specific IgE in serum were examined by enzyme-linked immunosorbent assay (ELISA). The expressions of Mucin 5AC (Muc5ac) and nuclear transcription factor-kappa B (NF-κB) in lung tissues were evaluated by immunohistochemistry, western blot and real-time polymerase chain reaction (RT-PCR). The results show that 50 mg/kg NBP significantly reduced OVA-induced increase in inflammation scoring, goblet cell percentage and mucus secretion of airway tissue, and improved the pulmonary function. NBP could also decrease IL-4, IL-8 IL-13, and TNF-α in BALF and T-IgE and OVA-specific IgE in serum. The expression of Muc5ac and NF-κB in lung tissue was significantly down-regulated after NBP treatment. This study suggested that NBP may effectively inhibit airway inflammation and mucus hypersecretion in asthma by modulating NF-κB activation.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Benzofurans; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Leukocyte Count; Lung; Mice, Inbred BALB C; Mucin 5AC; Mucus; NF-kappa B; Ovalbumin

Helminths and their products can shape immune responses by modulating immune cells, which are dysfunctional in inflammatory diseases such as asthma. We previously identified SJMHE1, a small molecule peptide from the HSP60 protein of Schistosoma japonicum. SJMHE1 can inhibit delayed-type hypersensitivity and collagen-induced arthritis in mice. In the present study, we evaluated this peptide's potential intervention effect and mechanism on ovalbumin-induced asthma in mice. SJMHE1 treatment suppressed airway inflammation in allergic mice, decreased the infiltrating inflammatory cells in the lungs and bronchoalveolar lavage fluid, modulated the production of pro-inflammatory and anti-inflammatory cytokines in the splenocytes and lungs of allergic mice, reduced the percentage of Th2 cells and increased the proportion of Th1 and regulatory T cells (Tregs). At the same time, Foxp3 and T-bet expression increased, and GATA3 and RORγt decreased in the lungs of allergic mice. We proved that SJMHE1 can interrupt the development of asthma by diminishing airway inflammation in mice. The down-regulation of Th2 response and the up-regulation of Th1 and Tregs response may contribute to the protection induced by SJMHE1 in allergic mice. SJMHE1 can serve as a novel therapy for asthma and other allergic or inflammatory diseases.

Topics: Animals; Asthma; Cytokines; Forkhead Transcription Factors; GATA3 Transcription Factor; Gene Expression Regulation; Hypersensitivity; Inflammation; Lung; Lymphocyte Subsets; Mice; Mice, Inbred BALB C; Nuclear Receptor Subfamily 1, Group F, Member 3; Ovalbumin; Peptides; RNA, Messenger; Schistosoma japonicum; Spleen; T-Box Domain Proteins

Although the bulk of research into the biology of serotonin 5-HT. An 18-week ovalbumin challenge period was performed to generate persistent, chronic asthma in BALB/c mice. Four once daily intranasal treatments of (R)-DOI were administered one week after allergen cessation, with respiratory parameters being measured by whole-body plethysmography (WBP). Cytokine and chemokine levels were measured by quantitative real-time polymerase chain reaction (qRT-PCR) in homogenized lung tissue, bronchoalveolar (BALF) fluid was analyzed for chemokine modulation by multiplex assays, and Periodic Acid-Schiff and Masson's Trichrome staining was performed to determine goblet cell infiltration and overall changes to lung morphology.. 5-HT. Overall, these data provide support for the therapeutic potential of (R)-DOI and 5-HT

Topics: Airway Remodeling; Amphetamines; Animals; Asthma; Chronic Disease; Female; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; Pneumonia; Receptors, Serotonin, 5-HT2; Serotonin 5-HT2 Receptor Agonists

This study focused on investigating whether NS8593 reverses airway smooth muscle (ASM) contraction and the underlying mechanism.. ASM contraction in mouse tracheal rings and lung slices was measured. Currents mediated by voltage dependent Ca. High K. Our results indicate that NS8593 inhibits LVDCCs and NSCCs, resulting in decreases of intracellular Ca

Topics: 1-Naphthylamine; Animals; Anti-Allergic Agents; Asthma; Calcium; Calcium Channels, L-Type; Male; Mice; Mice, Inbred BALB C; Muscle Contraction; Muscle Relaxation; Muscle, Smooth; Ovalbumin

Pistacia weinmannifolia (Anacardiaceae) has been used in herbal medicine for the treatment of influenza, dysentery and enteritis in China. It was recently observed that P. weinmannifolia root extract (PWRE) exerts anti‑inflammatory effects both in in vitro and in vivo models. Based on the results from previous studies, the present study investigated the protective effect of PWRE on airway inflammation and mucus hypersecretion. Treatment with PWRE significantly decreased the number of eosinophils and the levels of Th2 cytokines, such as interleukin (IL)‑4, IL‑5 and IL‑13, in the bronchoalveolar lavage fluid (BALF) of OVA‑exposed mice. PWRE decreased the high serum levels of total and OVA‑specific immunoglobulin E. PWRE also effectively inhibited the influx of inflammatory cells into the lung, as well as airway mucus hypersecretion. In addition, the increased level of monocyte chemoattractant protein‑1 was significantly decreased with the PWRE treatment in the BALF of OVA‑exposed mice and in lipopolysaccharide‑stimulated RAW264.7 macrophages. These protective effects of PWRE on OVA‑induced pulmonary inflammation were accompanied by the downregulation of mitogen associated protein kinases and nuclear factor‑κB activation. Thus, the results from the present study indicate that PWRE could be valuable adjuvant for the treatment of asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Disease Models, Animal; Gene Expression; Humans; Lung; Mice; NF-kappa B; Ovalbumin; Pistacia; Plant Extracts; Plant Roots; Pneumonia; RAW 264.7 Cells

Saxagliptin (Saxa), a dipeptidyl dipeptidase-4 (DPP-4) inhibitor, is widely used for the treatment of type 2 diabetes mellitus. It has been documented to have immunomodulatory and anti-inflammatory actions. Our objective was to delineate the protective effect and the underlying mechanism of Saxa-in comparison with Dexamethasone (Dexa) - in airway inflammation induced by ovalbumin (OVA) in mice.. Mice were OVA-sensitized and challenged for the induction of acute asthma. Mice were orally administrated Saxa or Dexa. Total and differential cell counts, lactate dehydrogenase (LDH) and total protein concentrations were assessed in bronchoalveolar lavage fluid (BALF). The toll-like receptor 4 (TLR4), nuclear factor-kappa B (NF-kB), reduced glutathione (GSH), and total nitrate/nitrite products (NOx) levels as well as myeloperoxidase (MPO) activity in lung tissues were measured. Histopathological examination of the lung specimens was carried out using the hematoxylin and eosin (H & E) staining.. Histopathological examination revealed that both Saxa and Dexa ameliorated OVA-induced inflammatory changes and significantly reduced total and differential leukocyte counts, LDH and total protein level in BALF upon comparison with OVA group. In addition, both treatments significantly mitigated OVA-induced oxidative stress as evidenced by diminished lung NOx level and MPO activity and elevated GSH level. The elevation of TLR4 and NF-kB levels in lung tissue were ameliorated by Saxa and Dexa administration.. Saxa had marked antiasthmatic effect in OVA-induced allergic asthma through modulation of TLR4 and NF-κB signaling. Also, Saxa may represent a promising therapeutic agent for acute allergic asthma.

Topics: Acute Disease; Adamantane; Animals; Asthma; Bronchoalveolar Lavage Fluid; Dexamethasone; Dipeptides; Dipeptidyl-Peptidase IV Inhibitors; L-Lactate Dehydrogenase; Lung; Male; Mice; NF-kappa B; Nitric Oxide; Ovalbumin; Peroxidase; Toll-Like Receptor 4

T helper 2 (Th2) lymphocytes and associated interleukin (IL) 4 and IL-13 play crucial roles in asthma pathogenesis. In this study, we explored an adeno-associated virus 5 (AAV5) based gene therapy by delivering truncated IL-4 protein to antagonize IL-4 receptor α chain and interrupt asthmatic signal pathway.. A recombinant adeno-associated virus 5 (AAV5) vector harboring a truncated mouse IL-4 gene (AAV5-mIL-4ΔC22) was prepared. Western blotting showed that the IL-4 mutant protein lacking the C-terminal 22 amino acids was expressed well in AAV5-mIL-4ΔC22 infected 16HBE and BEAS-2B cells. AAV5-drivn green fluorescent protein (AAV5-GFP) served as a control. The biodistribution of vector DNA after AAV5 vector aerosol inhalation was examined by PCR and the result showed that foreign DNA was detectable in the lungs but not in other organs including gonads. The aerosol inhalation-mediated delivery of AAV5-expressed antagonistic IL-4 mutant protein improved the lung function of ovalbumin-induced asthma mice.. The inhalation of aerosolized AAV5-mIL-4ΔC22 significantly improved the lung function and modulated the immune cell infiltration and associated cytokine expression in the bronchoalveolar lavage fluid (BALF) of ovalbumin-induced asthma mice.

Topics: Administration, Inhalation; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dependovirus; Disease Models, Animal; Female; Genetic Therapy; Interleukin-4; Interleukin-4 Receptor alpha Subunit; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Parvovirinae; Tissue Distribution

Asthma has affected more than 300 million people worldwide and is considered one of the most debilitating global public health problems based on a recent statistical report from the Global Initiative for Asthma. Inflammation of the airways leads to the various interrelated mechanisms of innate and adaptive immunity acting mutually with the epithelium of the respiratory organ. Fucoxanthin is an orange or brown pigment which is naturally found in various seaweeds. To the best of our knowledge, there are no scientific claims or evidence of the curative effects of fucoxanthin against asthma. Hence, this present research was designed to investigate the curative activity of fucoxanthin against ovalbumin-induced asthma in a mouse model. Fucoxanthin (50 mg/kg) showed significant (P < 0.001) antiasthma activity. It effectively decreased intracellular secretion of reactive oxygen species and increased antioxidant enzyme activity. Fucoxanthin also decreased inflammatory cytokine markers in bronchoalveolar lavage fluid. Because fucoxanthin showed effective antiasthma activity against ovalbumin-induced asthma in experimental animals, further research on this natural antioxidant could lead to development of a novel drug for the treatment of asthma in humans.

Topics: Animals; Anti-Asthmatic Agents; Antioxidants; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Cytokines; Inflammation; Male; Mice; Ovalbumin; Reactive Oxygen Species; Xanthophylls

Allergic asthma is a chronic inflammatory disease and involves many cells and cellular components. Cataract is a condition that affects the transparency of the lens, which the opacity of the lens caused by any innate or acquired factor degrades its transparency or changes in color. During the establishment of asthma model of rats with chicken ovalbumin nebulization, it was found that asthmatic rats were more likely to have monocular or binocular cataract symptoms than normal rats. Considering that they are all induced by immune imbalance, inflammation, etc., there may be some correlation in the mechanism, and many clues showed that both diseases are associated with activation of the PI3K-AKT-mTOR signaling pathway. Therefore, we hypothesized that the PI3K-AKT-mTOR signaling pathway produces inflammatory or immune imbalance based on allergy leading to cataract.

Topics: Animals; Asthma; Cataract; Disease Models, Animal; Hypersensitivity; Inflammation; Male; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Rats, Wistar; TOR Serine-Threonine Kinases

Glucocorticoids are among the most effective drugs to treat asthma. However, the severe adverse effects associated generate the need for its therapeutic optimization. Conversely, though histamine is undoubtedly related to asthma development, there is a lack of efficacy of antihistamines in controlling its symptoms, which prevents their clinical application. We have reported that antihistamines potentiate glucocorticoids' responses in vitro and recent observations have indicated that the coadministration of an antihistamine and a synthetic glucocorticoid has synergistic effects on a murine model of allergic rhinitis. Here, the aim of this work is to establish if this therapeutic combination could be beneficial in a murine model of asthma. We used an allergen-induced model of asthma (employing ovalbumin) to evaluate the effects of the synthetic glucocorticoid dexamethasone combined with the antihistamine azelastine. Our results indicate that the cotreatment with azelastine and a suboptimal dose of dexamethasone can improve allergic lung inflammation as shown by a decrease in eosinophils in bronchoalveolar lavage, fewer peribronchial and perivascular infiltrates, and mucin-producing cells. In addition, serum levels of allergen-specific IgE and IgG1 were also reduced, as well as the expression of lung inflammatory-related genes IL-4, IL-5, Muc5AC, and Arginase I. The potentiation of dexamethasone effects by azelastine could allow to reduce the effective glucocorticoid dose needed to achieve a therapeutic effect. These findings provide first new insights into the potential benefits of glucocorticoids and antihistamines combination for the treatment of asthma and grants further research to evaluate this approach in other related inflammatory conditions.

Topics: Administration, Intranasal; Animals; Anti-Asthmatic Agents; Asthma; Dexamethasone; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Synergism; Drug Therapy, Combination; Female; Glucocorticoids; HEK293 Cells; Histamine H1 Antagonists, Non-Sedating; Humans; Lung; Mice; Ovalbumin; Phthalazines; Receptors, Glucocorticoid; Transcriptional Activation

Topics: Animals; Antioxidants; Asthma; Cell Line; Collagen; Drug Evaluation, Preclinical; Flavanones; Immunoglobulin E; Mice; Mucus; NF-kappa B; Nitric Oxide Synthase Type II; Ovalbumin; Phytotherapy; Plant Extracts; Scutellaria baicalensis; Signal Transduction

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; Hypersensitivity; Immunoglobulin E; Immunophenotyping; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Nickel; Ovalbumin; Particle Size

The tuberous roots of Liriope platyphylla (Liriopis Tuber; LT) is traditionally used in Korean Medicine for treating colds, cough, and sputum production. In this study, we investigated the effect of spicatoside A isolated from LT methanol extract on ovalbumin (OVA)-sensitized/challenged asthmatic mice. For induction of allergic asthma, BALB/c mice were sensitized with OVA by an intraperitoneal injection at three times a week, and then challenged into the nasal cavities using a nebulizer. Spicatoside A at dose of 1mg/kg body weight was treated in mice with an oral administration once daily for a week during OVA challenge. The concentrations of OVA-specific IgE, IL-4, IL-5 and IL-13 were measured in the sera or bronchoalveolar lavage fluids (BALF) of mice by enzyme-linked immunosorbent assay (ELISA). The numbers of total cells, macrophages, lymphocytes, neutrophils and eosinophils were counted in BALFs using Diff-Quik staining, and histopathological changes of lung tissues were observed by hematoxylin and eosin (H&E), Periodic acid Schiff (PAS) and Masson's trichrome staining. The purity of spicatoside A was 98.1% with a white powder (yield: 465.6mg). The treatment of spicatoside A in asthmatic mice significantly decreased the production of allergic mediator, OVA-specific IgE and Th2 cytokines, IL-4, IL-5 and IL-13 in sera and BALF. The numbers of inflammatory cells such as macrophages, lymphocytes, neutrophils and eosinophils in BALF of asthmatic mice were significantly reduced by the treatment of spicatoside A. Furthermore, the treatment of spicatoside A in asthmatic mice inhibited the structural damages of lung tissues with thickened bronchiolar epithelium and infiltration of inflammatory cells, the accumulation of mucus by the goblet cells hyperplasia and collagen in the bronchioles. These results suggest that spicatoside A of LT has a preventive effect on allergic asthma through the inhibition of lung inflammation and allergic response.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Immunoglobulin E; Inflammation; Liriope Plant; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Saponins

The respiratory system is exposed to various allergens via inhaled and intranasal routes. Murine models of allergic lung disease have been developed to clarify the mechanisms underlying inflammatory responses and evaluate the efficacy of novel therapeutics. However, there have been no comparative studies on differences in allergic phenotypes following inhaled vs. intranasal allergen challenge. In this study, we compared the asthmatic features of mice challenged via different routes following allergen sensitization and investigated the underlying mechanisms.. To establish ovalbumin (OVA)-induced allergic asthma models, BALB/c mice were sensitized to 20 μg OVA with 1 mg aluminum hydroxide by the intraperitoneal route and then challenged by inhalation or intranasal administration with 5% OVA for 3 consecutive days. Cellular changes and immunoglobulin (Ig) E levels in bronchoalveolar lavage fluid (BALF) and serum, respectively, were assessed. Histological changes in the lungs were examined by hematoxylin and eosin (H&E) and periodic acid Schiff (PAS) staining. Levels of T helper (Th)2 cytokines including interleukin (IL)-4, -5, and -13 in BALF and epithelial cytokines including IL-25 and -33 in BALF and lung tissues were measured by enzyme-linked immunosorbent assay and western blotting. Airway hyperresponsiveness (AHR) was evaluated by assessing airway resistance (Rrs) and elastance (E) via an invasive method.. OVA-sensitized and challenged mice showed typical asthma features such as airway inflammation, elevated IgE level, and AHR regardless of the challenge route. However, H&E staining showed that inflammation of pulmonary vessels, alveolar ducts, and alveoli were enhanced by inhaled as compared to intranasal OVA challenge. PAS staining showed that intranasal OVA challenge induced severe mucus production accompanied by inflammation in bronchial regions. In addition, Th2 cytokine levels in BALF and AHR in lung were increased to a greater extent by inhalation than by intranasal administration of OVA. Epithelial cytokine expression, especially IL-25, was increased in the lungs of mice in the inhaled OVA challenge group.. OVA-sensitized mice exhibit different pathophysiological patterns of asthma including expression of epithelial cell-derived cytokines depending on the OVA challenge route. Thus, some heterogeneous phenotypes of human asthma can be replicated by varying the mode of delivery after OVA sensitization.

Topics: Administration, Inhalation; Administration, Intranasal; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Interleukin-17; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phenotype; T-Lymphocytes

β. We established OVA-sensitized and -challenged acute and chronic asthma mice models to explore the expression of Epac at first. Then, airway inflammation and airway hyperresponsiveness in acute asthma mice model and airway remodeling in chronic asthma mice model were observed respectively after treatment with Epac-selective cAMP analogue 8-pCPT-2'-O-Me-cAMP (8pCPT) and Epac inhibitor ESI-09. Next, the effects of 8pCPT and ESI-09 on the proliferation and apoptosis of in vitro cultured mouse airway smooth muscle cells (ASMCs) were detected with CCK-8 assays and Annexin-V staining. Lastly, the effects of 8pCPT and ESI-09 on store-operated Ca. We found that in lung tissues of acute and chronic asthma mice models, both mRNA and protein expression of Epac1 and Epac2, two isoforms of Epac, were lower than that of control mice. In acute asthma mice model, the airway inflammatory cell infiltration, Th2 cytokines secretion and airway hyperresponsiveness were significantly attenuated by 8pCPT and aggravated by ESI-09. In chronic asthma mice model, 8pCPT decreased airway inflammatory cell infiltration and airway remodeling indexes such as collagen deposition and airway smooth muscle cell proliferation, while ESI-09 increased airway inflammation and airway remodeling. In vitro cultured mice ASMCs, 8pCPT dose-dependently inhibited, whereas ESI-09 promoted ASMCs proliferation. Interestingly, 8pCPT promoted the apoptosis of ASMCs, whereas ESI-09 had no effect on ASMCs apoptosis. Lastly, confocal Ca. Our results suggest that Epac has a protecting effect on asthmatic airway inflammation and airway remodeling, and Epac reduces ASMCs proliferation by inhibiting SOCE in part.

Topics: Airway Remodeling; Animals; Apoptosis; Asthma; Calcium Signaling; Cell Proliferation; Cells, Cultured; Cyclic AMP; Disease Models, Animal; Female; Guanine Nucleotide Exchange Factors; Hydrazones; Inflammation Mediators; Isoxazoles; Lung; Mice, Inbred BALB C; Myocytes, Smooth Muscle; Ovalbumin; Pneumonia; Respiratory Hypersensitivity

Schizophyllum commune is a ubiquitous basidiomycetous fungus typically found across the world, which has been detected in indoor and outdoor air. Some studies indicated that sensitization to S. commune is correlated with asthma severity in patients. Patients with chronic severe or acute fatal asthma have neutrophil-dominant airway inflammation. We hypothesized that S. commune can exacerbate asthma. To test this hypothesis, we evaluated the direct immunomodulatory activities of S. commune in allergic airway inflammation induced by non-fungal sensitization. Ovalbumin (OVA)-induced asthma model mice were generated using wild-type (WT) and Il-17a

Topics: Animals; Asthma; Disease Models, Animal; Female; Interleukin-17; Mice, Inbred C57BL; Neutrophils; Ovalbumin; Pneumonia; Schizophyllum; Th17 Cells

Anxiety is prevalent in asthma, and is associated with disease severity and poor quality of life. However, no study to date provides direct experimental evidence for the effect of allergic inflammation on the structure and function of medial prefrontal cortex (mPFC) and amygdala, which are essential regions for modulating anxiety and its behavioral expression. We assessed the impact of ovalbumin (OVA)-induced allergic inflammation on the appearance of anxiety-like behavior, mPFC and amygdala volumes using MRI, and the mPFC-amygdala circuit activity in sensitized rats. Our findings exhibited that the OVA challenge in sensitized rats induced anxiety-like behavior, and led to more activated microglia and astrocytes in the mPFC and amygdala. We also found a negative correlation between anxiety-like behavior and amygdala volume. Moreover, OVA challenge in sensitized rats was associated with increases in mPFC and amygdala activity, elevation of amygdala delta-gamma coupling, and the enhancement of functional connectivity within mPFC-amygdala circuit - accompanied by an inverted direction of information transferred from the amygdala to the mPFC. We indicated that disrupting the dynamic interactions of the mPFC-amygdala circuit may contribute to the induction of anxiety-related behaviors with asthma. These findings could provide new insight to clarify the underlying mechanisms of allergic inflammation-induced psychiatric disorders related to asthma.

Topics: Allergens; Amygdala; Animals; Anxiety; Asthma; Behavior, Animal; Disease Models, Animal; Inflammation; Lung; Magnetic Resonance Imaging; Male; Maze Learning; Ovalbumin; Prefrontal Cortex; Rats; Rats, Wistar

To establish and evaluate an ovalbumin (OVA)-induced bronchial asthma model in mice with intrauterine growth retardation (IUGR), and to explore the molecular mechanism of relationship between IUGR and asthma.. A total of 16 pregnant BALB/c female mice were divided into a low-protein diet group (n=8) and a normal-protein diet group (n=8), which were fed with low-protein (8%) diet and normal-protein (20%) diet respectively. The neonatal mice were weighed 6 hours after birth. Sixteen male neonatal mice with IUGR were randomly chosen from the low-protein diet group and enrolled in the IUGR group, and 16 male neonatal mice from the normal-protein diet group were enrolled in the control group. Blood samples were collected from the mice in both groups for testing of blood glucose. Enzyme-linked immunosorbent assay (ELISA) was used to determine serum insulin level. The mice in the control group were randomized into a control + PBS group and a control + OVA group (n=8 each). The mice in the IUGR group were randomized into an IUGR + PBS group and an IUGR + OVA group (n=8 each). Six-week-old mice in the control + OVA and IUGR + OVA groups were subjected to intraperitoneal injection of 2 mg/mL OVA for sensitization and aerosol inhalation of 1% OVA for challenge. Mice in the control + PBS group and the IUGR + PBS group were treated with an equivalent amount of PBS. ELISA was used to determine serum IgE level in the mice in each group. Bronchoalveolar lavage fluid (BLF) was collected from the mice in each group for cell counting. The lung tissue of the mice in each group was stained with hematoxylin and eosin to observe pathological changes.. The body weight at 6 hours after birth was significantly lower for neonatal mice in the low-protein diet group compared with those in the normal-protein diet group (P<0.01). The IUGR group had a significantly lower serum insulin level than the control group (P<0.01). The IUGR + PBS group had a significantly lower IgE level than the control + PBS group (P<0.01). Compared with the control + PBS and IUGR + PBS groups, the control + OVA and IUGR + OVA groups had a significantly increased IgE level, and the IgE level was significantly higher in the IUGR + OVA group than in the control + OVA group (P<0.01). Compared with the control + PBS and IUGR + PBS groups, the control + OVA and IUGR + OVA groups had significantly increased counts of leukocytes, eosinophils, lymphocytes, and macrophages in the BLF (P<0.01). The pulmonary alveoli of OVA-induced IUGR mice showed massive inflammatory cell infiltration and damage of intercellular continuity. Meanwhile, airway epithelial cell proliferation, bronchial wall thickening, bronchial lumen narrowing, and massive inflammatory cell infiltration around the bronchi and the vascular wall were observed.. An OVA-induced bronchial asthma model has been successfully established in the mice with IUGR induced by low-protein diet, which provides a basis for further study of the molecular mechanism of relationship between IUGR and airway inflammation.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Fetal Growth Retardation; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin

Allergic asthma, characterized by chronic airway Th2-dominated inflammation, is associated with an increased risk of infection; however, the underlying mechanisms are unclear. Forkhead box protein A2 (Foxa2) plays a critical role in Th2 inflammation and is associated with pulmonary defenses. To determining the role of Foxa2 in Th2-dominated lung inflammation against the invading bacteria, we established a mouse OVA-sensitized model, an Escherichia coli lung invasion model, and mice with conditional deletion of Foxa2 in respiratory epithelial cells. The number of bacteria in the lung tissue was counted to assess clearance ability of lung. Lung inflammation and histopathology was evaluated using HE and PAS staining. It was found that OVA-sensitized mice had decreased E. coli clearance, reduced Foxa2 expression, and decreased DEFB1 secretion. Conditional deletion of Foxa2 in respiratory epithelial cells led to decreased clearance of E. coli and impaired secretion of DEFB1, similar to the OVA-induced allergic condition. The impaired secretion of DEFB1 may be responsible for the increased risk of infection in the Th2-dominated airway inflammation. Dual luciferase assay demonstrated that Foxa2 regulates DEFB1 expression by affecting its promoter activity in HBE cells. Our study indicated that Foxa2 plays an important role in Th2-dominated airway inflammation against invading bacteria. Conditional deletion of Foxa2 in respiratory epithelial cells can reduce pulmonary's defense against bacterial invasion by inhibiting DEFB1expression.

Topics: Animals; Asthma; beta-Defensins; Cell Line; Disease Models, Animal; Escherichia coli Infections; Female; Gene Deletion; Gene Expression Regulation; Hepatocyte Nuclear Factor 3-beta; Humans; Mice; Ovalbumin; Th2 Cells

Statins provide greater protection than predicted from just cholesterol-lowering effects, which is possibly mediated by other pleiotropic actions. This study aimed to examine the possible interaction effect of asthma on lipid profiles and evaluate the effect of rosuvastatin treatment on asthma. The animals were assigned into (1) control, (2) asthmatic, (3) hyperlipidemic, (4) asthmatic-hyperlipidemic, (5) rosuvastatin (40 mg/kg/day intraperitoneally, for 3 weeks)-treated asthmatic, (6) rosuvastatin-treated hyperlipidemic and (7) rosuvastatin-treated asthmatic-hyperlipidemic groups. Tracheal responsiveness to methacholine and ovalbumin, total and differential WBC (white blood cell) counts, and oxidative stress markers in bronchoalveolar lavage fluid (BALF) were evaluated. In the asthmatic and asthmatic-hyperlipidemic groups, tracheal responsiveness to ovalbumin, total WBC count, numbers of eosinophils, neutrophils, and monocytes were higher than the control group (p<0.001). A left-ward shift in the concentration-response curves to methacholine, an increase in nitrite and malondialdehyde concentrations, and a decrease in total thiol content, superoxide dismutase and catalase activities were also observed in the asthmatic and asthmatic-hyperlipidemic groups compared to control group (p<0.01 to p<0.001). Beyond lipid-lowering effect in the treated hyperlipidemic and asthmatic-hyperlipidemic groups, rosuvastatin treatment decreased tracheal responsiveness to methacholine, reduced total WBC count, the numbers of eosinophils, neutrophils, and monocytes, as well as decreased malondialdehyde concentration, and increased total thiol content, superoxide dismutase and catalase activities in treated asthmatic and asthmatic-hyperlipidemic groups (p<0.05 to p<0.001). The improving effect of rosuvastatin on asthmatic and asthmatic-hyperlipidemic animals was shown due to pleiotropic mechanisms including the effect on airway hyperresponsiveness, lung inflammation, and oxidative stress.

Topics: Allergens; Animals; Anticholesteremic Agents; Asthma; Bronchoalveolar Lavage Fluid; Hyperlipidemias; Leukocyte Count; Lipids; Male; Methacholine Chloride; Ovalbumin; Oxidative Stress; Rats, Wistar; Rosuvastatin Calcium; Trachea

Allergic inflammation is a response of the body against pathogens by cytokine release and leucocyte recruitment. Recently, there was an increase in morbimortality associated with allergic inflammation, especially asthma. The treatment has many adverse effects, requiring the search for new therapies. Monoterpenes are natural products with anti-inflammatory activity demonstrated in several studies and can be an option to inflammation management. Thus, we investigated the effects of citronellol, α-terpineol and carvacrol on allergic inflammation. The model of asthma was established by OVA induction in male Swiss mice. The monoterpenes were administered (25, 50 or 100 mg/kg, i.p.) 1 h before induction. After 24hs, the animals were sacrificed to leucocytes and TNF-α quantification. Monoterpenes significantly decrease leucocyte migration and TNF-α levels, possibly by modulation of COX, PGE

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Asthma; Cyclooxygenase 2 Inhibitors; Cytokines; Dinoprostone; Disease Models, Animal; Inflammation; Male; Mice; Molecular Docking Simulation; Monoterpenes; Oils, Volatile; Ovalbumin; Receptors, Histamine H1; Spices; Tumor Necrosis Factor-alpha

Croton zehntneri Pax et Hoffm. is a Euphorbiaceae species, popularly known as "canela de cunhã," a native plant of northeastern Brazil, whose essential oil (EOCZ) shows relatively specific myorelaxant action for the smooth muscle of the airways and in the respiratory tract. Based on this information, EOCZ figures as a candidate for testing in the treatment of asthma, and the present study investigated the benefits of using EOCZ in an ovalbumin-induced asthma model.. 48 male BALB/c mice were divided into six groups (n = 8). In the ST, SO100, and SO300 groups, mice were sensitized and challenged with saline, and then treated with 200 µL of 0.1% Tween 80, 100 mg/kg EOCZ and 300 mg/kg EOCZ, respectively. In the OT, OO100, and OO300 groups, mice were sensitized and challenged with OVA, and then treated with 200 µL of 0.1% Tween 80, 100 mg/kg EOCZ and 300 mg/kg EOCZ, respectively.. Our results demonstrated significant changes in all respiratory mechanics variables analyzed between the OO300 and OT groups demonstrating the effectiveness of EOCZ to attenuate the OVA-induced lung injury. In addition, the use of EOCZ at a dose of 300 mg/kg showed an antioxidant effect and decreased inflammatory cells in the pulmonary parenchyma. In conclusion, our results demonstrated that EOCZ was able to improve the lesion in the respiratory system of mice subjected to OVA-induced asthma.. The antioxidant action of EOCZ was likely the main mechanism of action in the reversal of this lesion, so more tests should be performed for its confirmation.

Topics: Animals; Asthma; Brazil; Croton; Disease Models, Animal; Dose-Response Relationship, Drug; Humans; Lung; Lung Injury; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Oils, Volatile; Ovalbumin; Phytotherapy; Plant Leaves; Respiratory Mechanics

The effects of bone marrow-derived mesenchymal stem cells (BMSCs) and simvastatin combination therapy on serum total and specific IgE levels and lung IL-13 and TGF-β levels in sensitized mouse were examined.. Serum total and specific IgE levels as well as lung IL-13 and TGF-β levels were significantly increased in S group compared to control group (P < 0.001 for all cases). Treatment with BMSCs, simvastatin and their combination significantly decreased serum total and specific IgE levels (P < 0.05 to P < 0.01). However, IL-13 and TGF-β levels were significantly decreased by BMSCs and BMSC + simvastatin combination therapy (P < 0.05 for all cases). The effect of simvastatin and BMSCs combination therapy on serum specific IgE levels as well as lung IL-13 and TGF-β levels were significantly higher than the effect of BMSCs and simvastatin alone (P < 0.001 for IL-13 and P < 0.01 for other cases).. Simvastatin and BMSCs combination therapy affects serum IgE as well as lung IL-13 and TGFβ levels more than BMSC therapy and simvastatin therapy alone which may be due to increased BMSCs migration into the lung tissue.

Topics: Animals; Asthma; Bone Marrow; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Immunoglobulin E; Interleukin-13; Lung; Male; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Simvastatin; Transforming Growth Factor beta

Serum IL-22 levels are increased in patients with atopic dermatitis, which commonly precedes asthma in the atopic march. Epicutaneous sensitization in mice results in T. We sought to determine the role of IL-22 in antigen-driven airway allergic inflammation after inhalation challenge in epicutaneously sensitized mice.. Wild-type (WT) and Il22. Epicutaneous sensitization preferentially elicited an IL-22 response compared with intraperitoneal immunization. Intranasal challenge of mice epicutaneously sensitized with OVA elicited in the lungs Il22 mRNA expression, IL-22 production, and accumulation of CD3. Epicutaneous sensitization promotes generation of antigen-specific IL-22-producing T cells that promote airway inflammation and AHR after antigen challenge, suggesting that IL-22 plays an important role in the atopic march.

Topics: Allergens; Animals; Asthma; Dermatitis, Atopic; Disease Models, Animal; Humans; Hypersensitivity; Immunization; Inflammation; Interleukin-22; Interleukins; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Respiratory Hypersensitivity; Skin; Th2 Cells

To investigate the correlations among airway inflammation, airway epithelial injury and airway hyperresponsiveness (AHR) in asthmatic mice treated with dexamethasone.. Female BALB/c mice were sensitized with intraperitoneal and hypodermic injections of ovalbumin (OVA) and aluminum on days 0, 7 and 14, challenged with OVA starting on day 21 for 10 days, and treated with dexamethasone via intraperitoneal injection starting on day 28 for 3 days. Female C57BL/6 mice were treated intranasally with house dust mite (HDM) on days 1 and 14, challenged intranasally with HDM on days 21, 23, 25, 27 and 29, and treated with sivelestat (a selective neutrophil elastase inhibitor) via intraperitoneal injection after each challenge. Following the final challenge, enhanced pause (Penh) and differential cell counts in the broncho-alveolar lavage fluid were measured and the correlations were analyzed.. Compared with OVA-challenged BALB/c mice, the counterpart mice treated with dexamethasone showed reduced Penh and shedding of airway epithelial cells. In addition, we found that Penh 50 (an indicator of AHR) had positive correlations with airway neutrophils and shedding of airway epithelial cells, but no correlation with eosinophils, lymphocytes or macrophages. Moreover, shedding of airway epithelial cells had positive correlations with airway neutrophils, but no correlation with eosinophils, lymphocytes or macrophages. Further, sivelestat decreased Penh 50 and shed airway-epithelial cells in HDM-challenged C57BL/6 mice.. Collectively, our findings suggest that airway neutrophils and excessive shedding of airway epithelial cells, but not eosinophils, lymphocytes or macrophages, may be involved in AHR in asthmatic mice treated with dexamethasone.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Dexamethasone; Disease Models, Animal; Eosinophils; Female; Glycine; Inflammation; Lymphocytes; Macrophages; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Neutrophils; Ovalbumin; Pyroglyphidae; Respiratory Mucosa; Respiratory System; Sulfonamides

Mycoplasma pneumoniae, an atypical human pathogen, has been associated with asthma initiation and exacerbation. Asthmatic patients have been reported to have higher carriage rates of M pneumoniae compared with nonasthmatic subjects and are at greater risk for invasive respiratory infections.. We sought to study whether prior allergen sensitization affects the host response to chronic bacterial infection.. BALB/cJ and IL-4 receptor α. Anti-M pneumoniae antibody responses were decreased in allergen-sensitized, M pneumoniae-infected animals compared with control animals, but OVA-specific IgG responses were unaffected. Similar decreases in anti-S pneumoniae antibody levels were found in OVA-sensitized animals. However, M pneumoniae, but not S pneumoniae, infection augmented anti-OVA IgE antibody responses. Loss of IL-4 receptor signaling partially restored anti-M pneumoniae antibody responses in IgG. An established type 2-biased host immune response impairs the host immune response to respiratory bacterial infection in a largely pathogen-independent manner. Some pathogens, such as M pneumoniae, can augment ongoing allergic responses and inhibit pulmonary type 2 cytokine responses and allergic airway hyperreactivity.

Topics: Allergens; Animals; Asthma; Cytokines; Immunoglobulin G; Lung; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Pneumococcal Infections; Pneumonia, Mycoplasma; Receptors, Cell Surface; Respiratory Tract Infections

Reactive oxygen species (ROS) and epithelial-mesenchymal transition (EMT) play a critical role in transforming growth factor (TGF)-β1-mediated fibrotic airway remodeling in asthma. Polydatin (PD) is a small natural molecule in Chinese medicine; it is isolated from Polygonum cuspidatum and has antioxidative properties. In this study, we aimed to determine whether PD was protective against ROS-induced pulmonary fibrosis in asthma. Ovalbumin (OVA) was used to induce asthma in a mouse model that was treated with or without PD. We also created nuclear factor erythroid 2-related factor 2 (Nrf2) knockdown BEAS-2B cells and investigated whether PD reversed TGF-β1-induced pulmonary epithelial cell EMT by promotion of Nrf2-mediated antioxidation. Immunofluorescence showed that ROS and TGF-β1 expression was significantly increased in lung tissue from the OVA-induced asthma model. PD treatment inhibited activity of ROS and TGF-β1. Immunohistochemistry showed that PD treatment decreased OVA-induced lung ROS, TGF-β1 expression and fibroblasts. Western blotting showed that PD treatment reversed OVA-induced NADPH oxidase (NOX)1/4 expression by promoting Nrf2-mediated heme oxygenase-1 and NADPH dehydrogenase (quinone)-1 expression. PD treatment suppressed OVA-induced EMT and lung fibroblast protein expression in lung tissue. Nrf2 downregulation suppressed the protective effect of PD by promoting TGF-β1-induced ROS and EMT and accumulation of extracellular-matrix-related protein. All these data indicate that PD has potential therapeutic effects in asthma by promoting Nrf2-mediated antioxidation.

Topics: Airway Remodeling; Animals; Antioxidants; Asthma; Cells, Cultured; Disease Models, Animal; Epithelial Cells; Epithelial-Mesenchymal Transition; Glucosides; Humans; Lung; Male; Mice; Mice, Inbred BALB C; Mice, Nude; NF-E2-Related Factor 2; Ovalbumin; Reactive Oxygen Species; Signal Transduction; Stilbenes; Transforming Growth Factor beta1

Sufficient exposure to natural environments, in particular soil and its microbes, has been suggested to be protective against allergies.. We aim at gaining more direct evidence of the environment-microbiota-health axis by studying the colonization of gut microbiota in mice after exposure to soil and by examining immune status in both a steady-state situation and during allergic inflammation.. The gastrointestinal microbiota of mice housed on clean bedding or in contact with soil was analyzed by using 16S rRNA gene sequencing, and the data were combined with immune parameters measured in the gut mucosa, lung tissue, and serum samples.. We observed marked differences in the small intestinal and fecal microbiota composition between mice housed on clean bedding or in contact with soil, with a higher proportion of Bacteroidetes relative to Firmicutes in the soil group. The housing environment also influenced mouse intestinal gene expression, as shown by upregulated expression of the immunoregulatory markers IL-10, forkhead box P3, and cytotoxic T lymphocyte-associated protein 4 in the soil group. Importantly, using the murine asthma model, we found that exposure to soil polarizes the immune system toward T. Our results provide evidence of the role of environmentally acquired microbes in alleviating against T

Topics: Allergens; Animals; Asthma; Cytokines; Disease Models, Animal; Feces; Female; Gastrointestinal Microbiome; Immune Tolerance; Intestine, Small; Mice, Inbred BALB C; Ovalbumin; RNA, Ribosomal, 16S; Soil; Soil Microbiology

Asthma is characterized by chronic inflammation, and long-term chronic inflammation leads to airway remodeling. But the potential regulatory mechanism of airway remodeling is not clearly understood, and there is still no effective way to prevent airway remodeling. Present studies have confirmed the role of microRNAs (miRNAs) in the development of disease, which is known as suppressing translation or degradation of messenger RNA (mRNA) at the posttranscriptional stage. In this study, we described the role of miRNA-133a in asthma and demonstrated it in regulating airway remodeling of asthma through the phosphoinositide 3 kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway by targeting IGF-1 receptor (IGF1R). IGF1R helps in mediating the intracellular signaling cascades. Asthmatic mice models were established by sensitization and Ovalbumin challenge. Adenovirus transfer vector carrying miR-133a or miR-133a sponge sequence was used to build the overexpression or downexpression of miR-133a modeling. Real-time polymerase chain reaction and Western blot were used to determine the alterations in the expression of miR-133a and mRNAs and their corresponding proteins. Results showed that miR-133a was downregulated in asthma. Upregulation of miR-133a expression in airway smooth muscle cells in vivo and in vitro could inhibit the activation of PI3K/AKT/mTOR pathway, and reduce the expression of α-smooth muscle actin (α-SMA), indicating that airway remodeling was inhibited. Functional studies based on luciferase reporter revealed miR-133a as a direct target of IGF1R mRNA. In conclusion, these data suggested that miR-133a regulated the expression of α-SMA through PI3K/AKT/mTOR signaling by targeting IGF1R. miR-133a plays a key role in airway remodeling of asthma and may serve as a potential therapeutic target for managing asthmatic airway remodeling.

Topics: Actins; Airway Remodeling; Airway Resistance; Animals; Asthma; Cells, Cultured; Disease Models, Animal; Female; Gene Expression Regulation; Lung; Mice, Inbred BALB C; MicroRNAs; Myocytes, Smooth Muscle; Ovalbumin; Phosphatidylinositol 3-Kinase; Proto-Oncogene Proteins c-akt; Receptor, IGF Type 1; Signal Transduction; TOR Serine-Threonine Kinases

Pingchuanning decoction is a well-known traditional Chinese medicine for the treatment of airway inflammatory diseases, including asthma. However, the potential mechanism by which Pingchuanning decoction contributes to the amelioration of airway inflammation remains unknown.. A rat model of asthma was well established by inducing ovalbumin. Lipopolysaccharide-stimulated rat tracheal epithelial (RTE) cells were used as cellular model. Lung histopathology and goblet cell hyperplasia were assessed by hematoxylin-eosin (HE) and periodic acid Schiff staining, respectively. Total inflammatory cells count and RTE cell apoptosis were analyzed by flow cytometry. The autophagic activities were evaluated by immunohistochemical and immunofluorescence analysis and Western blot analysis of autophagy-related proteins. We also detected the effects of Pingchuanning decoction on phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) and high-mobility group box 1 (HMGB1)-mediated toll-like receptor 4 (TLR4)/NF-κB pathways-related proteins and inflammatory cytokines using the Western blot analysis and enzyme-linked immunosorbent assay.. Pingchuanning decoction effectively attenuated pulmonary pathology and autophagy. Treatment with Pingchuanning decoction activated PI3K/Akt/mTOR pathway and inhibited HMGB1/TLR4/NF-κB pathway, which could be overturned by LY294002, a PI3K antagonist, or rapamycin (Rapa), an autophagy inducer.. Pingchuanning decoction exerted a therapeutic effect on asthma by inhibiting autophagy via PI3K/Akt /mTOR signaling pathway.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Autophagy; Dexamethasone; Disease Models, Animal; Drugs, Chinese Herbal; Epithelial Cells; Gene Expression Regulation; Lung; Male; Ovalbumin; Phosphatidylinositol 3-Kinase; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Respiratory Mucosa; Signal Transduction; TOR Serine-Threonine Kinases; Treatment Outcome

The asthma candidate gene inositol polyphosphate 4-phosphatase type I A (INPP4A) is a lipid phosphatase that negatively regulates the PI3K/Akt pathway. Destabilizing genetic variants of INPP4A increase the risk of asthma, and lung-specific INPP4A knockdown induces asthma-like features. INPP4A is known to localize intracellularly, and its extracellular presence has not been reported yet. Here we show for the first time that INPP4A is secreted by airway epithelial cells and that extracellular INPP4A critically inhibits airway inflammation and remodeling. INPP4A was present in blood and BAL fluid, and this extracellular INPP4A was reduced in patients with asthma and mice with allergic airway inflammation. In both naive mice and mice with allergic airway inflammation, antibody-mediated neutralization of extracellular INPP4A potentiated PI3K/Akt signaling and induced airway hyperresponsiveness, with prominent airway remodeling, subepithelial fibroblast proliferation, and collagen deposition. The link between extracellular INPP4A and fibroblasts was investigated in vitro. Cultured airway epithelial cells secreted enzymatically active INPP4A in extracellular vesicles and in a free form. Extracellular vesicle-mediated transfer of labeled INPP4A, from epithelial cells to fibroblasts, was observed. Inhibition of such transfer by anti-INPP4A antibody increased fibroblast proliferation. We propose that secretory INPP4A is a novel "paracrine" layer of the intricate regulation of lung homeostasis, by which airway epithelium dampens PI3K/Akt signaling in inflammatory cells or local fibroblasts, thereby limiting inflammation and remodeling.

Topics: Airway Remodeling; Animals; Asthma; Cell Line, Transformed; Cell Line, Tumor; Cell Proliferation; Disease Models, Animal; Fibroblasts; Humans; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphatidylinositol 3-Kinases; Phosphoric Monoester Hydrolases; Respiratory Hypersensitivity; Signal Transduction

Although excitements related to stem cell therapeutic outcomes have been highlighted enormously in asthma, the vast majority of works were conducted by researchers in animal models. Elucidating the mechanisms underlying the therapeutic effects of MSCs in asthmatic rats will provide a rational basis for assuring maximal safety of future clinical application of stem cells. In the current study, we sought to investigate the possible paracrine mechanism by which direct injection of MSCs and/or CM attenuate efficiently Th2-mediated inflammation in asthmatic lung tissues with the focus on ICAM-1 and VCAM-1 expression.. Male rats were divided into four experimental groups (n = 6); healthy rats received PBS intratracheally (group C), sensitized rats received PBS intratracheally (group S), sensitized rats received CM intratracheally (group S + CM), and sensitized rats received PBS intratracheally containing 2 × 10. Our results showed CM, and notably rBMMSCs, returned the expression of IL-5, IL-12, INF-γ, ICAM-1, and VCAM-1 (p < 0.001 to p < 0.05) to the normal levels. Based on data, pathological injuries in pulmonary specimens of asthmatic rats were significantly attenuated (p < 0.001 to p < 0.05). Moreover, rBMMSCs had potential to successfully home to an asthmatic niche in cell-administrated rats.. Our data noted the potency of CM and especially MSCs in ameliorating pathological changes via intra-tracheal route presumably by targeting ICAM-1 and VCAM-1 in lung tissues in rat asthma model.

Topics: Animals; Asthma; Bone Marrow Cells; Bone Marrow Transplantation; Cell Adhesion Molecules; Cells, Cultured; Culture Media, Conditioned; Cytokines; Disease Models, Animal; Endothelial Cells; Inflammation Mediators; Intercellular Adhesion Molecule-1; Lung; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Ovalbumin; Paracrine Communication; Phenotype; Rats, Wistar; Stem Cell Niche; Th2 Cells; Vascular Cell Adhesion Molecule-1

Involucrum castaneae(IC)is used in Chinese folk medicine to treat various lung diseases, as well as for its reducing phlegm and anti-inflammatory properties.. The purpose of this experiment is to verify the effect of IC on airway inflammation, responsiveness in ovalbumin (OVA)-induced asthmatic guinea pigs. The main chemical components of IC were also analyzed.. The potential of the ethanol extract of Involucrum castaneae (EEIC) to protect against OVA-induced allergic airway response in guinea pigs was investigated. The latency of asthma in guinea pigs were recorded after the allergic asthma induced. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of immunoglobulin E (IgE), interleukin-5 (IL-5), nerve growth factor (NGF) and interferon-γ (IFN-γ) in asthma allergy. Reverse transcription-PCR (RT-PCR) was used to detect the expression of IL-5 mRNA in asthmatic guinea pig lungs. Paraffin sections of lung tissue were used to analyze pathological changes. The total flavonoid content was determined and the chemical components were analyzed by LC-MS/MS.. It was found that EEIC was able to reduce the number of eosinophil (EOS) in bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) in the guinea pig model of OVA -induced asthma. Meanwhile, it also significantly reduced the levels of inflammation-related factors IgE and IL-5, decreased the expression of IL-5 mRNA in lung tissue, and increased the level of IFN-γ. Pathological examination of paraffin section of lung tissue showed that EEIC can reduce the thickening of bronchial smooth muscle and reduce the infiltration damage of tissues by various inflammatory cells. The presence of flavonoids, terpenoids and phenolic compounds in EEIC might be responsible for these activities.. IC alleviated airway inflammation and smooth muscle thickening in guinea pigs with OVA-sensitized allergic asthma. The paper explains the traditional efficacy and material basis of IC and lays a foundation for further development.

Topics: Airway Remodeling; Allergens; Animals; Anti-Asthmatic Agents; Asthma; Ethanol; Fagaceae; Guinea Pigs; Immunoglobulin E; Interferon-gamma; Interleukin-5; Lung; Male; Muscle, Smooth; Ovalbumin; Plant Extracts; Solvents

MicroRNAs (miRNAs) are small yet versatile gene tuners that regulate a variety of cellular processes, including cell growth and proliferation. The aim of this study was to explore how miR-448-5p affects airway remodeling and transforming growth factor-β1 (TGF-β1)-stimulated epithelial-mesenchymal transition (EMT) by targeting Sine oculis homeobox homolog 1 (Six1) in asthma. Asthmatic mice models with airway remodeling were induced with ovalbumin solution. MiRNA expression was evaluated using quantitative real-time polymerase chain reaction. Transfection studies of bronchial epithelial cells were performed to determine the target genes. A luciferase reporter assay system was applied to identify whether Six1 is a target gene of miR-448-5p. In the current study, we found that miR-448-5p was dramatically decreased in lung tissues of asthmatic mice and TGF-β1-stimulated bronchial epithelial cells. In addition, the decreased level of miR-448-5p was closely associated with the increased expression of Six1. Overexpression of miR-448-5p decreased Six1 expression and, in turn, suppressed TGF-β1-mediated EMT and fibrosis. Next, we predicted that Six1 was a potential target gene of miR-448-5p and demonstrated that miR-448-5p could directly target Six1. An SiRNA targeting Six1 was sufficient to suppress TGF-β1-induced EMT and fibrosis in 16HBE cells. Furthermore, the overexpression of Six1 partially reversed the protective effect of miR-448-5p on TGF-β1-mediated EMT and fibrosis in bronchial epithelial cells. Taken together, the miR-448-5p/TGF-β1/Six1 link may play roles in the progression of EMT and pulmonary fibrosis in asthma.

Topics: Animals; Asthma; Cell Line; Epithelial Cells; Epithelial-Mesenchymal Transition; Female; Fibrosis; Gene Knockdown Techniques; Homeodomain Proteins; Humans; Mice; MicroRNAs; Ovalbumin; Random Allocation; Respiratory Mucosa; Transforming Growth Factor beta1

Interleukin-27 (IL-27) modulates CD4+ T-cell differentiation and function. The aim of this study is to investigate the effect and molecular mechanisms of IL-27 on the development of asthma.. IL-27 was intranasally administered in an ovalbumin-induced asthma model, and lung mononuclear cells and different Th cell classes were detected by fluorescence-activated cell sorting. The effect and mechanisms of IL-27 on human bronchial epithelial (HBE) cells were investigated by measuring changes in chemotactic factors, cytokines, transcription factors, and signaling pathways.. We found that intranasal administration of IL-27 could attenuate airway inflammation and hyperresponsiveness, upregulate the type 1 T helper (Th1)-T memory (Tm) cells and regulatory T (Treg) cells subgroups of lung tissue lymphocytes, and diminish the levels of type 2 T helper (Th2) cytokines. IL-27 upregulated the expression of C-C motif chemokine ligand 2 (CCL2), CCL3, and CCL4 in HBE cells and promoted the production of chemotactic factors to attract monocyte recruitment. Recruited monocytes secondarily secreted IL-27 to influence HBE cells in a positive feedback cycle. After IL-27 intervention, signal transducer and activator of transcription 1 (STAT1) phosphorylation increased, while STAT4 and STAT6 phosphorylation declined.. Preventative intranasal administration of IL-27 can recruit more IL-27-secreted monocytes to the airway and change the different T-cell classes in lung. The improved Th1 environment helps to alleviate Th2-mediated allergic asthma by repairing the STAT1 pathway but not the STAT4 pathway.

Topics: Animals; Asthma; Cytokines; Female; Interleukin-27; Interleukin-4; Lung; Mice, Inbred C57BL; Ovalbumin; STAT6 Transcription Factor; Th2 Cells

Th1, Th2, Th17, and Treg cells and their cytokine gene expressions in splenocytes of control mice, ovalbumin sensitized (S), and S treated with dexamethasone and carvacrol during a sensitization period were examined. Th2 and Th17 population as well as the gene expression of IL-4, IL-17, and TGF-β were increased, but Th1, Th1/Th2 ratio, the gene expression of IFN-γ and FOXP3 as well as the IFN-γ/IL-4 ratio were decreased in S compared with control group ( P < 0.001 for all cases). Carvacrol treatment caused significant reduction of Th2 and Th17 population as well as gene expression of IL-4, IL-17, and TGF-β but increase in Treg cells, Th1/Th2 ratio, gene expressions of FOXP3, IFN-γ, and IFN-γ/IL-4 ratio ( P < 0.05 to P < 0.001). The population of Th1, Th2, Th17 cells as well as the gene expression of IL-4, IL-17, and TGF-β were significantly decreased, but only Treg was increased in the dexamethasone treatment group ( P < 0.05 to P < 0.001). Carvacrol treatment during the sensitization period showed a more specific effect on Th1/Th2 imbalance in sensitized mice than dexamethasone, which may indicate the therapeutic potentials of carvacrol in disorders associated with Th1/Th2 imbalance such as asthma.

Topics: Animals; Asthma; Cymenes; Cytokines; Dexamethasone; Gene Expression; Gene Expression Regulation; Humans; Interferon-gamma; Interleukin-17; Interleukin-4; Mice; Ovalbumin; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Th2 Cells; Transforming Growth Factor beta

Previously, we described tracheal rat rings relaxation by several flavonoids, being 6-hydroxyflavone (6-HOF) the most active derivative of the series. Thus, its mechanism of action was determined in an ex vivo tracheal rat ring bioassay. The anti-asthmatic effect was assayed in in vivo OVAlbumin (OVA)-sensitized guinea pigs. Finally, the toxicological profile of 6-HOF was studied based on Organization of Economic Cooperation and Development guidelines with modifications. 6-HOF-induced relaxation appears to be related with receptor-operated calcium channel and voltage-operated calcium channel blockade as the main mechanism of action, and also through the production of relaxant second messengers NO and cGMP. Molecular docking supports that 6-HOF acts as calcium channel blocker and by activation of nitric oxide synthase. In addition, the in vivo anti-asthmatic experiments demonstrate the dose-dependent significant anti-allergic effect of 6-HOF induced by OVA, with best activity at 50 /kg. Finally, toxicological studies determined a LD50 > 2,000 mg/kg and, after 28 day of treatment with 6-HOF (50 mg/kg) by intragastric route, mice did not exhibit evidence of any significant toxicity. In conclusion, experiments showed that 6-HOF exerts significant relaxant activity through calcium channel blockade, and possibly, by NO/cGMP-system stimulation on rat trachea, which interferes with the contraction mechanism of smooth muscle cells in the airways. In addition, the flavonoid shows potential anti-asthmatic properties in an anti-allergic pathway. Furthermore, because the pharmacological and safety evidence, we propose this flavonoid as lead for the development of a novel therapeutic agent for the treatment of asthma and related respiratory diseases.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Asthma; Calcium Channels, L-Type; Flavonoids; Guinea Pigs; In Vitro Techniques; Male; Mice, Inbred ICR; Molecular Docking Simulation; Muscle Relaxation; Muscle, Smooth; Nitric Oxide Synthase Type III; Ovalbumin; Rats, Wistar; Toxicity Tests; Trachea

Asthma is a chronic allergic ailment affecting a considerably large population of the world. The aim of this study is to evaluate the ameliorative effects of vanillic acid against ovalbumin (OVA)-induced asthma in rat model. Asthma was induced in Sprague Dawley rats and vanillic acid was orally administered at 25 and 50 mg/kg body weight for 28 days. Rats challenged with OVA showed heavy signs of airway inflammation and remodeling similar to chronic asthma, evidenced by the increased differential cell counts and presence of inflammatory cytokines in the bronchoalveolar lavage fluid (BALF), along with elevated serum immunoglobulin levels, and the histological results. However, vanillic acid dose-dependently attenuated the manifestation of OVA-induced asthma (p < 0.05) through suppression of inflammatory mediators and modulation of immunoglobulin levels in rats. The asthma mitigating properties of vanillic acid might be due to suppression of oxidative stress and prevention of lung airway inflammation.

Topics: Animals; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Immunoglobulin E; Inflammation; Lung; Male; Malondialdehyde; Organ Size; Ovalbumin; Rats; Rats, Sprague-Dawley; Vanillic Acid

Topics: Airway Remodeling; Asthma; Autophagy; Humans; Ovalbumin; Respiratory System

Studies from epidemiology suggest that ambient temperature is one of the underlying triggers and potential causes of asthma. The aim of this study was to examine the impact and the molecular mechanism of temperature-invoked airway inflammation using an experimental model of asthma in BALB/c mice.. Mice were exposed to different temperature conditions (steady 26°C, 26°C/18°C cycle, 26°C/10°C cycle) and received sensitization and challenge of ovalbumin (OVA) during a 21-day period. HC030031, a selective transient receptor potential A1 (TRPA1) channel blocker, was used to investigate the underlying mechanism of TRPA1 in 'asthmatic' airways. After the final OVA challenge, in vivo lung function was measured, and bronchoalveolar lavage fluid (BALF) and pulmonary inflammation were assessed.. The temperature variations, especially the largest temperature difference (16°C), exacerbated airway inflammation in OVA-induced mice, increasing the levels of serum total-IgE (immunoglobulin E) and IgG1, inflammatory cells and cytokines in BALF. Analysis of histopathological changes and lung function verified that repeated exposure to very cold and changed temperatures aggravated airway hyperresponsiveness (AHR). Significant upregulation of TRPA1 expression was revealed by immunohistochemistry in the presence of the largest temperature variation (26°C/10°C cycle), while administration of HC030031 successfully inhibited TRPA1 expression, thus attenuating the asthma-like pathological features.. Repeated exposure to temperature variation exacerbated experimental 'asthma' and TRPA1 mediated this temperature-dependent inflammatory effect.

Topics: Acetanilides; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Immunoglobulin E; Immunoglobulin G; Inflammation; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Purines; Temperature; TRPA1 Cation Channel

Vitamin D produces an anti-allergic effect that prevents inflammation due to asthma.. We investigated whether vitamin D has an anti-inflammatory effect on the sensitization and challenge stages of asthma development in a murine model.. Mice were divided into the following five groups according to ovalbumin (OVA) and vitamin D (VD) administration: control group, OVA group, preventive VD group (VD injection before OVA sensitization), inhibitory VD group (VD injection after OVA challenge), and dual VD groups (VD injection before OVA sensitization and after OVA challenge). Each group was evaluated for airway hyperresponsiveness (AHR), cell counts, cytokines, total IgE, and OVA-specific IgE using bronchoalveolar lavage fluid (BALF). Cytokines in the lysate and eosinophils in the lung tissue were also evaluated.. AHR occurred less in the groups to which VD was administered than in the OVA group. The eosinophils, neutrophils, IL-5 in BALF, IL-4, TGF-β, and eosinophils in lysate decreased with the administration of VD in the preventive, inhibitory, and dual VD groups compared with the OVA group. Although the lymphocytes, macrophages, IL-4 in BALF, and IL-5 in lysate decreased with administration of VD in the inhibitory and dual VD groups, they were not affected by preventive VD administration. These anti-allergic effects of VD were most noticeable with VD administration for dual (preventive and inhibitory) purposes.. VD may produce preventive and inhibitory effects on the development and exacerbation of asthma in a murine model. These effects are most noticeable when VD is used for dual purposes.

Topics: Allergens; Animals; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Immunoglobulin E; Immunomodulation; Inflammation Mediators; Mice; Ovalbumin; Vitamin D

Ginsenoside Compound K (CK) showed potent activity against IgE for the treatment of asthma. A series of CK analogues were then synthesized by straightforward procedures. The in vivo anti-IgE activity evaluations using the OVA-induced asthmatic mouse model revealed preliminary SARs of the CK analogues, which showed that the sugar type, modifications on A-ring and the C20 side chain of CK all affected much on the activities. Primary SARs optimization led to the discovery of compounds T1, T2, T3, T8 and T12, which displayed superior or comparable anti-asthmatic effects (IgE value = 1237.11 ± 106.28, 975.82 ± 160.32, 1136.96 ± 121.85, 1191.08 ± 107.59 and 1258.27 ± 148.70 ng/mL, respectively) in comparison with CK (1501.85 ± 184.66 ng/mL). These potent compounds could serve as leads for further development.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Disease Models, Animal; Dose-Response Relationship, Drug; Ginsenosides; Immunoglobulin E; Mice; Molecular Conformation; Ovalbumin; Structure-Activity Relationship

Thymic stromal lymphopoietin (TSLP) mediates immune reaction in patients with asthma. Matrix metalloproteinase (MMP), connective tissue growth factor (CTGF), and transforming growth factor-β (TGF-β) are inflammatory mediators whose responses to the anti-TSLP antibody are unknown. This study examined the effect of an anti-TSLP antibody on MMP, CTGF, TGF-β, and airway structural changes in airway remodeling in asthma.. Mice were randomly divided into phosphate-buffered-saline-challenged (PBS), ovalbumin-challenged (OVA), and ovalbumin-challenged with anti-TSLP antibody (OVA + anti-TSLP) groups. Airway responsiveness and serum ovalbumin-specific immunoglobulin E were measured. Differential cell counts and MMP-2 and MMP-9 were evaluated in bronchoalveolar lavage fluid (BALF). Airway structural changes were quantified using morphometric analysis and presentation by immunohistochemistry staining. Lung CTGF, TGF-β, and TSLP were analyzed using western blot.. Airway responsiveness was significantly lower in OVA + anti-TSLP and PBS groups than in OVA group. Airway structural changes exhibited less smooth muscle thickness in OVA + anti-TSLP and PBS groups than in OVA group. MMP-2 and MMP-9 in BALF and CTGF, TGF-β, and TSLP in lungs significantly decreased in OVA + anti-TSLP and PBS groups compared with OVA group.. Anti-TSLP antibody exerts the preventive effect of decreasing airway structural changes through reduction of MMP, TGF-β, and CTGF in airway remodeling of asthma.

Topics: Airway Remodeling; Animals; Antibodies, Monoclonal; Asthma; Connective Tissue Growth Factor; Cytokines; Disease Models, Animal; Female; Immunoglobulin E; Inflammation; Lung; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Matrix Metalloproteinases; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Plethysmography; Thymic Stromal Lymphopoietin; Transforming Growth Factor beta

The incidence and severity of asthma preponderate in women versus men. Leukotrienes (LTs) are lipid mediators involved in asthma pathogenesis, and sex disparities in LT biosynthesis and anti-LT pharmacology in inflammation have recently emerged. Here, we report on sex dimorphism in LT production during allergen sensitization and its correlation to lung function. While high plasma levels of IgE, as sensitization index, were elevated in both sexes, LT levels increased only in lungs of female ovalbumin-sensitized BALB/c mice. Sex-dependent elevated LT levels strictly correlated to an enhanced airway hyperreactivity, pulmonary inflammation and mast cell infiltration/activation in female mice. Importantly, this sex bias was coupled to superior therapeutic efficacy of different types of clinically used LT modifiers like zileuton, MK886 and montelukast in female animals. Our findings reveal sex-dependent LT production as a basic mechanism of sex dimorphism in allergic asthma, and suggest that women might benefit more from anti-LT asthma therapy.

Topics: Allergens; Animals; Asthma; Female; Immunoglobulin E; Leukotrienes; Lung; Male; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Sex Characteristics

Yan-Hou-Qing (YHQ), a Chinese medicine formula containing fourteen kinds of materials, has been designed for pharyngitis and cough treatment in Oriental medicine. In the present study, the anti-allergic effects and underlying mechanisms of YHQ in inhibition of airway hyper responsiveness (AHR) was explored in an ovalbumin (OVA)-induced allergic asthma mouse model.. BALB/c mice were sensitized by OVA and cholera toxin (CT) and challenged with OVA intranasally to induce allergic asthma mouse model. YHQ (200 mg/kg) was orally administered for 3 weeks from week-2 after OVA sensitization. The AHR and histological changes of lung tissues were evaluated by whole-body barometric plethysmography analysis and hematoxylin and eosin (H&E) staining, respectively. The serum concentration of OVA-specific IgE and T helper 2 (Th2) cytokines (IL-4 and IL-13) were determined by enzyme-linked immune sorbent assay (ELISA). Flow cytometry was performed to evaluate the percentage of CD4. The elevated AHR responses, heavier inflammatory cell infiltration and Th2 cytokines in allergic asthma group indicated Ovalbumin-induced asthmatic mouse models were built successfully. Compared to allergic asthma group, OVA-induced AHR responses and eosinophil infiltration in lung were improved significantly, and the productions of OVA-specific IgE and Th2 cytokines, IL-4 and IL-13, in the serum were also reduced dramatically after the treatment of YHQ. Moreover, YHQ treatment significantly increased the percentage of CD4. YHQ improves the allergic asthma related symptoms via promotion of CD4

Topics: Allergens; Animals; Anti-Asthmatic Agents; Asthma; Cholera Toxin; Disease Models, Animal; Drugs, Chinese Herbal; Female; Immunoglobulin E; Interleukin-13; Interleukin-4; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Th2 Cells

Zizyphus jujuba Mill, a famous oriental traditional medicine, has been reported to exhibit diverse activities in biological systems including the respiratory system. However, a little information is available on its antiasthmatic activity. Jujuboside B (JB) is a natural saponin and one of the active constituent of fruits of Zizyphus jujuba. In the present investigation, JB was isolated from ethanolic extracts of fruits of Zizyphus jujuba (EZJF). EZJF and JB were then evaluated for anti-asthmatic activity using various screening methods. JB was additionally evaluated using ovalbumin (OVA) -induced allergic asthma in mice. Results obtained in the present study showed that EZJF and JB significantly inhibited clonidine-induced catalepsy, milk-induced leucocytosis and eosinophilia, clonidine-induced mast cell degranulation, and passive paw anaphylaxis. The number of inflammatory cells in bronchoalveolar lavage (BAL) fluid was considerably lowered and the severity of pulmonary inflammation was alleviated in the mice pretreated with JB. The high-level expression of T-helper type 2 (TH2) cytokines was markedly reduced in the serum, BAL fluid, and lung homogenates. Thus EZJF and JB showed potent anti-asthmatic activity. Hence EZJF and JB possess a potential role in the treatment of asthma.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Catalepsy; Clonidine; Cytokines; Disease Models, Animal; Eosinophilia; Leukocytosis; Lung; Mast Cells; Medicine, Chinese Traditional; Mice; Milk; Ovalbumin; Plant Extracts; Rats; Rats, Wistar; Saponins; Ziziphus

Asthma is a common chronic inflammatory respiratory disease characterised by airway inflammation and hyperresponsiveness. The present study was designed to clarify the effect of intranasal miR‑410 administration in an ovalbumin (OVA)‑induced murine model of asthma. It was found that miR‑410 expression was significantly decreased in the lungs of OVA‑induced asthmatic mice (P<0.05) and miR‑410 was overexpressed via intranasal instillation. Bioinformatics indicated that the 3'‑untranslated regions of interleukin (IL)‑4 and IL‑13 messenger RNAs (mRNAs) contain miR‑410 binding sites. The IL‑4 and IL‑13 genes were confirmed to be miR‑410‑regulated using the dual‑luciferase reporter assay. Additionally, intranasal administration of miR‑410 markedly attenuated airway inflammation and reduced infiltration of inflammatory cells into bronchoalveolar lavage fluid (P<0.05) as determined by bronchoalveolar lavage fluid analysis. Moreover, miR‑410 significantly decreased the lung expression of IL‑4 and IL‑13 (P<0.05), although the levels of mRNAs encoding IL‑4 and IL‑13 in lungs did not change significantly as determined by real‑time PCR analysis. In conclusion, we found that intranasal administration of miR‑410 effectively inhibited airway inflammation in OVA‑induced asthmatic mice by targeting IL‑4 and IL‑13 at the post‑transcriptional level. miR‑410 is thus a promising treatment for allergic asthma.

Topics: 3' Untranslated Regions; Administration, Intranasal; Animals; Asthma; Bronchoalveolar Lavage Fluid; Female; Inflammation; Interleukin-13; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin

Asthma is a worldwide public health issue, affecting the sufferer's quality of life. Many researchers are extensively studying the cellular processes involved in the affected airways. Experimental asthma using animals has been performed for a long time, mainly applying murine models due to well-known advantages. The aim of this study is to present an allergic airway inflammation protocol in mice. Basically, the allergic airway inflammation is induced by intraperitoneal sensitization and intratracheal challenge with ovalbumin (OVA). The model provided here mimics acute asthma characteristics including excessive mucus production, airway hyperresponsiveness, and eosinophilic airway inflammation.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity

An animal (BALB/c mice) model of catalpol associated with bronchial asthma in vivo was established, and the effects of catalpol and its relationship with cytokines were investigated. A total of 30 adult BALB/c mice were randomly divided into a positive control group, a model group, and a catalpol group, with 10 mice in each group. The lung function of mice, the cell count, and the cytokine concentrations in bronchoalveolar lavage fluid (BALF) were detected. The levels of cytokines [interleukin 4 (IL-4), interleukin 5 (IL5), and interferon gamma (IFN-γ)] in BALF were measured with enzyme-linked immunosorbent assay methods. The total number of cells in the BALF of the group treated with catalpol was significantly lower than the model group. After treatment with catalpol, the eosinophils and neutrophils of the mice were remarkably reduced compared with the model group. The malondialdehyde content in the lung tissue homogenate of the mice was also decreased in the catalpol group. The cytokines IL-5 and IL-4 exhibited a similar tendency: the concentrations of IL-4 and IL-5 for the catalpol group were dramatically decreased compared with the model group. However, the IFN-γ concentration for the catalpol group was higher than the model group. The results indicated that IL-5 may involve in the pathologic process of asthma-like IL-4, and an inflammatory reaction may still exist in the airway during the remission stage of asthma. The imbalances of the cytokine network might be an important molecular basis in the asthma pathogenesis. It is suggested that catalpol may be a potential drug for the clinical treatment of asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Gene Expression Regulation; Interferon-gamma; Interleukin-4; Interleukin-5; Iridoid Glucosides; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Random Allocation; Up-Regulation

Natural killer T cells (NKT cells) and regulatory T cells (Treg cells) are two important immune regulatory cells which both play critical roles in asthma. Our previous experiments revealed that activation of Treg cells suppressed NKT cells in asthma. However, the possible regulatory effects and the mechanisms linking Treg cells and NKT cells remain poorly understood. The current study was designed to further investigate the regulatory effect and its possible mechanisms of Treg cells on NKT cells function, especially the distribution of NKT cells. Regulatory T cell (Treg), responder T cell (Teff) and Natural killer T cell (NKT) were isolated and purified. After Lentivirus carrying CD39 (Le-CD39) was transfected into Treg cells, the immune phenotype of Treg cells was changed and the suppressive effect of Treg cells on Teff cells with an activation of Treg cells was enhanced, marking with a high expression level of interleukin 10 (IL-10) and transforming growth factor β (TGF-β). Up-regulation of CD39 expression led to lower ATP level in cell culture supernatant. To further explore its function in asthma, we introduced an ovalbumin (OVA)-induced mice model of asthma. And the data showed that up-regulation of CD39 remarkably alleviated OVA-induced hallmarks of the asthma and increased NKT cells in the spleen and peripheral blood; however, decreased NKT cells in the lung. Furthermore, up-regulation of CD39 decreased the levels of interleukin 4 (IL-4) and interferon γ (IFN-γ) in the lung of OVA-treated mice. Our results strongly suggest that Treg cells could be activated by CD39 signal transduction and then affected the distribution of NKT cells in the OVA-induced mice model of asthma.

Topics: Adenosine Triphosphate; Animals; Antigens, CD; Apyrase; Asthma; Cells, Cultured; Disease Models, Animal; Gene Expression; Interleukin-10; Male; Mice, Inbred BALB C; Natural Killer T-Cells; Ovalbumin; Signal Transduction; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Up-Regulation

Anthriscus sylvestris L. Hoffmann (AS) is a perennial plant that grows in Asia and Eastern Europe. Its dried root is used to treat conditions such as asthma, bronchitis, and cough.. The present study investigated the anti-inflammatory effects of whole AS extract (ASE) on allergic lung inflammation in vitro and in vivo as well as the underlying mechanisms.. We used an ovalbumin (OVA)-induced asthma mouse model and in vitro primary T helper (Th)2 polarization system. Five groups of 8-week-old female C57BL/6 mice were divided into the following groups: saline control, or OVA-induced allergic asthma with vehicle, ASE (100 or 200 mg/kg), or dexamethasone (5 mg/kg) treatment for 7 days.. ASE attenuated mucus secretion in airway epithelial cells, inflammatory cell infiltration, eosinophilia, and Th2 cytokine levels in bronchoalveolar lavage fluid. Mice administered ASE showed reductions in the activated cluster of differentiation 4. ASE exerts anti-asthmatic effects by inhibiting IRF4 expression and thereby suppressing Th2 cell activation.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Apiaceae; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; Interferon Regulatory Factors; Mice; Mice, Inbred C57BL; Ovalbumin; Plant Extracts; Plant Roots; RAW 264.7 Cells; Th2 Cells

Acid sphingomyelinase (ASM) is one of the enzymes that catalyzes the breakdown of sphingomyelin to ceramide and phosphorylcholine. In this study, we aimed at elucidating the role of ASM in allergic asthma. We used an ovalbumin-induced murine model of asthma where we compared wild-type and ASM-deficient mice. In wild-type mice, secretory ASM activity in the bronchoalveolar lavage fluid was increased in the acute ovalbumin model, but not in a tolerogenic model. Furthermore, in the absence of ASM, the serum IgE level was reduced, compared with wild-type mice, while an accumulation of interstitial macrophages and foreign antigen-induced regulatory T cells along with exhausted CD4

Topics: Animals; Asthma; Disease Models, Animal; Female; Immunoglobulin E; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Sphingomyelin Phosphodiesterase; T-Lymphocytes, Regulatory

Allergic asthma and atherosclerosis represent different directions of inflammatory responses of CD4. We aimed to investigate the roles of curcumin in asthma-accelerated atherosclerosis plaque formation, and the change of CD4. Six to eight-week-old apolipoprotein E. The accelerated atherosclerosis was induced by allergic asthma accompanied by increased T helper cell (Th)2 and Th17 cells and decreased regulatory T cells (Tregs) in the spleen. After the 8-week treatment with curcumin, the lesion areas in the aortic root in asthmatic mice significantly improved, and the elevated Th2 and Th17 cells significantly decreased, but Tregs markedly increased. Although curcumin treatment markedly reduced the interleukin (IL)-4 and IL-13 in serum and spleen, the elevated IL-17A did not decrease. Moreover, Th1 cells showed no significant change between different groups. The mRNA expression levels of M1 macrophage-related inflammatory factors IL-6, iNOS and IL-1β were markedly elevated in the spleens of asthmatic mice, but significantly decreased after the 8-week treatment with curcumin.. Curcumin ameliorated the aggravation of atherosclerotic lesions and stabilised plaque by modulating the balance of Th2/Tregs in asthmatic apoE

Topics: Animals; Asthma; Atherosclerosis; Curcumin; Interleukins; Mice; Mice, Inbred C57BL; Mice, Knockout, ApoE; Ovalbumin; Plaque, Atherosclerotic; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Th2 Cells

Allergic asthma is a chronic immune-inflammatory disorder, characterized by airway inflammation and airway hyperresponsiveness (AHR). Morin is a natural flavonoid reported to exhibit inhibitory action against IgE-mediated allergic response.. To determine the efficacy of murine model of ovalbumin (OVA)-induced AHR inhibition by morin and decipher the molecular mechanism involved.. Sprague-Dawley rats were sensitized and challenged with OVA to induce AHR. Rats received treatment with morin (10, 30 and 100 mg/kg, p.o.) for the next 28 days.. Morin (30 and 100 mg/kg) significantly and dose-dependently attenuated (p < 0.01 and p < 0.001) OVA-induced alterations in pulse oxy and lung function test, increased bronchoalveolar lavage fluid cell counts, elevated total protein and albumin levels in serum, BALF, and lungs, increased serum total and OVA-specific IgE levels and, elevated oxidative stress levels in the lung. RT-PCR analysis revealed that morin treatment (30 and 100 mg/kg) significantly (p < 0.001) up-regulated SUMF2 mRNA expression in lungs whereas mRNA expressions of BLT2, NF-κB, and Th2-cytokine (TNF-α, IL-1β, IL-4, IL-6, and IL-13) were down-regulated significantly and dose-dependently (p < 0.01 and p < 0.001). Also, histologic and ultrastructural studies showed that morin significantly inhibited (p < 0.001) OVAinduced perivascular and peribranchial inflammatory infiltration and interstitial fibrosis.. Morin exhibited inhibitory effect against OVA-induced allergic asthma by activation of SUMF2 which impeded IL-13 expression and in turn attenuated Th2-cytokines, BLT2, NF-κB, and IgE levels to ameliorate AHR. Thus, our findings suggested that morin could be considered as a potential alternative therapeutic agent for the management of allergic asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Flavonoids; Hemodynamics; Immunoglobulin E; Interleukin-13; Lung; Male; NF-kappa B; Ovalbumin; Oxygen; Rats; Rats, Sprague-Dawley; Receptors, Leukotriene B4; Signal Transduction; Sulfatases; Superoxide Dismutase

Topics: Actins; Airway Remodeling; Animals; Antigens; Asthma; beta Catenin; Collagen; Down-Regulation; Lung; Male; Ovalbumin; Rats, Sprague-Dawley; Vitamin D; Vitamins; Wnt Signaling Pathway; Wnt-5a Protein

S-Allyl cysteine (SAC) is an active component in garlic and has various pharmacological effects, such as anti-inflammatory, anti-oxidant, and anti-cancer activities. In this study, we explored the suppressive effects of SAC on allergic airway inflammation induced in an ovalbumin (OVA)-induced asthma mouse model. To induce asthma, BALB/c mice were sensitized to OVA on days 0 and 14 by intraperitoneal injection and exposed to OVA from days 21 to 23 using a nebulizer. SAC was administered to mice by oral gavage at a dose of 10 or 20 mg/kg from days 18 to 23. SAC significantly reduced airway hyperresponsiveness, inflammatory cell counts, and Th2 type cytokines in bronchoalveolar lavage fluid induced by OVA exposure, which was accompanied by reduced serum OVA-specific immunoglobulin E. In histological analysis of the lung tissue, administration of SAC reduced inflammatory cell accumulation into lung tissue and mucus production in airway goblet cells induced by OVA exposure. Additionally, SAC significantly decreased MUC5AC expression and nuclear factor-κB phosphorylation induced by OVA exposure. In summary, SAC effectively suppressed allergic airway inflammation and mucus production in OVA-challenged asthmatic mice. Therefore, SAC shows potential for use in treating allergic asthma.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cysteine; Cytokines; Disease Models, Animal; Eosinophilia; Female; Immunoglobulin E; Lung; Mice, Inbred BALB C; Mucus; Ovalbumin

Exposure to early-life undernutrition is closely related to higher risks of adverse immunologic outcomes in adulthood. Although it has been suggested that asthma has its origins in early life, its underlying mechanisms remain largely unknown.. We characterized the effects of early-life undernutrition on T lymphocytes, which play a pivotal role in immune diseases, and we investigated whether this contributes to susceptibility to asthma in adulthood.. Pregnant mice were fed a protein restriction diet (PRD) to establish an early-life undernutrition model. Naive CD4. PRD CD4. Early-life undernutrition induced mechanistic target of rapamycin 1-dependent glycolysis upregulation and T

Topics: Allergens; Animals; Asthma; CD4-Positive T-Lymphocytes; Cytokines; Epigenesis, Genetic; Female; Glycolysis; Lung; Male; Malnutrition; Mechanistic Target of Rapamycin Complex 1; Mechanistic Target of Rapamycin Complex 2; Mice, Inbred C57BL; Ovalbumin; Pregnancy

Cyanidin-3-O-β-glucoside (Cy-3-g), a typical and abundant monomer of anthocyanins, exhibits a variety of biological activities, such as anti-atherosclerosis, anti-obesity, and anticancer effects. However, to date little is known about its effects on asthma. This study aimed to investigate the efficacy of dietary Cy-3-g on allergic asthma in an animal model. BALB/c mice were sensitized and challenged with ovalbumin (OVA) to induce allergic asthma. The pathological changes of the lung tissues, type 2 helper (Th2)-associated cytokine production in bronchoalveolar lavage fluid (BALF), and the interleukin 4 receptor alpha (IL-4Rα)-signal transducer and activator of transcription 6 (STAT6) signaling pathway activities were assessed. We found that Cy-3-g significantly inhibited OVA-induced inflammatory cell infiltration and mucus hyper-production in lung tissues, reduced the production of interleukin 4 (IL-4), interleukin 5 (IL-5) and interleukin 13 (IL-13) in BALF. Furthermore, Cy-3-g effectively suppressed OVA-induced up-regulation of the IL-4Rα-STAT6 signaling pathway activity of the lung tissues. These results demonstrated that dietary Cy-3-g could attenuate allergic airway inflammation in a murine asthma model, and Cy-3-g might be used as an agent for asthma prevention and/or treatment in the future.

Topics: Allergens; Animals; Anthocyanins; Anti-Inflammatory Agents; Asthma; Cytokines; Dietary Supplements; Disease Models, Animal; Female; Glucosides; Humans; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Cell Surface; Respiratory Hypersensitivity; STAT6 Transcription Factor; Th2 Cells

Identification of the characterization of cysteinyl leukotrienes receptor (CysLTRs) could facilitate our understanding of these receptors' role in asthma. We aimed to investigate the localization and interactions of CysLTRs using a mouse model of asthma. BALB/c mice were administered ovalbumin (OVA) to induce allergic asthma. Some mice were administered the antagonists of CysLTR1, CysLTR2, and purinergic receptor P2Y12 (P2Y12R) (montelukast, HAMI 3379 and clopidogrel, respectively). The expression levels of CysLTR1, CysLTR2, and P2Y12R on lung tissues and inflammatory cells were evaluated by western blot, flow cytometry, and immunochemistry. CysLTR1 and P2Y12R were significantly up-regulated in lung tissues (P < 0.05 for each) from mouse after being sensitized and challenged with OVA (OVA/OVA). The ratio of CysLTR1: CysLTR2: P2Y12R in lungs of negative control (NC) mice was shifted from 1:0.43:0.35 to 1:0.65:1.34 in OVA/OVA mice. Montelukast significantly diminished the up-regulation of CysLTR1, CysLTR2, and P2Y12R (P < 0.05 for each), while the effects of HAMI 3379 and clopidogrel were predominant on the expression of CysLTR2 and P2Y12R, respectively. Montelukast predominantly diminished the cell count, while clopidogrel potently inhibited the release of interleukin (IL)-4, IL-5, and IL-13. Our study demonstrated the interactions between CysLTRs, thereby highlighting the potential synergistic effects of CysLTR antagonists in asthma treatment.

Topics: Acetates; Animals; Asthma; Clopidogrel; Cyclohexanecarboxylic Acids; Cyclopropanes; Disease Models, Animal; Drug Therapy, Combination; Eosinophils; Female; Inflammation; Interleukins; Leukotriene Antagonists; Mice; Mice, Inbred BALB C; Ovalbumin; Phthalic Acids; Purinergic P2Y Receptor Antagonists; Quinolines; Receptors, Leukotriene; Receptors, Purinergic P2Y12; Sulfides; Th2 Cells

Clinical evidence gathered in Chinese communities suggested that acupoint sticking therapy could be an alternative treatment for asthma-related diseases. However, its underlying mechanism is still poorly understood.. In this study, we aimed to investigate the mechanism of the anti-inflammatory effect of acupoint sticking application with 'Treatment of Winter Disease in Summer' (TWDS) prescription by using metabolomics.. Allergic asthma in guinea pig was sensitized and challenged by ovalbumin (OVA). Histopathological evaluation of the lung tissue was performed by hematoxylin and eosin (H&E) staining and Masson's trichrome staining. The levels of Th2 cytokine and IgE level in serum were measured using enzyme-linked immunoassay (ELISA). The mRNA expression levels of IL-4, IL-5, IL-13 and orosomucoid-like 3 (ORMDL3) were measured using quantitative reverse transcription polymerase chain reaction (RT-qPCR). Proteins of NF-κB signaling pathway were measured using western blot. The serum metabolomics profiles were obtained by using ultra-performance liquid chromatography combined with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS).. The overall results confirmed that AST with TWDS prescription had a significant protective effect against OVA-induced allergic asthma in guinea pig. This treatment not only attenuated airway inflammation and collagen deposition in the airway, but also decreased the levels of IL-4, IL-5, IL-13 and IgE in serum. In addition, metabolomics results indicated that metabolisms of phospholipid, sphingolipid, purine, amino acid and level of epinephrine were restored back to the normal control level. Moreover, results of the gene expression of ORMDL3 in lung tissues indicated that AST using TWDS could alter the sphingolipid metabolism. Further western blotting analysis also showed that its anti-inflammatory mechanism was by decreasing the phosphorylation of p65 and IκB.. The study demonstrated that metabolomics provides a better understanding of the actions of TWDS acupoint sticking therapy on OVA-induced allergic asthma.

Topics: Acupuncture Therapy; Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Drugs, Chinese Herbal; Guinea Pigs; Hypersensitivity; Immunoglobulin E; Lung; Male; Membrane Proteins; Metabolomics; NF-kappa B; Ovalbumin; Signal Transduction

Reproductive organs play a pivotal role in asthma development and progression, especially in women. Endocrine environment changes associated with the menstrual cycle, pregnancy, and menopause can exacerbate the clinical features of asthma. Factors secreted by reproductive organs may be responsible for the gender difference and age-related changes in adult asthma. Here, we show that mammalian seminal fluid has anti-asthma effects exclusively in females. Exposure to murine seminal fluid markedly reduced eosinophilic airway inflammation in 2-month-old female mice upon ovalbumin inhalation. The anti-asthma effect with seminal fluid from 10-month-old males was double that with fluid from 2-month-old males, suggesting that it depended on male sexual maturation. We further found that seminal fluid from middle-aged human volunteers had beneficial effects in asthmatic female mice; these effects were associated with transcriptional repression of osteopontin and IL-17A, which are poor prognostic factors for asthma. In 2-month-old male mice, however, human seminal fluid failed to decrease asthmatic features and even enhanced osteopontin and IL-17A transcription. Our data demonstrate that age-related seminal fluid exerts opposing effects in asthmatic male and female mice. These findings may help the development of novel approaches to control the prevalence and age-related progression of asthma in women.

Topics: Adult; Aging; Animals; Asthma; Bone Marrow Cells; Bronchoalveolar Lavage Fluid; Dendritic Cells; Female; Humans; Immunoglobulin E; Interleukin-13; Male; Mice; Middle Aged; Ovalbumin; Semen; Sex Characteristics

Asthma is characterized by chronic airway inflammation, which is the underlying cause of airway remodeling featured by goblet cell hyperplasia, subepithelial fibrosis, and proliferation of smooth muscle. Sevoflurane has been used to treat life-threatening asthma and our previous study shows that sevoflurane inhibits acute lung inflammation in ovalbumin (OVA)-induced allergic mice. However, the effect of sevoflurane on airway remodeling in the context of chronic airway inflammation and the underlying mechanism are still unknown. Here, female C57BL/6 mice were used to establish chronic airway inflammation model. Hematoxylin and eosin (H&E), periodic acid-Schiff (PAS), and Sirius red (SR) staining were used to evaluate airway remodeling. Protein levels of α-SMA, VEGF, and TGF-β1 in lung tissues were detected by western blotting analyses and immunohistochemistry staining. Results showed that inhalation of sevoflurane inhibited chronic airway inflammation including inflammatory cell infiltration and pro-inflammatory cytokine production in BALF of the OVA-challenged mice. Meanwhile, sevoflurane suppressed airway thickening, goblet cell hyperplasia, smooth muscle hyperplasia, collagen deposition, and fiber hyperplasia in the lung tissues of the mice with airway remodeling. Most notably, sevoflurane inhibited the OVA-induced expressions of VEGF and TGF-β1. These results suggested that sevoflurane effectively inhibits airway remodeling in mouse model of chronic airway inflammation, which may be due to the downregulation of VEGF and TGF-β1in lung tissues. Therefore, our results indicate a potential role of sevoflurane in inhibiting airway remodeling besides its known suppression effect on airway inflammation, and support the use of sevoflurane in treating severe asthma in ICU.

Topics: Airway Remodeling; Anesthetics, Inhalation; Animals; Asthma; Down-Regulation; Female; Inflammation; Mice; Ovalbumin; Sevoflurane; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

There has been considerable research on the involvement of RhoA/Rho kinase signalling in smooth muscle contractions. However, only a few reports have addressed the specific role of Rac1, which is a member of the Rho GTPase superfamily. Therefore, this study investigated the role of Rac1-related pathways in bronchial smooth muscle (BSM) contractions. Bronchial rings isolated from mice were suspended in an organ bath, and the isometric contractions of circular smooth muscles were monitored. The phosphorylation of myosin light chains (MLCs) was analysed by immunoblotting. The Rac1 inhibitor EHT1864 inhibited carbachol (CCh)-induced BSM contractions, although high K

Topics: Animals; Asthma; Bronchi; Carbachol; Disease Models, Animal; Humans; Male; Mice; Muscle Contraction; Muscle, Smooth; Neuropeptides; Ovalbumin; Pyrones; Quinolines; rac1 GTP-Binding Protein; Signal Transduction; Up-Regulation

Allergic asthma causes morbidity in many subjects, and novel precision-directed treatments would be valuable.. We sought to examine the role of a novel innate molecule, repulsive guidance molecule b (RGMb), in murine models of allergic asthma.. In models of allergic asthma using ovalbumin or cockroach allergen, mice were treated with anti-RGMb or control mAb and examined for airway inflammation and airway hyperreactivity (AHR), a cardinal feature of asthma. The mechanisms by which RGMb causes airways disease were also examined.. We found that blockade of RGMb by treatment with anti-RGMb mAb effectively blocked the development of airway inflammation and AHR. Importantly, blockade of RGMb completely blocked the development of airway inflammation and AHR, even if treatment occurred only during the challenge (effector) phase. IL-25 played an important role in these models of asthma because IL-25 receptor-deficient mice did not develop disease after sensitization and challenge with allergen. RGMb was expressed primarily by innate cells in the lungs, including bronchial epithelial cells (known producers of IL-25), activated eosinophils, and interstitial macrophages, which in the inflamed lung expressed the IL-25 receptor and produced IL-5 and IL-13. We also found that neogenin, the canonical receptor for RGMb, was expressed by interstitial macrophages and bronchial epithelial cells in the inflamed lung, suggesting that an innate RGMb-neogenin axis might modulate allergic asthma.. These results demonstrate an important role for a novel innate pathway in regulating type 2 inflammation in patients with allergic asthma involving RGMb and RGMb-expressing cells, such as interstitial macrophages and bronchial epithelial cells. Moreover, targeting this previously unappreciated innate pathway might provide an important treatment option for allergic asthma.

Topics: Allergens; Animals; Antibodies, Monoclonal; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cell Adhesion Molecules, Neuronal; Cockroaches; Female; Interleukin-1 Receptor-Like 1 Protein; Macrophages; Membrane Proteins; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Programmed Cell Death 1 Ligand 2 Protein; Receptors, Interleukin

Sphingosine-1-phosphate 2 (S1P. Using an ovalbumin (OVA)-induced asthma model, the function of S1P. Eosinophil accumulation and elevated Th2 cytokine levels in bronchoalveolar lavage fluid and inflamed lung tissues were strongly inhibited by administration of JTE-013 before OVA sensitization, before OVA challenge, and before both events. In S1P. Pro-allergic functions of S1P

Topics: Adoptive Transfer; Allergens; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dendritic Cells; Eosinophils; Female; Lung; Mast Cells; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Pyrazoles; Pyridines; Sphingosine-1-Phosphate Receptors

Myristoylated alanine-rich C kinase substrate (MARCKS), a PKC substrate, facilitates mucus production and neutrophil migration. However, the effects of therapeutic procedures targeting the phosphorylation site of MARCKS on steroid-resistant asthma and the mechanisms underlying such effects have not yet been investigated. We designed a peptide that targets the MARCKS phosphorylation site (MPS peptide) and assessed its therapeutic potential against steroid-resistant asthma.. Mice were sensitized with ovalbumin (OVA), alum, and challenged with aerosolized OVA five times a week for 1 month. The mice were intratracheally administered MPS peptides three times a week, 1 hr before OVA challenge. Asthma symptoms and cell profiles in the bronchoalveolar lavage were assessed, and key proteins were analysed using Western blotting.. Phosphorylated (p)-MARCKS was highly expressed in inflammatory and bronchial epithelial cells in OVA-immunized mice. MPS peptide reduced eosinophils, neutrophils, mucus production, collagen deposition, and airway hyper-responsiveness. Dexamethasone (Dexa) did not alleviate steroid-resistant asthma symptoms. MPS peptide caused a decrease in p-MARCKS, nitrotyrosine and the expression of oxidative stress enzymes, NADPH oxidase dual oxidase 1 and inducible NOS, in lung tissues. Compared to Dexa, MPS peptides inhibited C5a production and attenuated IL-17A and KC production in the airway more effectively, thus suppressing asthma symptoms.. Our findings indicate that targeting MARCKS phosphorylation through MPS treatment may inhibit neutrophilic inflammation and relieve asthma symptoms, thereby highlighting its potential as a therapeutic agent for steroid-resistant asthma.

Topics: Adrenal Cortex Hormones; Allergens; Animals; Asthma; Disease Models, Animal; Drug Resistance; Epithelial Cells; Female; Lung; Mice, Inbred BALB C; Myristoylated Alanine-Rich C Kinase Substrate; Ovalbumin; Peptides; Phosphorylation

Various studies have shown that flavones have several pharmacological activities including anti-allergy activities. However, the bioavailability of oral flavones is very low, and whether inhaled administration can improve efficacy in respiratory disease models is unclear. In the present study, the anti-allergic activities of inhaling 5,7-dimethoxy-3,4'-dihydroxyflavone (MHF), a synthetic flavonoid, was investigated by comparison with disodium cromoglycate (DSCG) and nedocromil sodium (NS) in rat allergic models. In an anti-DNP-IgE-induced asthmatic model, inhaled MHF dose-dependently inhibited the increase in airway resistance after antigen challenge. In an ovalbumin (OVA)-induced asthmatic model, inhaled MHF showed significant suppression of airway hyperresponsiveness; a decrease in eosinophil and neutrophil counts, IL-4, IL-5 and leukotriene D

Topics: Administration, Inhalation; Animals; Anti-Allergic Agents; Asthma; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Flavonoids; Ovalbumin; Rats; Rats, Sprague-Dawley

Around the globe, worsening air pollution is spawning major public health and environmental concerns, especially in the poorest and most populous cities. As a major secondary air pollutant, ozone is a potential risk factor for exacerbated asthma, although the underlying mechanisms remain uncertain. In this study, we aim to investigate the role of ozone on asthma exacerbation using a classic asthmatic model with allergic airway inflammation by treating Balb/c mice with ovalbumin (OVA). Our study shows ozone exposure significantly exacerbated OVA-induced asthmatic phenotypes, including serum immunoglobulin, Th cytokines, inflammatory cell counts, mucus production, airway remodeling, and airway hyper-responsiveness (AHR). Interestingly, expression of transient receptor potential cation channel subfamily V member1 (TRPV1) was also significantly elevated in ozone-exacerbated asthmatic mice and that treatment with TRPV1 antagonist effectively suppressed AHR, airway inflammation and remodeling. The underlying mechanisms of these effects may be associated with suppression of neuropeptide calcitonin gene-related peptide (CGRP) and thymic stromal lymphopoietin (TSLP), an epithelial cell-derived cytokine. Base on the role of TRPV1 in allergic asthma, this study further revealed that inhibition of TRPV1 by TRPV1 antagonist has significant anti-inflammatory effects on ozone-induced asthma exacerbation in this study. Induction of TRPV1 expression may be an important mechanism underlying the increased risks for asthma after exposure to environmental pollutants.

Topics: Air Pollutants; Animals; Asthma; Cytokines; Disease Models, Animal; Inflammation; Mice; Mice, Inbred BALB C; Ovalbumin; Ozone; Respiratory System; Thymic Stromal Lymphopoietin; Transient Receptor Potential Channels; TRPV Cation Channels

The activation of peroxisome proliferator-activated receptor γ (PPAR-γ) has been shown to attenuate allergic airway inflammation (AAI). To gain better understanding of mechanisms underlying this effect, the impact of rosiglitazone (RSG), a PPAR-γ agonist, on CD4

Topics: Animals; Asthma; CD4 Lymphocyte Count; Disease Models, Animal; Forkhead Transcription Factors; Gene Expression; Hypersensitivity; Mice; Mice, Inbred BALB C; Ovalbumin; PPAR gamma; Rosiglitazone; T-Lymphocytes, Regulatory; Treatment Outcome

With the increasing morbidity and mortality of asthma, asthma aggravated by environmental pollution has drawn more attention. This study investigated the exacerbating effects of trimellitic anhydride (TMA), a typical pollutant, in ovalbumin (OVA)-induced asthmatic mice and the gene and protein expressions of TRPA1, V1, V2 in lung tissue. Female BALB/c mice were respectively administered for 42 days as follow: sensitized and challenged with OVA, sensitized and challenged with TMA, sensitized with OVA and challenged with OVA plus TMA, as well as sensitized and challenged with OVA plus TMA. 24 h after the last challenge, the changes in airway resistance (RI) and lung dynamic compliance (Cdyn) were tested. The levels of the inflammatory cells in blood and bronchoalveolar lavage fluid (BALF) were determined. The gene and protein expressions of TRPA1, V1, V2 in lung tissue were examined, and levels of interleukin (IL)-4, -13, substance P (SP), prostaglandin D

Topics: Allergens; Animals; Asthma; Calcium Channels; Disease Models, Animal; Environmental Pollutants; Female; Gene Expression Regulation; Humans; Lung; Mice; Ovalbumin; Phthalic Anhydrides; TRPA1 Cation Channel; TRPV Cation Channels

Unresolved inflammation underpins the pathogenesis of allergic airway diseases, such as asthma. Ketamine, accepted as a promising therapy for resistant asthma, has been demonstrated to attenuate allergic airway inflammation. However, the anti-inflammatory mechanism by ketamine in this setting is largely unknown. We aimed to investigate whether autophagy was involved in the protective effect of ketamine on allergic airway inflammation. Female C57BL/6 mice were sensitized to ovalbumin (OVA) and treated with ketamine at 25, 50, or 100 mg/kg prior to OVA challenge. In this model, the pulmonary morphological findings and airway inflammation were significantly inhibited at 50 mg/kg but not at 25 or 100 mg/kg. Moreover, 50 mg/kg ketamine abrogated the increased concentrations of inflammatory cytokines in bronchoalveolar lavage fluid (BALF) of allergic mice, as well as activated the expression of phosphorylated mammalian target of rapamycin (p-MTOR) and inhibited autophagy in allergic mice. To confirm whether the effect of 50 mg/kg ketamine on asthma was mediated by inhibiting autophagy, rapamycin was administered to mice sensitized to OVA and exposed to 50 mg/kg ketamine. All of the effect of 50 mg/kg ketamine was reversed by rapamycin treatment, including increased p-MTOR and decreased autophagy. Taken together, the present study demonstrates that 50 mg/kg ketamine inhibits allergic airway inflammation by suppressed autophagy, and this effect is mediated by the activation of MTOR in the lungs of allergic mice.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Autophagy; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Inflammation; Ketamine; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; TOR Serine-Threonine Kinases

Corticosteroids are the preferred option to treat asthma, however, they possess serious side effects and are inefficient in 10% of patients. Thus, new therapeutic approaches for asthma treatment are required.. To study the efficacy of a novel glutarimide derivative XC8 in a Sephadex-induced lung inflammation in rats as well as in acute and chronic ovalbumin-induced allergic asthma in guinea pigs.. Rats were treated with 0.18-18 mg/kg of XC8 intragastrically 4 times (24 h and 1 h prior to and 24 h and 45 h after endotracheal administration of Sephadex). The number of inflammatory cells in bronchoalveaolar lavages (BAL) was determined. Guinea pigs were treated with 0.045 -1.4 mg/kg (acute asthma) or with 1.4 and 7.0 mg/kg of XC8 (chronic asthma) intragastrically following the sensitization with ovalbumin and during aerosol challenge. Lung inflammation, numbers of eosinophils (BAL and lung tissue), goblet cells, degranulating mast cells and specific airway resistance (sRAW) were determined. The comparator steroid drug budesonide (0.5 mg/kg for rats and 0.16 mg/kg for guinea pigs) was administered by inhalation.. XC8 reduced influx of eosinophils into BAL in Sephadex-induced lung inflammation model in rats (by 2.6-6.4 times). Treatment of acute asthma in guinea pigs significantly reduced eosinophils in guinea pigs in BAL (from 55% to 30%-39% of the total cell count) and goblet cells in lung tissue. In a model of acute and chronic asthma, XC8 reduced significantly the number of eosinophils and degranulating mast cells in the lung tissue. Treatment with XC8 but not with budesonide decreased the specific airway resistance in acute and chronic asthma model up to the level of naive animals.. XC8 induced a profound anti-inflammatory effect by reducing eosinophils in BAL and eosinophils and degranulating mast cell numbers in the airway tissue. The anti-asthmatic effect of XC8 is comparable to that of budesonide. Moreover, in contrast to budesonide, XC8 was capable to reduce goblet cells and airway resistance.

Topics: Administration, Oral; Animals; Asthma; Budesonide; Dextrans; Eosinophils; Guinea Pigs; Male; Ovalbumin; Piperidones; Pneumonia; Rats; Rats, Wistar

Asthma is a chronic lung disease characterized by an imbalance of T‑helper (Th)1/Th2 cells and their cytokine profiles. Natural killer (NK) cells constitute a considerable subset of the lymphocyte population in the lungs, and provide protection against respiratory infection by fungi, bacteria and viruses. However, the mechanism by which NK cells are involved in asthma remains to be fully elucidated. The present study analyzed the dynamic changes of NK cells and their subsets during the development of the ovalbumin (OVA)‑induced allergic airway response. Lung tissues were histologically examined for cell infiltration and mucus hypersecretion. The number, activity and cytokine‑secreting ability of NK cells was determined by flow cytometry. The results showed that the percentage of NK cells in the lung was decreased following OVA sensitization and challenge. However, NK cells exhibited enhanced activity and secreted more Th2 cytokines (IL‑5 and IL‑13) following OVA challenge. Furthermore, the proportion of CD11b‑ NK subsets increased with the development of asthma, and CD11b‑ CD27‑ NK cells were the primary NK subset producing Th2 cytokines. These findings suggest that, although NK cells are not the crucial type of lymphocytes involved in asthma, OVA induces NK cells to secrete Th2 cytokines that may be involved in the pathogenesis of asthma.

Topics: Allergens; Animals; Asthma; Biomarkers; Disease Models, Animal; Female; Immunophenotyping; Interleukin-13; Interleukin-5; Killer Cells, Natural; Mice; Ovalbumin; Th2 Cells

BACKGROUND Bulleyaconitine A (BLA) has been widely used as analgesic against chronic inflammatory pain in China. However, its potential therapeutic role in asthma remains unclear. The purpose of this study was to investigate the effect of BLA on airway inflammation in mice with allergic asthma. MATERIAL AND METHODS Specific-pathogen-free (SPF) female Balb/c mice were randomly divided into the following 6 groups: (1) Control group (NC), (2) Asthma group (AS), (3) BLA-L group, (4) BLA-M group, (5) BLA-H group, and (6) Dexamethasone group. An asthma mouse model was established by administration of ovalbumin (OVA) and mice were sacrificed within 24 h after the last challenge. Enzyme-linked immunosorbent assay (ELISA) method was used to determine the relative expression levels of IgE and IgG in mouse serum. In addition, bronchoalveolar lavage fluid (BALF) was collected and IL-4, TNF-α, and MCP-1 levels were determined by ELISA. Furthermore, eosinophils, lymphocytes, and macrophages in BALF were classified and analyzed, and inflammatory cell infiltration in the airways of mice was determined by hematoxylin-eosin (HE) staining. The expression of NF-κB1 and PKC-δ in mouse lung tissue was determined by Western blot analysis. RESULTS The levels of serum IgE and IgG in BLA- or Dex- treated mice were significantly reduced compared to those in the asthma (AS) group (P<0.01), whereas the levels of cytokines IL-4, TNF-α, and MCP-1 were significantly decreased (P<0.01). HE-staining showed that BLA significantly reduced inflammatory cell infiltration and mucus secretion in lung tissue. Moreover, BLA inhibited the expression of NF-κB1 and PKC-d via the NF-κB signaling pathway in the lung. CONCLUSIONS Our data show that BLA activates PKC-δ/NF-κB to reduce airway inflammation in allergic asthma mice.

Topics: Aconitine; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Chemokine CCL2; China; Cytokines; Disease Models, Animal; Eosinophils; Female; Inflammation; Interleukin-4; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Protein Kinase C-delta; Tumor Necrosis Factor-alpha

San'ao decoction (SAD), a traditional Chinese prescription, is well-known in asthma treatment. In the current study, the protective role of SAD and its mechanism in aggravated asthma mice model via regulation of TRP channel were evaluated and explored.. UPLC-QTOF-MS was used for analyzing the chemicals in SAD. The major chemical components in SAD were separated and detected under an optimized chromatographic and MS condition. 75 BALB/c mice were randomly divided into five groups: normal group, model group, dexamethasone group (0.75 mg kg. 21 signal peaks of the chemicals in SAD were identified with the method of UPLC-QTOF-MS. SAD, especially SAD-high dose exerted significant effects on OVA plus PM2.5 mice model in relieving lung injury score (P < 0.05), reducing eosinophil (EOS) count in blood (P < 0.05) and inflammatory cells ratio in BALF (P < 0.05, P < 0.01), decreasing RI value (P < 0.05) while increasing Cdyn value (P < 0.05), reducing IL-13, PGD. SAD could improve pulmonary functions, relieve lung injury, as well as reduce IL-13, PGD

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Lung; Lung Injury; Mice; Mice, Inbred BALB C; Ovalbumin; Particulate Matter; Respiratory Function Tests; TRPA1 Cation Channel; TRPV Cation Channels

Epidemiological and clinical studies have demonstrated a close association between obesity and asthma. The current study investigated the effect of high-fat diet on tracheal responsiveness to methacholine and insulin resistance in ovalbumin (OVA) sensitized male and female rats. The rats were divided into eight groups (n=6 per group): female with the normal diet (F+ND), male with the normal diet (M+ND), female OVA-sensitized with the normal diet (F+SND), male OVA-sensitized with the normal diet (M+SND), female with high-fat diet (F+HFD), male with high-fat diet (M+HFD), female OVA-sensitized with high-fat diet (F+SHFD), and male OVA-sensitized with high-fat diet (M+SHFD). All rats were fed for 8 weeks with high-fat diet or standard pelts, and for another 4 weeks, they were sensitized with OVA or saline. At the end of the study, the tracheal responsiveness to methacholine, serum insulin, and blood glucose levels was measured. Also, insulin resistance indexes were determined. OVA-sensitization and diet-induced obesity caused the curve of methacholine concentration response to shifting to the left. In addition, results indicated that the EC50 (the effective concentration of methacholine generating 50% of peak response) in F+SHFD rats was statistically lower than M+SHFD group (p<0.05). Moreover, insulin resistance was higher in the F+SHFD than the M+SHFD group (p<0.001). These results suggest that insulin resistance and metabolic syndrome may be involved in the pathogenesis of obesity associated with OVA-sensitized rats condition, especially in female animals.

Topics: Allergens; Animals; Asthma; Blood Glucose; Bronchoconstrictor Agents; Diet, High-Fat; Disease Models, Animal; Female; Insulin; Insulin Resistance; Male; Methacholine Chloride; Obesity; Ovalbumin; Rats, Wistar; Trachea

The role of IL-38, a new member of the IL-1 family, in airway eosinophilic inflammatory conditions such as asthma is unclear. To investigate the role of IL-38 in airway eosinophilic inflammation, an IL-38-gene deficient (KO) murine asthma model was analyzed.. The numbers of eosinophils and neutrophils, and levels of IL-5, IL-13 and IL-17A protein and mRNA in bronchoalveolar lavage fluid (BALF) and lung tissue were compared between wild-type (WT) and IL-38-KO mice after OVA sensitization and challenge. The effects of additional purified recombinant mouse (rm) IL-38 protein were investigated in the IL-38-KO murine asthma model.. The IL-38 and IL-5 mRNA in WT mice was significantly higher after OVA challenge than after saline challenge (p<0.05). The number of airway eosinophils in IL-38-KO mice was significantly lower than in WT mice after OVA challenge (p<0.01). BALF analysis confirmed the lower number of airway eosinophils in IL-38-KO mice and showed that this was significantly associated with lower IL-5 protein levels (r=0.92, p<0.0001). However, the additional rm IL-38 protein did not neutralize airway eosinophilia in IL-38-KO mice.. IL-38 may enhance airway eosinophilic inflammation in asthma through IL-5 induction.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Crosses, Genetic; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Eosinophilia; Female; Inflammation; Interleukin-1; Interleukin-13; Interleukin-17; Interleukin-5; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophils; Ovalbumin; Recombinant Proteins; RNA, Messenger

The possibility has been suggested that interferon (IFN)-λs can be induced rapidly for restricting respiratory viral infection in asthmatic mice and may modulate Th2-related immune responses that underlie the pathogenesis of asthma. We sought to determine the in vivo contribution of IFN-λs on decrease of Th2 cytokines in the respiratory tract of in vivo asthma. Lungs of asthmatic mice were severely inflamed, with extensive inflammatory cell infiltration and increased goblet cell metaplasia with higher total lung resistance. The mean protein levels of TSLP and IL-33 from BAL fluid of asthmatic mice were significantly higher until 7 days. Following the collection of lung tissue of 20 asthmatic mice, TSLP and IL-33 gene expressions inversely correlated with mRNA levels of IFN-λ

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Epithelial Cells; Inflammation; Interleukin-33; Mice; Mice, Inbred C57BL; Ovalbumin; Th2 Cells

Mouse models of allergic asthma play a crucial role in exploring of asthma pathogenesis and testing of novel anti-inflammatory drugs. Widely used acute asthma models usually developed with adjuvant (aluminum hydroxide (alum)) do not reproduce one of the main asthma feature - airway remodeling while chronic asthma model mimic the pathophysiology of human disease. Moreover, the use of alum causes distress in experimental animals and impedes the test of adjuvant-containing drugs. In this study, we aimed to develop a chronic adjuvant-free asthma model with pronounced asthmatic phenotype.. Female BALB/c mice were divided into 3 groups. The first group was sensitized with intraperitoneal injections of ovalbumin (OVA) emulsified in aluminum hydroxide on days 0, 14, 28 followed by two stages of intranasally challenge with OVA on days 41-43 and 62-64. The second group was subcutaneously sensitized with the same dose of OVA without adjuvant and challenged on the same days. The third group (negative control) included mice which did not received any kind of treatment (i.e. sensitization and challenge). Serum levels of OVA-specific IgE, IgG2a and IgG1 antibodies were detected by ELISA. Airway hyper-responsiveness was measured by non-invasive plethysmography on days 44 and 65. Bronchoalveolar lavage fluids (BALF) sampled in all groups on days 45 and 66 were analyzed by light microscopy. The left lung was removed for histological analysis. The IL-4 and IFNγ mRNA expression in BALF cells was evaluated by RT-PCR.. The OVA-specific IgE antibody response was two-fold increased in mice from adjuvant-free group compared to the adjuvant group that reflects reorientation of immune response towards Th2 phenotype. At the same time, the level of OVA-specific IgG1 and IgG2a antibodies was increased in the adjuvant group. Airway hyperresponsiveness to methacholine in mice of both experimental groups was two-fold higher than in control. Analysis of cell composition in BAL has shown a significant increase in eosinophil count in both experimental groups that indicate the development of allergic inflammation. Lung histology revealed airway remodeling in both experimental groups including goblet cell hyperplasia/metaplasia, thickening of airway walls, collagen deposition in the wall of distal airways. Additionally, the tendency to develop hypertrophy of bronchial smooth muscle layer was observed. Study of gene expression in BAL cells revealed the increase of IL-4 level in both adjuvant and adjuvant-free groups while IFNγ expression in both experimental groups was similar to control group.. We have developed a chronic adjuvant-free mouse asthma model which possesses all necessary features of the disease including airway remodeling and is more suitable for pre-clinical evaluation of novel therapeutic approaches including adjuvant-containing drugs.

Topics: Adjuvants, Immunologic; Airway Remodeling; Aluminum Hydroxide; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Disease Progression; Female; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-4; Lung; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; Th2 Cells; Time Factors

Exposure to ozone contributed to the worsening of inflammation and glucocorticoids insensitivity in OVA-challenged asthma. Interleukin-17A participates centrally in stages of the inflammatory response and glucocorticoids insensitivity. In this study, the effect of neutralizing anti-murine interleukin-17A monoclonal antibody (IL-17A mAb) on inflammation and glucocorticoids insensitivity in ozone-exposed and ovalbumin (OVA)-challenged mice was investigated.. Mice were sensitized and challenged with OVA and then exposed to ozone. Dexamethasone (Dex) and IL-17A mAb were administrated in corresponding periods.. Compared with OVA-challenged mice, combination administration of ozone exposure and OVA challenge increased the recruitment of inflammatory cells in bronchoalveolar lavage fluid, enhanced the inflammation scores and levels of inflammatory cytokines and IL-17A mRNA, and caused the activation of p38 MAPK together with down regulation of glucocorticoids recepters (GR) in lung tissue. Monotherapy of IL-17A mAb partially attenuated lung inflammation in OVA-challenged and ozone-exposed mice, while the combination treatment of Dex and IL-17A mAb effectively reduced lung inflammation, inactivated p38 MAPK and up regulated GR in lung tissue.. Ozone exposure worsened OVA-challenged airway inflammation, activation of p38 MAPK and down regulation of GR in OVA-sensitized and -challenged mice, which was effectively counteracted by IL-17A mAb, and combination treatment of IL-17A mAb and Dex shows profound efficacy in inhibiting airway inflammation and improving glucocorticoids insensitivity synergistically.

Topics: Animals; Antibodies, Monoclonal; Antibodies, Neutralizing; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dexamethasone; Disease Models, Animal; Glucocorticoids; Interleukin-17; Lung; Male; Mice; Ovalbumin; Ozone; p38 Mitogen-Activated Protein Kinases; Pneumonia

Spatial and temporal control of gene expression using cre/loxP technology is a major methodological advance for biomedical research. The ability to alter gene expression after an in vivo disease model has been established and allows researchers the opportunity to examine the impact of that gene on the perpetuation of a disease, a mechanistic insight that is arguably more therapeutically relevant than developmental mechanisms.We used the cre/LoxP technology in mice to show that β-arrestin-2, a gene previously shown to be required for the development of the asthma phenotype, is also required for the perpetuation of, at least, the airway hyperresponsiveness characteristic of that phenotype. Here we describe stepwise methods for the activation of the cre-loxP technology and induction of murine allergic inflammatory airway disease. We comment on the unanticipated problems encountered, which we speculate were a result of interactions between the allergen and β-arrestin-2 gene (Arrb2) regulation and the effect of tamoxifen on the asthma phenotype. The issues encountered here may be generally applicable to cre/loxP utilization in inflammatory disease models, especially if the gene of interest is associated with the inflammatory cascade.

Topics: Animals; Asthma; beta-Arrestin 2; Disease Models, Animal; Hypersensitivity; Inflammation; Mice, Knockout; Molecular Biology; Ovalbumin

Despite the reported role of poly(ADP-ribose) polymerase (PARP) in asthma inflammation, its contribution during remodeling is not clearly known. The main aim of the current investigation was to examine the potential of olaparib, a pharmacological inhibitor of PARP against airway remodeling using an ovalbumin (OVA)-based murine model of chronic asthma. The results demonstrated that post-challenge olaparib treatment (5 mg/kg i.p., 30 min after OVA exposure) for six weeks (3 days/week) attenuates inflammation, mucus production, and collagen deposition in lungs. Additionally, olaparib blunted the protein expression of STAT-6 and GATA-3 considerably along with a modest reduction in p65-NF-κB phosphorylation. Furthermore, olaparib normalized the OVA-induced redox imbalance as reflected by data on reactive oxygen species, malondialdehyde, protein carbonyls, and reduced glutathione/oxidized glutathione ratio. Interestingly, the protection offered by olaparib was further linked with the altered level of NLRP3 inflammasome-mediated IL-1β release and consequent expression of its downstream targets matrix metalloproteinase-9 and transforming growth factor beta. Suppressed collagen deposition in the lungs correlates well with the reduced expression of vimentin upon olaparib treatment. Finally, olaparib restored the expression of histone deacetylase 2, a steroid-responsive element in asthma. Overall, results suggest that olaparib prevents OVA-induced airway inflammation as well as remodeling via modulating inflammasome signaling in mice. © 2019 IUBMB Life, 1-11, 2019.

Topics: Airway Remodeling; Animals; Apoptosis; Asthma; Cell Proliferation; Chronic Disease; Inflammasomes; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Phthalazines; Piperazines; Pneumonia; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Tumor Cells, Cultured

Synthetic therapeutic drugs for asthma, a chronic airway inflammation characterised by strong eosinophil, mast cell, and lymphocyte infiltration, mucus hyper-production, and airway hyper-responsiveness, exhibit numerous side effects. Alternatively, the high antioxidant potential of palm oil phytonutrients, including vitamin E (tocotrienol-rich fractions; TRF) and carotene, may be beneficial for alleviating asthma. Here, we determined the therapeutic efficacy of TRF, carotene, and dexamethasone in ovalbumin-challenged allergic asthma in Brown Norway rats. Asthmatic symptoms fully developed within 8 days after the second sensitization, and were preserved throughout the time course via intranasal ovalbumin re-challenge. Asthmatic rats were then orally administered 30 mg/kg body weight TRF or carotene. TRF-treated animals exhibited reduced inflammatory cells in bronchial alveolar lavage fluid. TRF- and carotene-treated rats exhibited notable white blood cell reduction comparable to that from dexamethasone. TRF- and carotene-treatment also downregulated pro-inflammatory markers (IL-β, IL-6, TNF-α), coincident with anti-inflammatory marker IL-4 and IL-13 upregulation. Treatment significantly reduced asthmatic rat plasma CRP and IgE, signifying improved systemic inflammation. Asthmatic lung histology displayed severe edema and inflammatory cell infiltration in the bronchial wall, whereas treated animals retained healthy, normal-appearing lungs. The phytonutrients tocotrienol and carotene thus exhibit potential benefits for consumption as nutritional adjuncts in asthmatic disease.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Carotenoids; Cytokines; Female; Male; Ovalbumin; Palm Oil; Rats; Tocotrienols

Several studies have reported that gut and lung microbiomes are involved in the process of asthma pathogenesis. However, it remains unclear how perinatal or early-life antibiotic intervention affect adult allergic airway inflammation. We assigned C57BL/6 mice randomly to four experimental groups: normal saline control (NS), ovalbumin (OVA), vancomycin pretreated NS (VAN-NS), and vancomycin pretreated OVA (VAN-OVA). The vancomycin groups were orally given the drug from gestational day 14 to 6 week. An OVA-induced asthma model was then established at 6 weeks of age, and airway inflammation was evaluated. In addition, total DNA was extracted from the feces and lung tissue and used for 16S rDNA gene sequencing, to detect the composition of the microbiome. In the VAN-OVA group, airway inflammation and Th2-related cytokines were found to be significantly increased versus the control groups. Gene sequencing showed that vancomycin treatment attenuated the richness and evenness, and altered the composition of the microbiome in the gut and lung.

Topics: Allergens; Animals; Animals, Newborn; Anti-Bacterial Agents; Asthma; Disease Models, Animal; Female; Homeostasis; Inflammation; Lung; Male; Mice; Mice, Inbred C57BL; Microbiota; Ovalbumin; Pregnancy; Prenatal Exposure Delayed Effects; RNA, Ribosomal, 16S; Vancomycin

Topics: Animals; Anti-Inflammatory Agents; Asthma; Dipsacaceae; Disease Models, Animal; Drugs, Chinese Herbal; Female; Immunoglobulin E; Interleukin-13; Interleukin-5; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Effect of alpha-linolenic acid (ALA) against ovalbumin (OVA)-induced inflammation, oxidant/antioxidant imbalance and pathological features was examined in rat.. Total and differential WBC count and oxidant/antioxidant levels in BALF (bronchoalveolar lavage fluid) as well as lung pathological features were investigated in five groups of rats including controls (group C), rats sensitized with OVA (group S) and S treated with either ALA (0.2 and 0.4 mg/ml) or dexamethasone.. As compared to group C, in OVA-sensitized rats, increases in WBC counts, levels of oxidant biomarkers and most pathological scores were observed while lymphocyte percentage and antioxidants levels decreased. Treatment with ALA (0.2 and 0.4 mg/ml) significantly reduced total WBC, NO. Alpha-linolenic acid suppressed inflammation and oxidative stress, making it a potential therapeutic candidate for treatment of airway inflammatory diseases such as bronchial asthma.

Topics: alpha-Linolenic Acid; Animals; Antioxidants; Asthma; Bronchoalveolar Lavage Fluid; Dexamethasone; Fibrosis; Inflammation; Leukocyte Count; Lung; Male; Ovalbumin; Rats; Rats, Wistar; Superoxide Dismutase

Ziziphora clinopodioides has been frequently used as an anti asthmatic plant in traditional medication. Recent work explores the anti-asthmatic activity of Z. clinopodioides in allergen-induced asthmatic mice. Intraperitoneal sensitization followed by intranasal challenge were given with ovalbumin (allergen) to develop allergic asthma. Investigational groups of animals were administered with drug methylprednisolone (MP) (15 mg/kg body weight), n-hexane fraction, ethylacetate fraction, and methanolic extract of Z. clinopodioides extract (500 mg/kg b.w.) for successive 07 days. Hematoxyline and eosin (H&E) and periodic acid-Schiff (PAS) stains were used to evaluate histopathological parameters on lung tissues. As an index of lungs tissues edema, wet/dry weight ratio of lungs was determined. Evaluation of expression levels of AQP1, AQP5, IL4, and IL5 was conducted by using RT-PCR. The data exhibited that both Z. clinopodioides and MP attenuated differential and total leukocyte counts in hematological examination i.e. in BALF and blood. Treatment with Z. clinopodioides also caused suppression of inflammatory cell infiltration and expression levels of IL4 and IL5, the later could have caused attenuation of pulmonary inflammation. The study also found decline in lung wet/dry ratio and goblet cellh hyperplasia in treated groups which indicates amelioration of lung edema. Treatment with Z. clinopodioides significantly increased the expression levels of aquaporin-1 and -5, which could have led to reduction in lung edema. The treatment with MP showed comparable results to Z. clinopodioides. Current investigation revealed that Z. clinopodioides possessed anti-asthmatic property which might be accredited to upregulagted AQP1 and AQP5 levels and downregulated IL4 and IL5 levels.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Aquaporins; Asthma; Cytokines; Disease Models, Animal; Down-Regulation; Female; Hypersensitivity; Inflammation; Mentha; Methylprednisolone; Mice; Ovalbumin; Plant Extracts; Pulmonary Edema; Up-Regulation

Glucagon has been shown to be beneficial as a treatment for bronchospasm in asthmatics. Here, we investigate if glucagon would prevent airway hyperreactivity (AHR), lung inflammation, and remodeling in a murine model of asthma. Glucagon (10 and 100 µg/Kg, i.n.) significantly prevented AHR and eosinophilia in BAL and peribronchiolar region induced by ovalbumin (OVA) challenge, while only the dose of 100 µg/Kg of glucagon inhibited subepithelial fibrosis and T lymphocytes accumulation in BAL and lung. The inhibitory action of glucagon occurred in parallel with reduction of OVA-induced generation of IL-4, IL-5, IL-13, TNF-α, eotaxin-1/CCL11, and eotaxin-2/CCL24 but not MDC/CCL22 and TARC/CCL17. The inhibitory effect of glucagon (100 µg/Kg, i.n.) on OVA-induced AHR and collagen deposition was reversed by pre-treatment with indomethacin (10 mg/Kg, i.p.). Glucagon increased intracellular cAMP levels and inhibits anti-CD3 plus anti-CD28-induced proliferation and production of IL-2, IL-4, IL-10, and TNF- α from TCD4

Topics: Airway Remodeling; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Cell Proliferation; Chemokine CCL24; Cytokines; Glucagon; Lung; Mice, Inbred Strains; Ovalbumin; Pneumonia; Receptors, Glucagon

Asthma is an allergic airway disease (AAD) characterized by eosinophilic inflammation, mucus hypersecretion, and airway hyper responsiveness, and it is caused by dysregulated immune responses. Conversely, regulatory T cells (Tregs) control aberrant immune responses and maintain homeostasis. Recent evidence suggests that Streptococcus pneumoniae, including its components as well as a live attenuated mutant, and pneumococcal infection induce Tregs and can thus potentially be harnessed therapeutically for asthma treatment. Previously, a pep27 deletion mutant (Δpep27) demonstrated a significantly attenuated virulence in a sepsis model, and Δpep27 immunization induced serotype-nonspecific protection against S. pneumoniae infection, as well as influenza virus, possibly via an immune tolerance mechanism. Here, the potential of Δpep27 immunization for asthma protection was studied. Mice were immunized intranasally with Δpep27 before or after ovalbumin sensitization and subsequent challenge. Δpep27 immunization suppressed hallmark features of AAD, including antigen-specific type 2 helper T cell cytokine and antibody responses, peripheral and pulmonary eosinophil accumulation, and goblet cell hyperplasia. Thus, a Δpep27 vaccine may be highly feasible as a preventive or therapeutic agent for asthma.

Topics: Administration, Intranasal; Animals; Asthma; Bacterial Proteins; Bronchoalveolar Lavage Fluid; Chronic Disease; Cytokines; Disease Models, Animal; Female; Mice, Inbred BALB C; Mutation; Ovalbumin; Pneumococcal Vaccines; Streptococcus pneumoniae; T-Lymphocytes, Regulatory; Th2 Cells

To investigate the effects of exposure to diethyl phthalate(DEHP) during pregnancy and lactation in respiratory allergy of offspring Wistar rats.. To establish the maternal DEHP exposure model: 36 healthy 2-month-old female Wistar rats were divided into three different dose groups. From GD0, each group of pregnant mice were given different concentrations of DEHP(0, 30, 300 mg/(kg·d)) until to the newborn weaning(PND21). After birth, one of offspring was selected from each cage in different dose groups. At PND21 and PND28 the offspring were sensitized by intraperitoneal injection(i. p. ) OVA and continuous nasal sensitization at PND32, PND33, PND34. Bronchoalveolar lavage fluid(BALF) and lung tissue were collected at PND35 with non-sensitized groups. ELISA detected the secretion of Th2 cytokine interleukin(IL)-4, IL-5 and IL-13 in BALF, and the pathological changes of allergic inflammation in lung tissues were observed by HE and PAS staining. Expression of epithelium-derived factor IL-33 detected by immunohistochemistry.. In BALF, compared with the control group, the total number of inflammatory cells and eosinophils in the DEHP0+OVA group increased to(131. 500±25. 548)×10~4, (32. 000±10. 079)×10~4(P<0. 05); The total number of inflammatory cells and eosinophils in the DEHP30+OVA group increased to(156. 167±17. 994)×10~4, (16. 331±6. 667)×10~4(P<0. 05); and the total number of inflammatory cells and eosinophils in the DEHP300+OVA group increased to(172. 167±19. 994)×10~4, (55. 000±17. 018)×10~4(P<0. 05). After adding different doses of DEHP and OVA, compared with the control group, The expression levels of IL-4, IL-5, IL-13 in DEHP0+OVA group increased to(38. 401±6. 594) pg/mL(P>0. 05), (30. 026±2. 756) pg/mL(P<0. 05), (13. 806±4. 355) pg/mL(P<0. 05); The expression levels of IL-4, IL-5 and IL-13 in DEHP30+OVA group increased to(57. 733±7. 293) pg/mL(P<0. 05) and(31. 544±1. 043) pg/mL(P>0. 05), (18. 068±1. 497) pg/mL(P<0. 05); The expression levels of IL-4, IL-5 and IL-13 in DEHP300+OVA group increased to(54. 943±6. 049) pg/mL(P>0. 05) and(32. 377±3. 739)pg/mL(P>0. 05), (20. 168±0. 939) pg/mL(P<0. 05), respectively. The tissue section of all the DEHP+saline groups could be observed that no obvious allergic inflammatory reaction, meanwhile all the DEHP+OVA groups had a relatively obvious allergic reaction and were severe with the increase of DEHP dose. Immunohistochemistry showed no significant increase in the expression of IL-33 in the DEHP+saline groups, while the expression of IL-33 in the DEHP+OVA groups increased with a certain dose response.. Exposure to DEHP during pregnancy and lactation will aggravate the sensitization reaction of offspring, the possible mechanism that DEHP increase the Th2 type of immune response may be the overexpression of IL-33 in the epithelial cells.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Humans; Hypersensitivity; Interleukin-33; Lactation; Mice; Mice, Inbred BALB C; Ovalbumin; Phthalic Acids; Pregnancy; Rats; Rats, Wistar

Siraitia grosvenorii fruits are used in traditional medicine to treat cough, sore throat, bronchitis, and asthma.. This study aimed to investigate the anti-inflammatory and anti-asthmatic effects of S. grosvenorii residual extract (SGRE) on ovalbumin (OVA)-induced asthma in mice.. Asthma was induced in BALB/c mice by systemic sensitization to OVA, followed by intratracheal, intraperitoneal, and aerosol allergen challenges. SGRE was orally administered for four weeks. We investigated the effects of SGRE on airway hyper-responsiveness, OVA-specific IgE production, histological analysis of lung and trachea, immune cell phenotyping, Th1/Th2 cytokine production in bronchoalveolar lavage fluid (BAL) fluid and splenocytes, and gene expression in the lung.. SGRE ameliorated OVA-driven airway hyper-responsiveness, serum IgE production, and histopathological changes in the lung and trachea. SGRE reduced the total number of cells in the lung and BAL, the total number of lymphocytes, neutrophils, monocytes, and eosinophils in the lung and BAL, the absolute number of CD4+/CD69+ T cells in the lung, and the absolute number of CD4+/CD8+ T cells and CD11b+/Gr-1+ granulocytes in the lung and BAL. SGRE also reduced Th2 cytokines (IL-4, IL-5, and IL-13) and increased the Th1 cytokine IFN-γ in the BAL fluid and supernatant of splenocyte cultures. SGRE decreased the OVA-induced increase of IL-13, TARC, MUC5AC, TNF-α, and IL-17 expression in the lung.. SGRE exerts anti-asthmatic effects via the inhibition of Th2 and Th17 cytokines and the increase of Th1 cytokines, suggesting that SGRE may be a potential therapeutic agent for allergic lung inflammation, such as asthma.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents, Non-Steroidal; Asthma; Bronchoalveolar Lavage Fluid; Cucurbitaceae; Disease Models, Animal; Down-Regulation; Eosinophils; Interleukins; Male; Mice, Inbred BALB C; Mucin 5AC; Ovalbumin; Plant Extracts; Pneumonia; Respiratory Hypersensitivity; Th17 Cells

Corticosteroids commonly prescribed in asthma show several side-effects. Relatively non-toxic andrographolide (AG) has an anti-asthmatic potential. But its poor bioavailability and short plasma half-life constrain its efficacy. To overcome them, we encapsulated AG in nanoparticle (AGNP) and evaluated AGNP for anti-asthmatic efficacy on murine asthma model by oral/pulmonary delivery. AGNP had 5.47% drug loading with a sustained drug release in vitro. Plasma and lung pharmacokinetic data showed predominantly improved AG-bioavailability upon AGNP administered orally/by pulmonary route. Cell numbers, IL-4, IL-5, and IL-13 levels in broncho-alveolar lavage fluid and serum IgE content were reduced significantly after administration of AGNP compared to free-AG treatment. AGNP-mediated suppression of NF-κβ was predominantly more compared to free-AG. Further, pulmonary route showed better therapeutic performance. In conclusion, AGNP effectively controlled mild and severe asthma and the pulmonary administration of AGNP was more efficacious than the oral route.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Diterpenes; Drug Liberation; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Male; Mice, Inbred C57BL; Nanoparticles; NF-kappa B; Ovalbumin; Particle Size; Rats, Sprague-Dawley; Signal Transduction; Spectroscopy, Fourier Transform Infrared; Tissue Distribution

Asthma is a chronic inflammatory disease of the airways of the lungs. Pomalidomide (POM) a therapy for multiple myeloma has been stated to have an anti-inflammatory effect. The main goal of the present study was to assess its possible effect on airway contraction and inflammation in a rat model of ovalbumin-induced asthma. Different groups of rats received saline or pomalidomide (0.4, 0.8 mg/kg) or dexamethasone (0.6 mg/kg). The asthma was induced by ovalbumin (OVA). Trachea contraction was assayed by organ bath system. Airway histology was assessed using hematoxylin and eosin method. Serum Tumor necrosis factor alpha (TNF-α) level was analyzed by Enzyme-Linked Immunosorbent Assay and Platelet-derived growth factor (PDGFα) Gene expressions were evaluated by Real-time PCR. Pomalidomide prevented ovalbumin-induced airway contraction and histopathological damage. In addition serum, TNF-α level was significantly (p<0.05) decreased in POM treated animals compared to control (asthmatic animals that received POM vehicle). Results indicate that POM prevented the PDGF expression induced by ovalbumin. In conclusion, we found that pomalidomide ameliorated the symptoms, histopathological changes and inflammatory markers induced by ovalbumin in asthmatic rats and these effects might be related to its anti-inflammatory properties.

Topics: Airway Obstruction; Allergens; Animals; Anti-Inflammatory Agents; Asthma; Disease Models, Animal; Humans; Lung; Male; Ovalbumin; Platelet-Derived Growth Factor; Rats; Rats, Wistar; Thalidomide; Tumor Necrosis Factor-alpha

Asthma is characterized by airway inflammation and mucus hypersecretion. Curcumin possessed a potent anti-inflammatory property involved in the PPAR

Topics: Animals; Asthma; Blotting, Western; Cell Line; Curcumin; Humans; Interleukin-13; Interleukin-4; Interleukin-5; Male; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; PPAR gamma; RNA, Small Interfering; Signal Transduction; Tumor Necrosis Factor-alpha

BACKGROUND The purpose of the present study was to investigate the function and mechanism of dihydroartemisinin (DHA) in treating ovalbumin-induced asthma in BALB/c mice. MATERIAL AND METHODS Thirty female BALB/c mice were randomly separated into 3 groups: the control group, the asthma model group stimulated by ovalbumin (OVA group), and the DHA treatment group (DHA group). The therapeutic effects and potential pharmacological mechanisms of DHA were specifically clarified by examining its effects on asthma-related phenomena, such as body weight, lung function, cell counts in bronchoalveolar lavage fluid (BALF), and hemotoxin and eosin staining. In addition, the expression of inflammatory factors was checked by enzyme-linked immunosorbent assay kits, and fractions of Th17 cells were detected by FACS analysis. Moreover, the downstream molecular pathway of IL-6/Stat3 (interleukin-6/signal transducer and activator of transcription 3) and expression of miR-183C was investigated by western blot and/or quantitative real-time polymerase chain reaction. Luciferase assay was used to reveal the function of miR-183C on the transcriptional regulation of Foxo1 (forkhead box O). RESULTS DHA administration significantly relieved the severity of the asthma through its effect on body weight, survival rate, and airway pressure. DHA was able to ameliorate lung damage in terms of pathological morphology and it reduced the percentage of helper T 17 (Th17) cells and the secretion of cytokines. As a result, the activity of the IL-6/Stat3 pathway was inhibited by DHA. In addition, the adoption of DHA decreased the expression of miR-183C but increased the expression of the transcription factor Foxo1. CONCLUSIONS Our results suggest that the therapeutic effects of DHA on asthma are partially realized via the regulation of miR-183C and IL-6/Stat3 pathway.

Topics: Allergens; Animals; Artemisinins; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Forkhead Transcription Factors; Lung; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; STAT3 Transcription Factor; T-Lymphocytes, Regulatory; Th17 Cells

Asthma is a chronic inflammatory disease that involves a variety of cytokines and cells. Interleukin-16 (IL-16) is highly expressed during allergic airway inflammation and is involved in its development. However, its specific mechanism of action remains unclear. In the present study, we used an animal model of ovalbumin (OVA)-induced allergic asthma with mice harboring an IL-16 gene deletion to investigate the role of this cytokine in asthma, in addition to its underlying mechanism. Increased IL-16 expression was observed during OVA-induced asthma in C57BL/6J mice. However, when OVA was used to induce asthma in IL-16

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Disease Models, Animal; Female; Immunoglobulin E; Interleukin-16; Lung; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Signal Transduction; Spleen; Th17 Cells; Th2 Cells

The present study aims to investigate the effects of toll-like receptor 4 (TLR4) antagonist in an ovalbumin (OVA)-induced mouse model of combined allergic rhinitis and asthma syndrome (CARAS). An OVA-induced mouse model of CARAS was established and TLR4 antagonist, TAK-242, was administrated intranasally or intraperitoneally. The number of sneezing and nasal rubbing was counted. The frequency of different cell types in the bronchoalveolar lavage fluid (BALF) and nasal lavage fluid (NLF) was analyzed using flow cytometry. Expressions of protein in nasal mucosa and lungs were determined using western blotting. Levels of interleukin (IL)-4, IL-5, and IL-13 were determined using Enzyme-linked Immunosorbent Assay (ELISA). Histological scores were applied for the assessment of lung injury. Treatment of TAK-242 downregulated CCL2 expression and reduced monocyte infiltration in nasal mucosa and lung tissues. Additionally, treatment of TAK-242 ameliorated upper airway symptoms including the sneezing and nasal rubbing by the regulation of cytokines including IL-4, IL-5, and IL-13. Furthermore, treatment of TAK-242 ameliorated lower airway symptoms including decreasing the frequency of CD45

Topics: Animals; Anti-Inflammatory Agents; Asthma; Cytokines; Disease Models, Animal; Female; Lung; Mice, Inbred BALB C; Monocytes; Nasal Mucosa; Ovalbumin; Rhinitis, Allergic; Sulfonamides; Syndrome; Toll-Like Receptor 4

Asthma is a chronic inflammatory lung disease of the airway; the incidence and prevalence of asthma remain high worldwide. Astragaloside IV (AS-IV) is the main active constituent of Astragalus membranaceus. Accumulating evidence suggests that AS-IV possesses anti-inflammatory and anti-asthmatic ability, but the potential molecular mechanism is required to further clarify. In this study, the anti-asthmatic effects of AS-IV on mice with ovalbumin (OVA)-induced allergic inflammation were analysed. We analysed airway hyperresponsiveness (AHR), numbers of inflammatory cells, inflammation situation in lung tissue and cytokines level in bronchoalveolar lavage fluid (BALF) between OVA-induced mice with and without AS-IV treatment. Moreover, we explored the possible signalling pathway behind the anti-asthmatic effects. Our results revealed that AS-IV treatment ameliorates airway inflammation and AHR in an OVA-induced asthma model. Besides, AS-IV treatment inhibits the interleukin (IL)-4, -5 and -13 production, and further study indicated that AS-IV treatment downregulates the expression level of p-JAK2/p-STAT6 proteins. Taken together, the present study suggested that the inhibitory effects of AS-IV on asthma therapy are at least partially involved in inhibiting the JAK2/STAT6 signalling pathway.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Female; Gene Expression Regulation; Janus Kinase 2; Leukocytes; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Parasympathomimetics; Saponins; Signal Transduction; STAT6 Transcription Factor; Triterpenes

Asthma is a chronic disease with impacts on public health. It affects the airways causing pulmonary inflammation mediated by CD4 T cells type Th2, eosinophilia, mucus hypersecretion, and elevated IgE. The unbalance between cytokines and transcription factors is an important feature in asthma. Probiotics has gaining highlight as a therapy for chronic diseases. Thus, we investigate the Lactobacillus bulgaricus (Lb) effect in murine allergic asthma. BALB/c-mice were sensitized to ovalbumin (OA) on days 0 and 7 and were challenged from day 14-28 with OA. Mice received Lb seven days prior to sensitization and it was kept until day 28. The Lb attenuated the eosinophils infiltration, mucus and collagen secretion, IgE production, pro-inflammatory cytokines, TLR4 expression, GATA3, STAT6 and RORγt in lung. Otherwise, Lb increased the anti-inflammatory cytokines, the T-bet and foxp3. Finally, Lb attenuated the allergic asthma-induced inflammation and airway remodeling by interfering on Th1/Th2 cytokines and STAT6/T-bet transcription factors.

Topics: Airway Remodeling; Animals; Asthma; Cytokines; Disease Models, Animal; Eosinophils; GATA3 Transcription Factor; Gene Expression Regulation; Immunoglobulin E; Lactobacillus delbrueckii; Male; Mice; Mice, Inbred BALB C; Nuclear Receptor Subfamily 1, Group F, Member 3; Ovalbumin; Pneumonia; Probiotics; Signal Transduction; STAT6 Transcription Factor; T-Box Domain Proteins; Th1 Cells; Th1-Th2 Balance; Th2 Cells; Toll-Like Receptor 4

In asthma, there are high levels of inflammatory mediators, reactive oxygen species (ROS), and eosinophil extracellular traps (EETs) formation in airway. Here, we attempted to investigate the ROS involvement in EETs release and airway inflammation in OVA-challenged mice. Before the intranasal challenge with ovalbumin (OVA), animals were treated with two ROS inhibitors, N-acetylcysteine (NAC) or diphenyleneiodonium (DPI). We showed that NAC treatment reduced inflammatory cells in lung. DPI and NAC treatments reduced eosinophil peroxidase (EPO), goblet cells hyperplasia, proinflammatory cytokines, NFκB p65 immunocontent, and oxidative stress in lung. However, only the NAC treatment improved mitochondrial energy metabolism. Moreover, the treatments with DPI and NAC reduced EETs release in airway. This is the first study to show that ROS are needed for EETs formation in asthma. Based on our results, NAC and DPI treatments can be an interesting alternative for reducing airway inflammation, mitochondrial damage, and EETs release in asthma.

Topics: Acetylcysteine; Animals; Asthma; Cytokines; Energy Metabolism; Eosinophil Peroxidase; Eosinophils; Extracellular Traps; Female; Goblet Cells; Lung; Mice; Mice, Inbred BALB C; Mitochondria; Onium Compounds; Ovalbumin; Oxidative Stress; Reactive Oxygen Species; Transcription Factor RelA

Maintaining an airway clear of bacteria, foreign particles and apoptotic cells by alveolar macrophages is very essential for lung homeostasis. In asthma, the phagocytic capacity of alveolar macrophages is significantly reduced, which is thought to be associated with increased oxidative stress. Hydrogen (H. Female C57BL/6 mice were intraperitoneally sensitized with OVA before they were subject to airway challenge with aerosolized OVA. Hydrogen gas was delivered to the mice through inhalation twice a day (2 h once) for 7 consecutive days. Phagocytic function of alveolar macrophages isolated from bronchoalveolar lavage fluid was assessed by fluorescence-labeled Escherichia coli as well as flow cytometry.. Alveolar macrophages isolated from OVA-induced asthmatic mice showed decreased phagocytic capacity to Escherichia coli when compared with those of control mice. Defective phagocytosis in asthmatic mice was reversed by hydrogen gas inhalation. Hydrogen gas inhalation significantly alleviated OVA-induced airway hyperresponsiveness, inflammation and goblet cell hyperplasia, diminished T. Our findings demonstrated that hydrogen gas inhalation enhanced alveolar macrophage phagocytosis in OVA-induced asthmatic mice, which may be associated with the antioxidant effects of hydrogen gas and the activation of the Nrf2 pathway.

Topics: Administration, Inhalation; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Escherichia coli; Female; Heme Oxygenase-1; Hydrogen; Lung; Macrophages, Alveolar; Membrane Proteins; Mice, Inbred C57BL; NF-E2-Related Factor 2; NF-kappa B; Ovalbumin; Phagocytosis

Asthma is the most common chronic airway inflammatory disease. Sirtuin 1 (SIRT1) exerts a crucial effect on regulating chronic inflammatory responses. Therefore, this study aims to explore the effect of SIRT1 on the pathogenesis of asthma.. Serum level of SIRT1 in asthma patients and healthy controls was detected by Western blot. Correlation between SIRT1 level and pulmonary function in asthma patients was analyzed. Subsequently, asthma model in mouse was established. Primary airway epithelial cells were extracted from asthma mice and control mice to detect SIRT1 level. Furthermore, relative levels of Akt and interleukin 6 (IL-6) were detected in 16HBE cells. Regulatory effects of Akt on SIRT1 in 16HBE cells were determined as well.. SIRT1 was highly expressed in serum of asthma patients, which was negatively correlated with FEV1/FVC (r=-0.27, **p<0.01). Both mRNA and protein levels of SIRT1 were downregulated in primary airway epithelial cells extracted from asthma mice compared with those from controls. SIRT1 knockdown in 16HBE cells upregulated IL-6 expression, which was reversed by Akt inhibitors.. SIRT1 regulates IL-6 level via Akt pathway, thereafter affecting pulmonary function in asthma patients.

Topics: Animals; Asthma; Cell Line; Disease Models, Animal; Epithelial Cells; Female; Gene Knockdown Techniques; Humans; Interleukin-6; Leukocyte Count; Male; Mice; Neutrophils; Ovalbumin; Primary Cell Culture; Proto-Oncogene Proteins c-akt; Respiratory Function Tests; Severity of Illness Index; Signal Transduction; Sirtuin 1; Up-Regulation

Particulate matter (PM) exposure increases the risk of asthma. However, the effect of PM

Topics: Allergens; Animals; Antioxidants; Asthma; Gas Chromatography-Mass Spectrometry; Liver; Male; Metabolic Networks and Pathways; Mice; Ovalbumin; Particulate Matter

Topics: Animals; Antibody Specificity; Asthma; Bronchoalveolar Lavage Fluid; Cell Adhesion; Chalcones; Chemokines; Collagen; Cyclooxygenase 2; Disease Models, Animal; DNA Damage; Eosinophils; Female; Glutathione; Goblet Cells; Humans; Hyperplasia; Inflammation Mediators; Lung; Malondialdehyde; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Protective Agents; Reactive Oxygen Species; Respiratory Hypersensitivity; THP-1 Cells

We previously reported that the nonsteroidal compound CpdX, which was initially characterized 20 y ago as a possible gestagen and, shortly afterward, as a possible drug for treatments of inflammatory diseases, selectively triggers the NFκB/AP1-mediated tethered indirect transrepression function of the glucocorticoid receptor (GR), and could therefore be a selective glucocorticoid receptor agonistic modulator (SEGRAM). We now demonstrate that, upon administration to the mouse, CpdX and one of its deuterated derivatives, CpdX-D3, repress as efficiently as a synthetic glucocorticoid (e.g., Dexamethasone) an induced skin atopic dermatitis, an induced psoriasis-like inflammation, a house dust mite (HDM)-induced asthma-like allergic lung inflammation, a collagen-induced arthritis, an induced ulcerative colitis, and an ovalbumin-induced allergic conjunctivitis. Interestingly, in the cases of an HDM-induced asthma-like allergic lung inflammation and of a collagen-induced arthritis, the CpdX antiinflammatory activity was selectively exerted by one of the two CpdX enantiomers, namely, CpdX(eA) or CpdX-D3(eA).

Topics: Animals; Anti-Inflammatory Agents; Arthritis, Experimental; Asthma; Conjunctivitis, Allergic; Dermatitis, Atopic; Dexamethasone; Disease Models, Animal; Glucocorticoids; Humans; Inflammation; Mice; NF-kappa B; Ovalbumin; Progestins; Receptors, Glucocorticoid; Skin; Transcriptional Activation

Topics: Administration, Inhalation; Animals; Asthma; Bronchoalveolar Lavage Fluid; Bronchoconstriction; Bronchodilator Agents; Cytokines; Disease Models, Animal; Eosinophils; Immunoglobulin E; Lung; Male; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Stereoisomerism; Terbutaline

Recently, we demonstrated that Rac1 upregulation is involved in augmented bronchial smooth muscle (BSM) contractions of antigen-challenged mice. However, change in G protein-coupled receptor (GPCR)-induced Rac1 activation remains unknown in BSMs of repeatedly antigen-challenged (Chal.) mice. We here examined carbachol (CCh)-induced Rac1 activation in BSMs of Chal. mice. Gene expression levels of both Rac1 and Rac-guanine nucleotide exchange factors (GEFs), such as Tiam1 and Trio, were increased in BSMs of Chal. mice. Furthermore, CCh-induced Rac1 activation was inhibited by pretreatment with Rac1-GEF inhibitor NSC23766 and Rac1 inhibitor EHT1864 in BSMs of sensitized-control (S.C.) and Chal. mice. Compared with S.C. mice, CCh-induced Rac1 activation was increased in BSMs of Chal. mice. In conclusion, we reported that increased CCh-induced Rac1 activation via Tiam1 and Trio upregulation, in addition to upregulate Rac1, may be involved in increased CCh-induced BSM contractions in Chal. mice.

Topics: Aminoquinolines; Animals; Antigens; Asthma; Bronchi; Carbachol; Guanine Nucleotide Exchange Factors; Male; Mice, Inbred BALB C; Muscarinic Agonists; Muscle Contraction; Muscle, Smooth; Neuropeptides; Ovalbumin; Phosphoproteins; Protein Serine-Threonine Kinases; Pyrimidines; Pyrones; Quinolines; rac1 GTP-Binding Protein; T-Lymphoma Invasion and Metastasis-inducing Protein 1; Up-Regulation

Helminths immunomodulate the host immune system by secreting proteins to create an inhibitory environment as a strategy for survival in the host. As a bystander effect, this balances the host immune system to reduce hypersensitivity to allergens or autoantigens. Based on this, helminth therapy has been used to treat some allergic or autoimmune diseases. As a tissue-dwelling helminth, Trichinella spiralis infection has been identified to have strong immunomodulatory effects; the effective components in the worm have not yet been identified.. The soluble extracts of T. spiralis adult worms and muscle larvae were used to treat airway inflammation before and after an ovalbumin (OVA)-sensitization/challenge in an OVA-induced asthma mouse model. The therapeutic effects were observed by measuring the level of inflammation in the lungs.. The soluble products derived from T. spiralis parasites, especially from adult worms, were able to ameliorate OVA-induced airway inflammatory responses which were associated with reduced eosinophil infiltration, OVA-specific IgE, Th2 cytokine IL-4, and increased IL-10 and TGF-β. The stimulation of the Treg response may contribute to the alleviated allergic inflammation.. Trichinella spiralis worm extracts stimulate regulatory cytokines that are associated with reduced allergic airway inflammation. The identification of effective components in the adult worm extracts will be a crucial approach for developing a novel therapeutic for allergic and autoimmune diseases.

Topics: Animals; Anti-Allergic Agents; Asthma; Cytokines; Disease Models, Animal; Eosinophils; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells; Tissue Extracts; Trichinella spiralis

Linalool is a natural product present in fruits and aromatic plants with biological activities. Researchers have reported that the inhalation of linalool exerts anti-inflammatory activities. In this study, we examined the therapeutic effects of linalool on airway inflammation and mucus overproduction in mice with allergic asthma. Oral administration of linalool significantly inhibited the levels of eosinophil numbers, Th2 cytokines and immunoglobulin E (IgE) caused by ovalbumin (OVA) exposure. Linalool exerted preventive effects against the influx of inflammatory cells and mucus hypersecretion in the lung tissues. Linalool also dose-dependently decreased the levels of inducible nitric oxide synthase (iNOS) expression and protein kinase B (AKT) activation in the lung tissues. Linalool effectively downregulated the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor-κB (NF-κB) caused by OVA exposure. Furthermore, linalool exerted inhibitory effect on OVA-induced airway hyperresponsiveness (AHR). In the in vitro study, the increased secretion of MCP-1 was attenuated with linalool treatment in lipopolysaccharide (LPS)-stimulated H292 airway epithelial cells. In conclusion, linalool effectively exerts a protective role in OVA-induced airway inflammation and mucus hypersecretion, and its protective effects are closely related to the downregulation of inflammatory mediators and MAPKs/NF-κB signaling.

Topics: Acyclic Monoterpenes; Administration, Oral; Allergens; Animals; Anti-Inflammatory Agents; Asthma; Cells, Cultured; Cytokines; Disease Models, Animal; Female; Humans; Hypersensitivity; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; NF-kappa B; Nitric Oxide Synthase Type II; Ovalbumin; Respiratory Hypersensitivity; Respiratory Mucosa; Th2 Cells

Orosomucoid-like protein 3 (ORMDL3) is a common mutation in many asthma patients and its effects on the specific pathogenesis of asthma are still unclear. Therefore, in this study, we used a mouse that specifically knockout the mouse ORDML3 gene to further study the mechanism. We used ovalbumin (OVA) to induce asthma in wild-type mice and ORMDL3 knockout mice. Lung ventilation resistance, airway inflammation, mucus hypersecretion, collagen deposition, the levels of inflammatory factors and the expression of ORDML3 and JNK1/2-MMP-9 pathway were detected. The results showed that ORMDL3 gene was highly expressed in clinical asthmatic children and mouse asthma model. Knocking down the ORMDL3 gene in the lung tissue of asthmatic mice can reduce airway hyperresponsiveness, airway inflammation, mucus secretion, and collagen deposition around the airway. After knocking down the lung tissue of mice, the IL-4, IL-5 and IL-13 concentrations in broncho alveolar lavage fluid of asthmatic mice were significantly decreased, and the activation of JNK1/2-MMP-9 pathway was inhibited in mouse lung tissue. Collectively, our results demonstrate that the ORMDL3 gene may aggravate asthma symptoms by activating the JNK1/2-MMP-9 pathway, which indicates that the ORMDL3 gene may be the key molecule for the next step of asthma targeted therapy.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Child; Disease Models, Animal; Female; Gene Expression Regulation; Humans; Interleukin-13; Interleukin-4; Interleukin-5; Lung; Matrix Metalloproteinase 9; Membrane Proteins; Mice; Mice, Inbred C57BL; Mice, Knockout; Mitogen-Activated Protein Kinase 8; Mitogen-Activated Protein Kinase 9; Mucus; Ovalbumin; Respiratory Function Tests; Respiratory Hypersensitivity; Signal Transduction

This study detected the expressions of microRNA-26a (miR-26a), miR-146a and miR-31 in lung tissues and BALF (bronchoalveolar lavage fluid) of asthma mice and children. Besides, cytokine levels of interleukin-5 (IL-5), IL-8, IL-12 and tumor necrosis factor-α (TNF-α) were detected as well. We aim to provide an experimental basis for clinical treatment of asthma.. Forty female BALB/c mice were randomly assigned into control group and asthma group, respectively. Mice in asthma group (n=20) were immunized by intraperitoneal injection of OVA (ovalbumin) and provoked by atomization inhalation of OVA from the 15th day for 10 days. Mice in control group (n=20) were immunized and provoked with isodose saline during the same period. At the 26th day, mice were sacrificed for collecting lung tissues and BALF. Besides, we enrolled 17 cases of asthma children and 13 cases of children with airway foreign body as controls. BALF of each subject was collected. Total cellular score and differential counting in BALF were recorded. Expression levels of miR-26a, miR-146a, and miR-31 were detected by reverse transcription-polymerase chain reaction (RT-PCR). Levels of IL-5, IL-8, IL-12, and TNF-α were detected by enzyme-linked immunosorbent assay (ELISA).. The total cellular score in BALF of asthma mice and asthma children was higher than that of controls (p<0.05). Percentages of eosinophils, neutrophils, and lymphocytes in BALF of asthma mice and asthma children were higher than those of controls, whereas the percentage of macrophages was lower (p<0.05). Levels of IL-5, IL-8, IL-12, and TNF-α in lung tissues of asthma mice were markedly elevated compared with those of controls (p<0.05). Similarly, levels of IL-5, IL-8, IL-12, and TNF-α were higher in BALF of asthma children than controls (p<0.05). RT-PCR data showed higher mRNA levels of miR-26a, miR-146a, and miR-31 in lung tissues of asthma mice than controls (p<0.05). The mRNA levels of miR-26a, miR-146a, and miR-31 in BALF of asthma children were highly expressed compared with those of controls as well (p<0.05).. MiR-26a, miR-146a, and miR-31 are involved in asthma progression mainly through regulating inflammatory factors and cells.

Topics: Adolescent; Animals; Asthma; Bronchoalveolar Lavage Fluid; Case-Control Studies; Disease Models, Animal; Disease Progression; Eosinophils; Female; Foreign Bodies; Humans; Inflammation Mediators; Lung; Lymphocytes; Male; Mice; MicroRNAs; Neutrophils; Ovalbumin; Up-Regulation; Young Adult

Asthma is a chronic, complex and heterogeneous inflammatory illness, characterized by obstruction of the lower airways. About 334 million people worldwide suffer from asthma, and these estimates, as well as the severity of the disease, have increased in the last decades. Glucocorticoids are currently the most widely used drugs in the treatment and control of asthma symptoms, but their prolonged use can cause serious adverse effects. N-acylhydrazone derivatives have been tested in pre-clinical studies in models of inflammatory diseases. Here we tested SintMed65 (N'-[(1E)-3-(4-nitrophenylhydrazono)]-(2E)-propan-2-ylidene-3,5-dinitrobenzohydrazide), a compound belonging to a novel class of immunosuppressive drugs, in a mouse model of allergic airway inflammation. BALB/c mice were sensitized previously and challenged with ovalbumin for five consecutive days and SintMed65 treatment was performed orally 1 h prior to challenge with ovalbumin. Administration of SintMed65, as well as the reference drug dexamethasone, reduced cellularity and the number of eosinophils in the bronchoalveolar fluid (BALF). SintMed65 also reduced the production of Th2 cytokines IL-4, IL-5 and IL-13 in the BALF, and IL-4, IL-10 and CCL8 gene expression in lung, compared to vehicle-treated mice. Importantly, a reduction in the number of leukocytes and in the mucus production in lungs of SintMed65-treated mice was found, compared to the vehicle-treated group. In contrast, IgE production was not significantly altered after treatment with SintMed65. Our results demonstrate that compound SintMed65 possesses anti-inflammatory characteristics, suggesting its therapeutic potential for the treatment of allergic diseases.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Hydrazones; Leukocyte Count; Lung; Male; Mice, Inbred BALB C; Mucus; Ovalbumin

Topics: Animals; Antitussive Agents; Asthma; Capsules; Cough; Disease Models, Animal; Drugs, Chinese Herbal; Endoplasmic Reticulum Stress; Inflammasomes; Male; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Rats, Wistar; Respiratory Function Tests

Lepidii seu Descurainiae Semen (LDS) is used as a traditional herbal medicine in northeast Asia, mainly in Korea, Japan, and China to treat lung disorders including coughs and phlegm caused by acute and chronic airway inflammation.. Recently, interest regarding health problems incurred by air pollution has rapidly grown. Herbal medicines are being considered as alternative agents to treat various diseases. In the present study, we evaluated and compared the anti-inflammatory effects of LDS, which is derived from Lepidium apetalum Willd. extracts (LAE) and Descurainia sophia (L.) Webb ex Prantl extracts (DSE), on allergic airway inflammation.. We established an ovalbumin-induced asthmatic mouse model to evaluate the efficacy of LDS extracts. We performed histological examination and measured relevant inflammatory mediators and cells in bronchoalveolar lavage fluid and lung. Furthermore, we conducted an in vitro T helper 2 (Th2) polarization assay, flow cytometry, and western blot analysis.. Asthmatic phenotypes were attenuated by LDS extract treatments. LDS extract administration significantly reduced mucus production, inflammatory cell infiltration into airways, and eosinophil activation. Furthermore, LDS extracts reduced the expression of type 2 cytokines and inhibited differentiation and activation of Th2 cells.. LDS alleviated eosinophilic inflammation by inhibiting Th2 cell differentiation, and DSE was more effective in attenuating allergic lung inflammation than LAE.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Brassicaceae; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Cytokines; Eosinophils; Female; Lung; Mice, Inbred BALB C; Ovalbumin; Plant Extracts

BACKGROUND This study investigated the effects and underlying mechanisms of emodin on cough variant asthma (CVA) in mice. MATERIAL AND METHODS The bronchial asthma mouse model was successfully established by use of ovalbumin (OVA) sensitization and challenge. The BALB/c mice were divided into 6 groups: a control group, an OVA model without or with emodin (15, 30, 60 mg/kg) group, and a dexamethasone (0.5 mg/g) group. The effect of the treatment was determined by measuring airway responsiveness. The levels of immunoglobulin molecules, as well as inflammatory cytokines in bronchoalveolar lavage fluid (BALF) and serum, were determined by ELISA. The lung tissues were stained by hematoxylin-eosin (HE). The expressions of Notch receptors (Notch 1-3) and Delta-like (DLL) 4 in the lung tissues were detected by RT-PCR and Western blot analysis. RESULTS Compared with the model group, emodin treatment significantly increased the levels of immunoglobulin E (IgE) and IgG1/IgG2a in BALF and serum (p<0.05). HE results indicated that emodin inhibited the infiltration of inflammatory cells and that emodin reduced the levels of inflammatory cytokines, interleukin (IL)-5, IL-17, and interferon (IFN)-γ in BALF and serum (p<0.05). Furthermore, the expressions of Notch 1, 2, 3, and DLL4 in lung tissue were inhibited by emodin treatment. CONCLUSIONS The results demonstrated that emodin alleviated inflammation in CVA mice, which might be associated with suppression of the Notch pathway. Emodin might be a promising therapeutic agent for allergic asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cough; Cytokines; Disease Models, Animal; Emodin; Eosinophils; Immunoglobulin E; Immunoglobulin G; Inflammation; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Notch

Paeonia lactiflora Pall. (peony) is a medicinal plant used in the Xiaoqinglong decoction, a commonly prescribed traditional Chinese medicine for asthma. The main active ingredients of peony roots-described as the total glucosides of peony (TGP)-have anti-inflammatory, immunomodulatory, and protective effects on endothelial cells, and they are known to improve rheumatoid arthritis. This study explored the underlying mechanism of TGP activity in the treatment of allergic asthma.. Allergic asthma was induced in BALB/c mice by administering injections of ovalbumin (OVA) mixed with aluminum hydroxide gel and inhaling nebulized OVA. The OVA-sensitized mice were treated with TGP by oral gavage, and the potentially anti-asthmatic treatment effect was studied by testing airway hyperresponsiveness, classifying and counting of leukocytes, performing cytokine assays, and analyzing the lung histopathology. The β-hexosaminidase activity was assayed as a biomarker to evaluate the effect of TGP on mast cell degranulation. The mechanism of TGP was explored by monitoring the Ca. In mice with OVA-induced allergic asthma, TGP reduced airway hyperresponsiveness and improved lung tissue pathology, which included a decrease in inflammatory cell infiltration and collagen deposition. TGP also significantly lowered BALF leukocyte, eosinophil, and neutrophil counts, along with chemokines and cytokines, such as eotaxin, TNF-α, IL-4, and MIP-1α, in serum and lungs of OVA-challenged mice. These effects were further confirmed with the decrease of β-hexosaminidase release and the inhibition of Ca. Our findings suggest that TGP improved OVA-induced allergic asthma in mice mainly by suppressing Ca

Topics: Animals; Anti-Asthmatic Agents; Asthma; beta-N-Acetylhexosaminidases; Bronchoalveolar Lavage Fluid; Calcium; Cell Degranulation; Cell Line, Tumor; Cytokines; Glucosides; Leukocyte Count; Male; Mast Cells; Mice, Inbred BALB C; Ovalbumin; Paeonia; Rats

Mangiferin is the major bioactive ingredient in the leaves of Mangifera indica L., Aqueous extract of such leaves have been traditionally used as an indigenous remedy for respiratory diseases including cough and asthma in Traditional Chinese Medicine. Mangiferin was shown to exert its anti-asthmatic effect by modulating Th1/Th2 cytokines imbalance via STAT6 signaling pathway. However, compelling evidence indicated that subtypes of T helpers and regulatory T cells other than Th1/Th2 were also involved in the pathogenesis of asthma. In current study, we investigated the effects of mangiferin on the differentiation and function of Th9, Th17 and Treg cells in a chicken egg ovalbumin (OVA)-induced asthmatic mouse model. Mangiferin significantly attenuated the symptoms of asthma attacks, reduced the total number of leukocytes, EOS and goblet cells infiltration in lung. Simultaneously, treatment with mangiferin remarkably decreased the proportion of Th9 and Th17 cells; reduced the levels of IL-9, IL-17A; inhibited the expression of PU.1 and RORγt in lung. However, the proportion of Treg cells, the expression of IL-10, TGF-β1 and Foxp3 were increased by mangiferin. Our data suggest that mangiferin exerted anti-asthmatic effect through decreasing Th9 and Th17 responses and increasing Treg response in OVA-induced asthmatic mouse model.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Egg Hypersensitivity; Female; Lung; Mangifera; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Signal Transduction; T-Lymphocytes, Regulatory; Th17 Cells; Th2 Cells; Xanthones

In allergic asthma, regulatory T cell (Treg) number and function are decreased. Antigen-primed CD8. Female C57BL/6 mice were used to develop the model of allergic asthma. Experimental mice were immunized with ovalbumin (OVA) by intra-peritoneal (i.p) injection and then challenged with OVA by intra-tracheal administration. Control mice were immunized with vehicle by i.p injection and challenged with OVA. Airway inflammation was determined by histology and AHR was measured by an invasive method. Levels of interferon (IFN)-γ, IL-4, and IL-17 in bronchoalveolar lavage fluid (BALF) were determined by enzyme-linked immunosorbent assay. The frequencies of CD8. Experimental mice displayed phenotypes of allergic asthma, including inflammatory cell infiltration into the lungs, goblet cell hyperplasia, increased airway resistance, and increased IL-4 and IL-17 in BALF. Compared to control mice, experimental mice displayed lower CD39. In allergic asthma, increased Tc2 and Tc17 are possibly related to insufficient CD39

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; CD8-Positive T-Lymphocytes; Disease Models, Animal; Female; Forkhead Transcription Factors; Humans; Interleukin-17; Interleukin-4; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; T-Lymphocytes, Regulatory

The present investigation aimed to evaluate the activity of the essential oil of Mentha arvensis L. on exogenously induced bronchoconstriction in experimental animals. The anti-asthmatic effect of M. arvensis essential oil (MAEO) was studied using histamine aerosol-induced bronchoconstriction in guinea pigs and ovalbumin (OVA) sensitised albino mice. Treatment with M. arvensis oil significantly (p < 0.001) increased the time of preconvulsive dyspnoea in histamine-induced guinea pigs. Oral treatment of MAEO significantly (p < 0.001) decreased absolute eosinophil count, serum level of IgE and the number of eosinophils, neutrophils in BALF. Histopathological examination of lungs showed that essential oil rescinded bronchial asthma. The present investigation provides evidence that MAEO relaxes bronchial smooth muscles and suppressed immunological response to OVA.

Topics: Animals; Asthma; Bronchoconstriction; Disease Models, Animal; Dyspnea; Eosinophils; Female; Guinea Pigs; Histamine; Immunoglobulin E; Lung; Mentha; Mice; Neutrophils; Oils, Volatile; Ovalbumin

This study aims to explore the influences of Paraoxonase-1 (PON1) involved in airway inflammation and remodeling in asthma. Mice were divided into control, asthma, asthma + PON1 and asthma + NC groups, and asthma models were established via aerosol inhalation of ovalbumin (OVA). HE, Masson, and PAS stains were used to observe airway inflammation and remodeling, Giemsa staining to assess inflammatory cells in bronchoalveolar lavage fluid (BALF), qRT-PCR and Western blot to detect PON1 expression, lipid peroxidation and glutathione assays to quantify malondialdehyde (MDA) activity and glutathione peroxidase (GSH) levels, ELISA to determine inflammatory cytokines and immunoglobulin, and colorimetry to detect PON1 activities. Additionally, mice lung macrophages and fibroblasts were transfected with PON1 plasmid in vitro; ELISA and qRT-PCR were performed to understand the effects of PON1 on inflammatory cytokines secreted by lung macrophages, MTT assay for lung fibroblasts proliferation and qRT-PCR and Western blot for the expressions of PON1, COL1A1, and fibronectin. After overexpression of PON1, the asthma mice had decreased inflammatory cell infiltration, fibrosis degree, and airway wall thickness; inflammatory cells and inflammatory cytokines in BALF were also reduced, expressions of OVA-IgE and IgG1, and MDA activity were decreased, but the expressions of OVA-IgG2a and INF-γ and GSH levels were increased. Besides, PON1 significantly inhibited microphage expression of LPS-induced inflammatory cytokines, lung fibroblast proliferation, and COL1A1 and fibronectin expression. Thus, PON1 could relieve airway inflammation and airway remodeling in asthmatic mice and inhibit the secretion of LPS-induced macrophage inflammatory cytokines and the proliferation of lung fibroblasts.

Topics: Administration, Inhalation; Airway Remodeling; Animals; Aryldialkylphosphatase; Asthma; Bronchoalveolar Lavage Fluid; Cell Proliferation; Cells, Cultured; Cytokines; Disease Models, Animal; Female; Fibroblasts; Humans; Macrophages; Male; Mice; Ovalbumin

Asthma is one of the most common allergic diseases. Our previous studies have reported that FIP-fve in acute allergic mouse model can reduce inflammation, improve the balance of the Th1/Th2 system. However, the effects of reducing airway remodeling on FIP-fve is still unknown.. We hypothesized that orally administrated FIP-fve should be able to reduce airway remodeling in chronic allergic models.. The chronic asthma animal model was established with 6-8 weeks female Balb/c mice. After intranasal challenges with OVA, the airway inflammation and AHR were determined by a BUXCO system. BALF was analyzed with Liu's stain and ELISA assay. Lung histopathologic changes and Collagen deposition were assayed with H&E, Masson's trichrome and IHC stain.. FIP-fve significantly decreased the number of infiltrating inflammatory cells and Th2 cytokines and increased Th1 cytokines in BALF and serum compared with the OVA sensitized mice. FIP-fve had a better effect than corticosteroid could reduce infiltrating cells in lung especially neutrophils and eosinophils. We also found that the oral FIP-fve group suppressed IL-17 and enhanced IL-22 in the serum and BALF. In addition, oral FIP-fve decreased MMP9 expression, collagen expression and airway remodeling in lung tissues.. FIP-fve had anti-inflammatory effects on OVA-induced airway inflammation and an effect to inhibited Th17 cells to reduced airway remodeling and collagen expression. Moreover, FIP-fve might be a potential alternative therapy for allergic airway diseases.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Chronic Disease; Disease Models, Animal; Female; Fungal Proteins; Immunomodulation; Inflammation; Interleukin-17; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Th17 Cells; Th2 Cells

Ovalbumin (OVA) sensitization has limitations in modelling asthma. Thus, we examined the value of allergic sensitization using a purified natural allergen, house dust mite (HDM), over the sensitization performed with OVA. Mice were sham-treated, or sensitized with OVA- or HDM with identical chronology. Airway resistance, tissue damping and elastance were assessed under control conditions and after challenging the animals with methacholine (MCh) and the specific allergen. Inflammatory profile of the bronchoalveolar lavage fluid was characterized and lung histology was performed. While no difference in the lung responsiveness to the specific allergen was noted, hyperresponsiveness to MCh was observed only in the HDM-sensitized animals in the lung peripheral parameters. Lung inflammation differed between the models, but excessive bronchial smooth muscle remodelling occurred only with OVA. In conclusion, we demonstrate that a purified natural allergen offers a more relevant murine model of human allergic asthma by expressing the key features of this chronic inflammatory disease both in the lung function and structure.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Bronchoconstrictor Agents; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Female; Lung; Methacholine Chloride; Mice, Inbred BALB C; Muscle, Smooth; Ovalbumin; Pneumonia; Pyroglyphidae; Respiratory Mechanics

The root of Peucedanum japonicum Thunberg is traditionally used to treat coughs, colds, headache and inflammatory diseases in Korea and Japan. Its effects on allergic lung inflammation have not been investigated.. To investigate the anti-asthmatic effects of Peucedanum japonicum extract (PJE) using a murine model of asthma and a lipopolysaccharide (LPS)-stimulated macrophage cell line.. Mice underwent two rounds of sensitization with ovalbumin 1 week apart followed by four intranasal ovalbumin challenges on days 13-16. The control group received saline only. Two ovalbumin-sensitized groups were orally administered vehicle or PJE (200mg/kg) 5 days a week starting 1 week before the first ovalbumin sensitization. The third group was orally administered the asthma medication Montelukast (10mg/kg) on days 12-16. All animals were sacrificed on day 17. The lungs were assessed for histological features, inflammatory cell infiltration, Th2 cell activation and GATA-binding protein-3 (GATA-3) expression. The bronchoalveolar lavage fluid (BALF) was assessed for type 2 cytokine levels. The effect of PJE on the in vitro Th2 polarization of naïve CD4. PJE treatment inhibited OVA-induced inflammatory cell infiltration, eosinophilia, Th2 activation, and GATA-3 expression in the lung, reduced the interleukin (IL)-5 and IL-13 levels in BALF, down-regulated Th2 activation in vitro, and inhibited the macrophage production of inducible nitric oxide, cyclooxygenase-2, tumor necrosis factor-α, and IL-6.. PJE attenuated allergic airway inflammation by inhibiting Th2 cell activation and macrophage production of inflammatory mediators. Peucedanum japonicum may be candidate therapy for allergic lung inflammation.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Apiaceae; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cell Survival; Cytokines; Female; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; Plant Extracts; Plant Roots; RAW 264.7 Cells; Th2 Cells

It is recognized that IL-18 is related to development of asthma, but role of IL-18 in asthma remains controversial and confusing. This is largely due to lack of information on expression of IL-18 binding protein (BP) and IL-18 receptor (R) in asthma. In this study, we found that plasma levels of IL-18 and IL-18BP were elevated in asthma. The ratio between plasma concentrations of IL-18 and IL-18BP was 1:12.8 in asthma patients. We demonstrated that 13-fold more monocytes, 17.5-fold more neutrophils and 4.1-fold more B cells express IL-18BP than IL-18 in asthmatic blood, suggesting that there is excessive amount of IL-18BP to abolish actions of IL-18 in asthma. We also discovered that more IL-18R+ monocytes, neutrophils and B cells are located in asthmatic blood. Once injected, IL-18 eliminated IL-18R+ monocytes in blood, but up-regulated expression of IL-18R in lung macrophages of OVA-sensitized mice. Our data clearly indicate that the role of IL-18 in asthma is very likely to be determined by balance of IL-18/IL-18BP/IL-18R expression in inflammatory cells. Therefore, IL-18R blocking or IL-18BP activity enhancing therapies may be useful for treatment of asthma.

Topics: Adolescent; Adult; Aged; Animals; Asthma; Case-Control Studies; Female; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-18; Lipopolysaccharide Receptors; Lung; Macrophages; Male; Mice, Inbred BALB C; Middle Aged; Models, Biological; Monocytes; Neutrophils; Ovalbumin; Receptors, Interleukin-18; Young Adult

Cigarette smoke is well known to worsen asthma symptoms in asthmatic patients and to make them refractory to treatment, but the underling molecular mechanism is unclear. We hypothesized that cigarette smoke can reduce the expression of HDAC2 in asthma and the process was achieved by activating the PI3K-δ/Akt signaling pathway. We further hypothesized that roxithromycin (RXM) can alleviate the impacts by cigarette smoke.. A murine model of asthma induced by ovalbumin (OVA) and cigarette smoke has been established. The infiltration of inflammatory cells and inflammatory factors was examined in this model. Finally, we evaluated the expression of HDAC2, Akt phosphorylation levels, and the effects of RXM treatment on the model described earlier.. Cigarette smoke exposure reduced HDAC2 protein expression by enhancing the phosphorylation of Akt in PI3K-δ/Akt signaling pathway. Furthermore, RMX reduced the airway inflammation and improved the level of expression of HDAC2 in the cigarette smoke-exposed asthma mice.. This study provides a novel insight into the mechanism of cigarette smoke exposure in asthma and the effects of RXM treatment on this condition. These results may be helpful for treating refractory asthma and emphasizing the need for a smoke-free environment for asthmatic patients.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Bacterial Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Female; Histone Deacetylase 2; Lung; Mice, Inbred BALB C; Neutrophils; Nicotiana; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Roxithromycin; Smoke

Di (ethylhexyl) phthalate (DEHP) is a commonly used phthalates (PAEs) compound as plasticizer and becomes a severe environmental pollutant worldwide. Studies show that DEHP, as an environmental endocrine disruptor, has potential adverse effects on human. Epidemiologic studies indicate that DEHP is positively correlated to allergic diseases. Maternal exposure to DEHP may contribute to the increasing incidence of allergic diseases in offspring. However, the role of DEHP and its detailed mechanism in allergic disease of the offspring are still unclear. The aim of our study is to investigate whether DEHP maternal exposure could aggravate the allergic responses in offspring and its mechanism. Pregnant Wistar rats were randomly divided into three groups and exposed to different doses of DEHP. Half of the offspring were challenged with OVA after birth. All the pups of each group were sacrificed at postnatal day (PND)14, PND21 and PND28. The number of inflammatory cells in bronchoalveolar lavage was counted, lung pathological changes were observed, Th2 type cytokines expressions were checked, and the expression of TSLP signaling pathway were examined. Our results showed that maternal exposure to DEHP during pregnancy and lactation aggravated the eosinophils accumulation and the pathological inflammatory changes in pups' lung after OVA challenge. And maternal exposure to DEHP during pregnancy and lactation also elevated the levels of typical Th2 cytokines in OVA-challenged rats. What's more, maternal exposure to DEHP during pregnancy and lactation increased the levels of TSLP, TSLPR and IL-7R in the offspring after OVA challenge. Our study suggested that DEHP maternal exposure could aggravate the OVA-induced asthmatic responses in offspring. And this adjuvant effect of DEHP was related with the TSLP/TSLPR/IL-7R and its downstream signal pathways.

Topics: Animals; Asthma; Cytokines; Diethylhexyl Phthalate; Female; Inflammation; Maternal Exposure; Ovalbumin; Plasticizers; Pregnancy; Prenatal Exposure Delayed Effects; Rats; Rats, Wistar; Receptors, Interleukin-7; Serine Endopeptidases

This study aimed to explore the effect of the TSLP-DC-OX40L pathway in asthma pathogenesis and airway inflammation in mice. For this, 65 male BALF/c mice were distributed among the control, asthma, immunoglobulin G (IgG) + asthma (IgG, 500 μg/500 μL, intratracheal injection of 50 μL each time), LY294002 (OX40L inhibitor) + asthma (intratracheal injection of 2 mg/kg LY294002), and anti-TSLP + asthma (intratracheal injection of 500 μg/500 μL TSLP antibody, 50 μL each time) groups. ELISA was applied to measure the serum levels of immunoglobulin E (IgE), ovalbumin (OVA)-sIgE, interleukin-4 (IL-4), IL-5, IL-13, and interferon-γ (IFN-γ); flow cytometry was employed to detect Treg cells and dendritic cell (DC) and lymphopoiesis. RT-qPCR and Western blot assays were used to measure the levels of TSLP, OX40L, T-bet, GATA-3, NF-κB, p38, and ERK. Treatment with LY294002 and anti-TSLP resulted in increases in the numbers of total cells, eosinophils, neutrophils, and lymphocytes in the bronchoalveolar lavage fluid; total serum levels of IgE, OVA-sIgE, IL-4, IL-5, and IL-13; levels of DC cells; lymphopoiesis; and levels of TSLP, OX40L, GATA-3, NF-κB, p38, and ERK, whereas there were decreases in the levels of IFN-γ and CD4

Topics: Animals; Asthma; Chromones; Cytokines; Dendritic Cells; Disease Models, Animal; Eosinophils; Immunoglobulin E; Immunoglobulins; Inflammation; Interferon-gamma; Interleukin-13; Male; Mice; Morpholines; Ovalbumin; OX40 Ligand; Receptors, Cytokine

Asthma is a chronic airway disease characterized by airway eosinophilic inflammation and remodeling, which are associated with a loss in lung function. Although both contribute significantly to asthma pathogenesis, mechanistic studies and drug discovery have focused on inflammatory targets. In this study, we investigated the effect of the leukotriene receptor antagonist pranlukast on allergic airway inflammation and remodeling in vivo and in vitro.. Four groups of female BALB/c mice (control; ovalbumin [OVA]-sensitized and -challenged; dimethyl sulfoxide [DMSO]-treated OVA; and pranlukast-treated OVA) were examined. Lung pathology, cytokine production, and airway hyperresponsiveness (AHR) measurements were compared among these groups. A human fetal lung fibroblast HFL-1 cell line was used in the peribranchial fibrosis analysis.. OVA-sensitized and -challenged mice exhibited allergic airway inflammation and significant increases in Th2 cytokines. Pranlukast-treated mice showed significant attenuation of allergic airway inflammation. The pranlukast treatment decreased AHR and attenuated airway remodeling to goblet cell hyperplasia, collagen deposition, α-smooth muscle actin expression, and pro-fibrotic gene expression. We further demonstrated that pranlukast not only inhibited transforming growth factor-beta 1 (TGF-β1)-induced Smad signaling in human fetal lung fibroblast cells but also simultaneously reduced collagen synthesis and pro-fibrotic gene expression.. The leukotriene receptor antagonist pranlukast can reduce airway inflammation and remodeling by inhibiting TGF-β/Smad signaling in an OVA-sensitized and -challenged asthma mouse model, thus suppressing AHR.

Topics: Airway Remodeling; Animals; Asthma; Cell Line; Chromones; Collagen; Disease Models, Animal; Female; Fibroblasts; Humans; Leukotriene Antagonists; Mice; Mice, Inbred BALB C; Ovalbumin; Signal Transduction; Smad Proteins; Transforming Growth Factor beta1

Topics: Adoptive Transfer; Animals; Asthma; Calcium-Binding Proteins; Cell Differentiation; Cell Proliferation; Dendritic Cells; Down-Regulation; Female; Hypersensitivity; Immunoglobulin E; Intercellular Signaling Peptides and Proteins; Interleukin-12; Lymphocyte Activation; Mice; Ovalbumin; Th1 Cells; Th2 Cells

Cathelicidin has been reported to be multifunctional. The current study aimed to investigate the influences of exogenous cathelicidin-related antimicrobial peptide (CRAMP) on inflammatory responses in different disease models. In OVA-induced allergic airway inflammation, CRAMP significantly enhanced the infiltration of inflammatory cells and accumulation of proinflammatory Th2 cytokine IL-13 and IL-33 in bronchial alveolar lavage fluid (BALF), exacerbated lung tissue inflammation and airway goblet cell hyperplasia, and elevated OVA-specific IgE level in serum. In oxazolone-induced intestinal colitis, the expression levels of CRAMP and its receptor FPR2 significantly increased in comparison with those of TNBS-induced mice, vesicle and normal controls. Exogenous CRAMP significantly prevented the development of ulcerative colitis, evidenced by improved body weight regain, decreased colons weight/length ratio, elevated epithelial integrity, and ameliorated colon tissue inflammation. In addition, pro-inflammatory cytokines TNF-α, IL-1β, IL-4 and IL-13, as well as chemokines CXCL2 and CXCL5 for neutrophils recruitment were significantly decreased in CRAMP-treated mice, and epithelial repair-related factors MUC2 and Claudin1 were increased, determined by real time-PCR and ELISAs. The results indicated that although CRAMP has pro-inflammatory effects in airway, local application of exogenous CRAMP might be a potential approach for the treatment of ulcerative colitis.

Topics: Administration, Intranasal; Administration, Rectal; Animals; Antimicrobial Cationic Peptides; Asthma; Bronchoalveolar Lavage Fluid; Cathelicidins; Colitis, Ulcerative; Cytokines; Disease Models, Animal; Disease Progression; Female; Goblet Cells; Humans; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Oxazolone

Asthma is a long term inflammatory disease of the airway of lungs characterized by variable airflow obstruction and bronchospasm. Asthma is caused by a complex combination of environmental and genetic interactions. In this study, we conducted proteomic analysis of samples derived from control and OVA challenged mice for environmental respiratory disease by using 2-D gel electrophoresis. In addition, we explored the genes associated with the environmental substances that cause respiratory disease and conducted RNA-seq by next-generation sequencing. Proteomic analysis revealed 7 up-regulated (keratin KB40, CRP, HSP27, chaperonin containing TCP-1, TCP-10, keratin, and albumin) and 3 down-regulated proteins (PLC-α, PLA2, and precursor ApoA-1). The expression diversity of many genes was found in the lung tissue of OVA challenged moue by RNA-seq. 146 genes were identified as significantly differentially expressed by OVA treatment, and 118 genes of the 146 differentially expressed genes were up-regulated and 28 genes were downregulated. These genes were related to inflammation, mucin production, and airway remodeling. The results presented herein enable diagnosis and the identification of quantitative markers to monitor the progression of environmental respiratory disease using proteomics and genomic approaches.

Topics: Animals; Asthma; Female; Inflammation; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; Proteomics; Transcriptome

Vitexin is an important component of various medicinal plants frequently used to treat asthma, such as Crataegus spp., Vitex spp., Passiflora spp., and Echinodorus spp. However, there is no information about the vitexin potential as anti-asthmatic. The aim of the present work was to evaluate the anti-hypersensitive activity of vitexin in a murine ovalbumin (OVA)-induced allergic asthma model. Mice were sensitized to OVA by i.p. injection on days 1st and 10th, followed by a daily challenge with OVA using a nebulizer, from days 19th to 24th. Vitexin or dexamethasone were orally administered 1h before each OVA challenge. Vitexin attenuates migration induced by OVA-hypersensitivity of eosinophil, neutrophil, and mononuclear cells in bronchoalveolar lavage fluid (BALF). Histological analysis of the lungs shows that vitexin suppressed leukocyte infiltration, mucus production and pulmonary edema. Increases in Th2 cytokines in BALF in OVA-induced asthma is also attenuated by vitexin, as well as plasma levels of IgE. Overall, these results suggest that vitexin can suppress OVA-induced allergic inflammation in mice and provide a strong rationale for further developing vitexin as a candidate treatment for allergic hypersensitivity. These data corroborate the popular use of vitexin-rich plants for asthma treatment.

Topics: Animals; Anti-Asthmatic Agents; Apigenin; Asthma; Bronchoalveolar Lavage Fluid; Dose-Response Relationship, Drug; Inflammation; Inflammation Mediators; Male; Mice; Ovalbumin

Asthma is a chronic respiratory disease related to hyper‑responsiveness. The majority of patients suffer mild symptoms, however, some cases, especially in the young and the elderly, can lead to death by apnea. Mycoleptodonoides atichisonii (M. atichisonii) is an edible mushroom that has previously been reported to possess several bioactive properties, such as the synthesis of nerve growth factors, anti‑obesity effects and the ability to prevent cell death. In the current study, the authors evaluated the anti‑asthmatic effects of M. atichisonii using an ovalbumin‑induced asthma mouse model. M. atichisonii dose‑dependently suppressed the levels of white blood cells, eosinophils and immunoglobulin (Ig)E in BALB/c mice, resulting from ovalbumin‑induced asthma. M. atichisonii recovered the typical asthmatic morphological changes in lungs, such as mucous hyper‑secretion, epithelial layer hyperplasia, eosinophil infiltration and various cell surface molecules, such as CD3, CD4, CD8, CD19 and major histocompatibility complex class II. With the exception of CD19+ cells and IL‑12p40, M. atichisonii affected almost all factors related to asthma induction including the T helper (Th)1/Th2 transcription factors, T‑bet and GATA‑3, Th1‑related cytokines, Th2‑related cytokines and proinflammatory cytokines. In addition, M. atichisonii significantly inhibited the expression of IL‑5, IL‑13 and IL‑6. The authors concluded that M. atichisonii may be a promising drug candidate against asthma.

Topics: Agaricales; Animals; Anti-Asthmatic Agents; Asthma; Biological Products; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Eosinophils; Female; GATA3 Transcription Factor; Gene Expression Regulation; Histocompatibility Antigens Class II; Humans; Immunomodulation; Leukocytes; Mice; Niacin; Oleic Acid; Ovalbumin; T-Box Domain Proteins; Th1 Cells; Th2 Cells

The hygiene hypothesis is the leading concept to explain the current asthma epidemic, which is built on the observation that a lack of bacterial contact early in life induces allergic T. Because little is known about the contribution of respiratory tract viruses in this context, we evaluated the effect of prior influenza infection on the development of allergic asthma.. Mice were infected with influenza and, once recovered, subjected to an ovalbumin- or house dust mite-induced experimental asthma protocol. Influenza-polarized effector memory T (Tem) cells were transferred adoptively to allergen-sensitized animals before allergen challenge. A comprehensive in silico analysis assessed homologies between virus- and allergen-derived proteins. Influenza-polarized Tem cells were stimulated ex vivo with candidate peptides. Mice were immunized with a pool of virus-derived T-cell epitopes.. In 2 murine models we found a long-lasting preventive effect against experimental asthma features. Protection could be attributed about equally to CD4. For the first time, our results illustrate heterologous immunity of virus-infected animals toward allergens. This finding extends the original hygiene hypothesis.

Topics: Allergens; Animals; Asthma; Epitopes, T-Lymphocyte; Female; Influenza A Virus, H1N1 Subtype; Mice, Inbred BALB C; Orthomyxoviridae Infections; Ovalbumin; Peptides; Pyroglyphidae; T-Lymphocytes

KIF3A expression was decreased in asthmatic child patients and animal. Impaired KIF3A expression resulted in increased Th2 inflammation in mice and apoptosis in renal tubular epithelium and photoreceptor cells. This work aimed to investigate the role of KIF3A in epithelium apoptosis and bronchial inflammation in asthma.. After establishment of ovalbumin induced asthma, the mice were infected with KIF3A adenovirus through nasal cavity inhalation. KIF3A expression and apoptosis in epithelia of nasal mucosa and bronchia were determined using qRT-PCR, western blotting, immunohistochemistry and TUNEL staining. The mRNA expression of COX-2, IL-4, IL-5, IL-13, IL-6, IL-10 and TNF-α was also measured. In vitro, human bronchial epithelial cell line 16HBE 14o- was stimulated with IL-4, IL-13 and TNF-α, accompanied by KIF3A knockdown or overexpression using siRNA or KIF3A adenovirus respectively. Apoptosis, mRNA expression of CCL17, CCL26, IL-5 and IL-8, and protein expression of COX-2 and β-catenin were determined using flow cytometry, qRT-PCR and western blotting.. KIF3A expression was reduced in epithelia of nasal mucosa and bronchia of asthmatic mice, and overexpression of KIF3A ameliorated epithelial cell apoptosis and bronchial inflammation in asthmatic mice. In vitro, KIF3A knockdown significantly promoted epithelium apoptosis, facilitated the transcription of CCL17, CCL26, IL-5 and IL-8, and increased the protein levels of COX-2 and β-catenin translocation, whereas overexpression of KIF3A exhibited the opposite effect.. KIF3A plays an important role in epithelium apoptosis and bronchial inflammation in asthma, and may be a potential target for asthma treatment.

Topics: Adenoviridae; Animals; Apoptosis; Asthma; Bronchi; Female; Flow Cytometry; Gene Expression Regulation; Gene Knockdown Techniques; Humans; In Situ Nick-End Labeling; Inflammation; Kinesins; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Mucosa; RNA, Messenger

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Fatty Acids; Female; Humans; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; Sea Cucumbers; Th17 Cells; Th2 Cells

A series of novel α-terpineol derivatives were designed and synthesized through structural derivatization of the tertiary hydroxyl moiety or reduction of the double bond. Of the resulting compounds, eight compounds enhanced relaxation of airway smooth muscle (ASM) compared to the α-terpineol precursor, and four compounds (4a, 4d, 4e, and 4i)were superior or comparable to aminophylline at a concentration of 0.75 mmol/L. Assays for 3'-5'-Cyclic adenosine monophpsphate (cAMP) activation revealed that some representative α-terpineol derivatives in this series were capable of upregulating the level of cAMP in ASM cells. Further in vivo investigation using the asthmatic rat model, illustrated that treatment with the compounds 4a and 4e resulted in significantly lowered lung resistance (RL) and enhanced dynamic lung compliance (Cldyn), two important parameters for lung fuction. Moreover, treatment with 4e downregulated the levels of both IL-4 and IL-17. Due to its several favorable physiological functions, including ASM relaxation activity, cAMP activation capability, and in vivo anti-asthmatic efficacy, 4e is a promising remedy for bronchial asthma, meriting extensive development.

Topics: Administration, Oral; Aluminum Hydroxide; Animals; Anti-Asthmatic Agents; Asthma; Cyclohexane Monoterpenes; Cyclohexenes; Dose-Response Relationship, Drug; Drug Discovery; Guinea Pigs; Injections, Intraperitoneal; Male; Molecular Structure; Monoterpenes; Ovalbumin; Rats; Rats, Sprague-Dawley; Structure-Activity Relationship

Asthma is a chronic disease associated with hyperresponsiveness, obstruction and remodeling of the airways. Epithelial-mesenchymal transition (EMT) has an important role in these alterations and may account for the accumulation of subepithelial mesenchymal cells, thus contributing to airway hyperresponsiveness and remodeling. Epigallo-catechin‑3‑gallate (EGCG), which is a type of polyphenol, is the most potent ingredient in green tea, and exhibits antibacterial, antiviral, antioxidative, anticancer and chemopreventive activities. Recently, numerous studies have investigated the protective effects of EGCG against asthma and other lung diseases. In the present study, the role of EGCG in ovalbumin (OVA)‑challenged asthmatic mice was determined. In addition, the inhibitory effects of EGCG against transforming growth factor (TGF)‑β1‑induced EMT and migration of 16HBE cells, and the underlying mechanisms of the phosphatidylinositol 3‑kinase/protein kinase B (PI3K/Akt) signaling pathway, were investigated by immunofluorescence, Transwell, wound healing assay and western blot analysis, respectively. The results indicated that EGCG may suppress inflammation and inflammatory cell infiltration into the lungs of OVA‑challenged asthmatic mice, and may also inhibit EMT via the PI3K/Akt signaling pathway through upregulating the expression of phosphatase and tensin homolog (PTEN) in vivo and in vitro. The present study also revealed the anti‑migratory effects of EGCG in TGF‑β1‑induced 16HBE cells, thus suggesting it may reduce airway remodeling. The present study provides a novel insight into understanding the protective effects of EGCG on airway remodeling in asthma, and indicates that EGCG may be useful as an adjuvant therapy for bronchial asthma.

Topics: Airway Remodeling; Animals; Asthma; Catechin; Disease Models, Animal; Epithelial-Mesenchymal Transition; Humans; Inflammation; Mice; Oncogene Protein v-akt; Ovalbumin; Phosphatidylinositol 3-Kinases; PTEN Phosphohydrolase; Signal Transduction; Tea

Allergic asthma during early life period has been reported to be associated with neurochemical and behavioral disorders, including anxiety and depression. We aimed to determine the effects of exercise on depressive- and anxiety-like behaviors as well as lung and hippocampus oxidative stress in ovalbumin (OVA)-sensitized juvenile rats. Animals were divided into 4 groups including control (non-exercised and non-sensitized), Exe (exercise and non-sensitized); OVA (non-exercised and OVA-sensitized); and OVA+Exe (exercised and OVA-sensitized). The rats were subjected to chronic OVA sensitization followed by 4 weeks of treadmill exercise training. Compared to the control group, the OVA group had an increase in anxiety- and depressive-like behavior, lung inflammation, and oxidative stress index in the lung and hippocampus. Compared to the OVA group, the OVA+Exe group had a decline in anxiety- and depressive-like behavior, lung inflammation, and oxidative stress index in the lung and hippocampus. No significant difference in terms of the above-mentioned parameters, were found between the control group and the Exe group. Exercise decreased depressive- and anxiety-like behaviors in OVA-sensitized juvenile rats; this effect might have been mainly mediated by improvement in antioxidant system.

Topics: Allergens; Animals; Anxiety; Asthma; Bronchoalveolar Lavage Fluid; Depression; Disease Models, Animal; Hippocampus; Male; Malondialdehyde; Maze Learning; Ovalbumin; Oxidative Stress; Physical Conditioning, Animal; Rats; Rats, Wistar; Sulfhydryl Compounds; Superoxide Dismutase; Swimming

V-domain immunoglobulin suppressor of T-cell activation (VISTA) is a novel immune checkpoint receptor and ligand that regulates T-cell activation. We investigated the functional involvement of VISTA in Th2 cell-mediated immune responses using an ovalbumin (OVA)-induced allergic asthma model. Treatment with an anti-VISTA monoclonal antibody (mAb) during allergen sensitization increased the production of antibodies, including total IgE, OVA-specific IgG1 and IgG2a and allergen-specific IL-5 and IL-13; it also increased the expression of IL-13 by splenic CD4+ T cells. However, treatment with the anti-VISTA mAb during sensitization did not accelerate asthmatic responses, including airway hyper-responsiveness (AHR) or the number of eosinophils in bronchoalveolar lavage (BAL) fluid. In contrast, treatment with the anti-VISTA mAb during allergen challenge significantly augmented AHR and BAL fluid eosinophilia. This treatment also increased the production of IL-5 and IL-13 in BAL fluid and the expression of IL-13 by CD4+ T cells in draining lymph nodes. These results suggest that VISTA is involved in the regulation of Th2 cell generation and Th2 cell-mediated antibody production and regulates asthmatic responses, especially in the effector phase.

Topics: Allergens; Animals; Antibodies, Monoclonal; Antigen-Antibody Reactions; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; CD4-Positive T-Lymphocytes; Disease Models, Animal; Eosinophils; Female; Flow Cytometry; Membrane Proteins; Mice; Mice, Inbred BALB C; Ovalbumin; Th2 Cells

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchi; Bronchoalveolar Lavage Fluid; Cytokines; Fenofibrate; Immunoglobulin E; Leukocyte Count; Lipopolysaccharides; Lung; Male; Ovalbumin; PPAR alpha; Rats, Wistar

IL-10-differentiated dendritic cells (DC10) can reverse the asthma phenotype in mice, but how they suppress the asthmatic B cell response is unclear. Herein we assessed the mechanism(s) by which DC10 and DC10-induced Treg affect IgG1 production in asthma. We observed a rapid decline in lung-resident OVA-specific IgG1-secreting B cells on cessation of airway allergen challenge, and intraperitoneal DC10 therapy did not amplify that (p>0.05). It did however increase the loss of IgG1-B cells from the bone marrow (by 45+/-7.2%; p≤0.01) and spleen (by 65+/-17.8%; p≤0.05) over 2 wk. Delivery of OVA-loaded DC10 directly into the airways of asthmatic mice decreased the lung IgG1 B cell response assessed 2 dy later by 33+/-9.7% (p≤0.01), while their co-culture with asthmatic lung cell suspensions reduced the numbers of IgG1-secreting cells by 56.5+/-9.7% (p≤0.01). This effect was dependent on the DC10 carrying intact allergen on their cell surface; DC10 that had phagocytosed and fully processed their allergen were unable to suppress B cell responses, although they did suppress asthmatic Th2 cell responses. We had shown that therapeutic delivery of DC10-induced Treg can effectively suppress asthmatic T and B cell (IgE and IgG1) responses; herein CD4+ cells or Treg from the lungs of DC10-treated OVA-asthmatic mice suppressed in vitro B cell IgG1 production by 52.2+/-8.7% (p≤0.001) or 44.6+/-12.2% (p≤0.05), respectively, but delivery of DC10-induced Treg directly into the airways of asthmatic mice had no discernible impact over 2 dy on the numbers of lung IgG1-secreting cells (p≥0.05). In summary, DC10 treatment down-regulates OVA-specific B cell responses of asthmatic mice. While DC10 that carry intact allergen on their cell surface can dampen this response, DC10-induced Treg are critical for full realization of this outcome. This suggests that infectious tolerance is an essential element in regulatory DC control of the B cell response in allergic asthma.

Topics: Allergens; Animals; Asthma; Cell Separation; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Immunoglobulin G; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory

Annexin-A1 (ANXA1) is a glucocorticoid-induced protein with multiple actions in the regulation of inflammatory cell activation. The anti-inflammatory protein ANXA1 and its N-formyl peptide receptor 2 (FPR2) have protective effects on organ fibrosis. However, the exact role of ANXA1 in asthma remains to be determined. The aim of this study was to identify the role of ANXA1 in bronchial asthma.. In mice sensitized and challenged with ovalbumin (OVA-OVA mice) and mice sensitized with saline and challenged with air (control mice), we investigated the potential links between ANXA1 levels and bronchial asthma using ELISA, immunoblotting, and immunohistochemical staining. Moreover, we also determined ANXA1 levels in blood from 50 asthmatic patients (stable and exacerbated states).. These results suggest that ANXA1 may be a potential marker and therapeutic target for asthma.

Topics: Adult; Aged; Animals; Annexin A1; Asthma; Bronchoalveolar Lavage Fluid; Case-Control Studies; Female; Forced Expiratory Volume; Humans; Lung; Male; Mice; Mice, Inbred BALB C; Middle Aged; Ovalbumin; Symptom Flare Up; Vital Capacity

•Ad5-gsgAM elicits Th1 responses and suppresses Th2-mediated allergic asthma in mice. •Ad5-gsgAM inhibits IL-33/ST2 axis by reducing IL-33 secretion but not ILC2 recruiting. •Treg is essential for modulating Th2 responses and IL-33/ST2 axis by Ad5-gsgAM.

Topics: Adenoviridae; Animals; Antigens, Bacterial; Asthma; Cytokines; Female; Genetic Vectors; Immunization; Immunoglobulin E; Mice, Inbred C57BL; Mycobacterium tuberculosis; Ovalbumin; Th1 Cells; Th2 Cells; Tuberculosis Vaccines

An experimental model of ovalbumin (OVA) induced asthma was used to assess the effects of nebivolol, the third-generation selective β

Topics: Animals; Anti-Asthmatic Agents; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Gene Expression Regulation; Gene Expression Regulation, Enzymologic; Guinea Pigs; Histamine; Interleukin-6; Leukocyte Count; Lung; Male; Malondialdehyde; Nebivolol; Nitric Oxide Synthase Type II; Nitric Oxide Synthase Type III; Ovalbumin; Oxidative Stress; Respiration; Tumor Necrosis Factor-alpha

The transcription factor nuclear factor (NF)-κB plays a pivotal role in the development of allergic airway inflammation. However, the mechanism of NF-κB activation in asthma remains to be elucidated.. CK2α activation was assessed by CK2α phosphorylation and protein expression. Airway levels of histamine and cytokines were determined by ELISA. We used 2 (active and passive) forms of allergic pulmonary inflammation models. In the active form, the animals were immunized with ovalbumin (OVA) intraperitoneally, followed by an airway challenge with OVA. In the passive form, the animals were passively sensitized by intratracheal instillation with either anti-OVA IgE or anti-OVA IgG, followed by an airway challenge with OVA. The role of NADPH oxidase (NOX) in CK2α activation was assessed using NOX2-/- and NOX4-/- mice because NOX2 and NOX4 contribute to many inflammatory diseases.. The second airway challenge increased CK2α phosphorylation and protein expression in airway epithelial cells as well as nuclear translocation of the p50 and p65 subunits of NF-κB, all of which were inhibited by the CK2α inhibitor 4,5,6,7-tetrabromobenzotriazole and the antioxidant N-acetyl-L-cysteine. CK2α phosphorylation and protein expression were significantly impaired in NOX2-/-, but not in NOX4-/-, mice. Induction of passive sensitization using anti-OVA IgE activated neither CK2α nor NF-κB. In contrast, induction of passive sensitization using anti-OVA IgG activated both CK2α and NF-κB.. These data suggest that Fcγ receptor/reactive oxygen species/CK2α is a key inducer of NF-κB activation in airway epithelial cells in a murine model of asthma.

Topics: Allergens; Animals; Asthma; Casein Kinase II; Cytokines; Disease Models, Animal; Female; Histamine; Humans; Immunization; Immunoglobulin E; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; NADPH Oxidase 2; NADPH Oxidase 4; NF-kappa B; Ovalbumin; Phosphorylation; Reactive Oxygen Species; Receptors, IgG

Notch1 has been linked to the pathogenesis of asthma due to its contribution on Th1/Th2 imbalance. γ-Secretase inhibitor (GSI) acts as an effective blocker of Notch1 signaling. Glucocorticoids (GCs) are the most effective anti-inflammatory drugs for asthma. The present study investigated the involvement of the Notch1 pathway in the anti-inflammatory effect of GCs and its association with Th1/Th2 balance.. The asthma model was established in BALB/c mice by sensitization with ovalbumin (OVA). Dexamethasone (DEX; 1 mg/kg) and/or GSI (0.03 mg/kg) was orally or intranasally administrated.. Compared to the OVA-sensitized mice, the administration of DEX and/or GSI significantly ameliorated the airway inflammation infiltration, goblet cell metaplasia, and airway hyper-responsiveness. The expression of IL-4 and IL-13, as well as the ratios of eosinophils and lymphocytes, were significantly decreased, whereas IFN-γ and IL-2 levels were significantly increased in bronchoalveolar lavage fluid after the administration of DEX and GSI. The expressions of the Notch1/NICD1 pathway were decreased after DEX and/or GSI administration in lung tissues, especially in CD4+ T cells. Also, a reduction of GATA3 and elevation of T-bet levels were correlated with the upregulation of Th1/Th2 ratios in lung tissues.. Through the inhibition of Notch1 signaling, both GSI and GCs could regulate Th1/Th2 balance involved in allergic airway inflammation in OVA-induced asthma.

Topics: Allergens; Animals; Asthma; Cells, Cultured; Cytokines; Dexamethasone; Disease Models, Animal; Glucocorticoids; Goblet Cells; Humans; Metaplasia; Mice; Mice, Inbred BALB C; Ovalbumin; Receptor, Notch1; Signal Transduction; T-Box Domain Proteins; Th1-Th2 Balance

The aim of the present study was to explore the roles of OX40/OX40 ligand (OX40L) signaling and OX40+ T cells in ovalbumin (OVA)‑induced mouse asthma model. Asthma was induced by OVA exposure and subsequent co‑treatment with OX40L protein, neutralizing anti‑OX40L blocking antibody, OX40+ T cells or PBS. The protein expression levels of interleukin (IL)‑4, IL‑6, IL‑13, IL‑17, tumor necrosis factor (TNF)‑α and interferon (IFN)‑γ in bronchoalveolar lavage fluid (BALF) were examined using murine cytokine‑specific ELISA. Eosinophil accumulation as well as proliferation and apoptosis of T cells in BALF were detected by Cell Counting kit‑8 and flow cytometric assays. Expression of the apoptosis‑related protein cleaved caspase‑3 was examined in OX40+ T cells using western blot assay. Flow cytometric analysis revealed that OVA‑treated mice that were co‑treated with OX40L or OX40+ T cells exhibited higher eosinophil infiltration compared with control mice treated only with OVA, whereas neutralizing anti‑OX40L blocking antibody inhibited eosinophil infiltration. ELISA assays demonstrated that the expression of IL‑4, IL‑6, IL‑13, IL‑17, TNF‑α and IFN‑γ in BALF in OX40L‑treated and OX40+ T cell‑treated mice was increased compared with expression levels in control mice. Treatment with OX40L protein effectively reduced apoptosis of T cells and the expression of cleaved caspase‑3 in T cells. OX40L‑treated and OX40+ T cell‑treated mice exhibited increased asthma through OX40/OX40L signaling, which probably promoted inflammatory factor expression, eosinophil infiltration and T cell proliferation.

Topics: Animals; Antibodies, Neutralizing; Antigens, Differentiation; Apoptosis; Asthma; Bronchoalveolar Lavage Fluid; Caspase 3; CD4-Positive T-Lymphocytes; Cell Proliferation; Disease Models, Animal; Eosinophils; Female; Gene Expression Regulation; Interferon-gamma; Interleukin-13; Interleukin-17; Interleukin-4; Interleukin-6; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Ovalbumin; OX40 Ligand; Signal Transduction; Tumor Necrosis Factor Inhibitors; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors

The allergic asthma model induced by ovalbumin (OVA) was established in the immature rat. Dexamethasone (DXM) was adopted for intervention to analyze the treatment effect and to explore the relationship with toll-like receptor 4 (TLR4).. Immature SD rat was treated by OVA to construct allergic asthma model and intervened by DXM. The rats were randomly divided into model group, experimental group, and control group. The changes in lung tissue were observed by light microscope. The EOS infiltration and reactivity of airway wall were compared. The expressions of TLR2 and TLR4 protein and mRNA in the lung tissue were tested by Western blot and RT-PCR.. The lung tissue in the model group was infiltrated by a lot of inflammatory cells, and mucous membrane edema was observed, compared with that in the control group. There were only a few inflammatory cells in the interstitial tissue and pulmonary alveoli in the experimental group compared with that in the model group. EOS count of airway wall and airway reactivity decreased in the experimental group. The levels of TLR2 and TLR4 were significantly elevated in the third week compared with the first week (p<0.05).. The treatment of DXM can alleviate the pathological changes of the lung tissue in SD immature rat with allergic asthma, reduce EOS infiltration in the airway wall, decrease airway reactivity, and elevate expressions of TLR2 and TLR4.

Topics: Animals; Asthma; Dexamethasone; Disease Models, Animal; Eosinophils; Female; Gene Expression; Lung; Ovalbumin; Rats; Rats, Sprague-Dawley; Toll-Like Receptor 2; Toll-Like Receptor 4

Asthma and chronic obstructive pulmonary disease (COPD), chronic airway inflammatory diseases characterized by airflow limitation, have different etiologies and pathophysiologies. Asthma-COPD Overlap (ACO) has recently been used for patients with mixed asthma and COPD. The pathophysiological mechanisms of ACO have not been clearly understood due to the lack of an appropriate murine model. To investigate its pathophysiology, we examined a murine model by allergen challenge in surfactant protein-D (SP-D)-deficient mice that spontaneously developed pulmonary emphysema. SP-D-deficient mice were sensitized and challenged by ovalbumin (OVA). Lungs and bronchoalveolar lavage fluid (BALF) were collected for analysis, and static lung compliance and airway hyperresponsiveness (AHR) were measured 48 h after the last OVA challenge. In SP-D-deficient, naïve, or OVA-challenged mice, the mean linear intercept and static lung compliance were increased compared with wild-type (WT) mice. There was no significant difference in goblet cell hyperplasia and the gene expression of Mucin 5AC (MUC5AC) between SP-D-deficient and WT OVA-challenged mice. In SP-D-deficient OVA-challenged mice, airway hyperresponsiveness was significantly enhanced despite the lower eosinophil count and the concentration of interleukin (IL)-5 and IL-13 in BALF compared with WT OVA-challenged mice at 120 ventilations per minute. When mice were ventilated at a lower ventilation frequency of 100 ventilations per minute, elevated airway hyperresponsiveness in SP-D-deficient OVA-challenged mice was diminished. This model of emphysematous change with allergic airway inflammation raises the possibility that frequency-dependent airway hyperresponsiveness may be involved in the pathophysiology of ACO.

Topics: Animals; Asthma; Disease Models, Animal; Hypersensitivity; Inflammation; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Pulmonary Disease, Chronic Obstructive; Pulmonary Emphysema; Pulmonary Surfactant-Associated Protein D; Respiratory Hypersensitivity

Specific immunotherapy, including agonists for Toll-like receptor 2 (TLR2), have been shown to protect from allergies and to have a high immunomodulatory capacity.. A new antibody, TSP-2, reactive against an epitope of the extracellular domain of TLR2, was identified. The effect of the antibody on dendritic cells was assessed by immunohistochemistry, Western blot, and flow cytometric analysis. The effect of TSP-2 in a murine asthma model induced with ovalbumin (OVA) was assessed. The model is a form of airway hyperresponsiveness (AHR) and was analyzed by whole-body plethysmography, the measurement of Th1/Th2 cytokines in bronchial alveolar lavage fluid (BALF) and serum by ELISA, and the CCK-8 assay for lymphocyte proliferation. The effect of TSP-2 on the maturation of bone marrow-derived dendritic cells (BMDCs) was assessed by flow cytometric analysis.. TSP-2 promoted the maturation of dendritic cells and the proliferation of lymphocyte in vitro and in vivo. The effect of TSP-2 on T helper 1 (Th1)/Th2 cytokine secretion was slightly more powerful than that of Pam3CSK4. TSP-2 antibody reduced AHR and OVA-specific IgE levels in allergic asthma. TSP-2 antibody also reduced lung inflammation and decreased leukocyte numbers in an OVA-sensitized and challenged asthma model. TSP-2 antibody increased OVA-stimulated I-A, CD80, CD86, and MHC-II levels on BMDCs.. This study identifies a novel therapeutic strategy for AHR, which uses antibodies reactive against TLR2. It also provides theoretical evidence for the control of allergic asthma by targeting TLR2.

Topics: Animals; Antibodies; Asthma; Blotting, Western; Cell Line; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Immunohistochemistry; Mice; Mice, Inbred BALB C; Ovalbumin; Th1-Th2 Balance; Thrombospondins; Toll-Like Receptor 2; Treatment Outcome

Bombesin receptor subtype-3 (BRS-3) is a member of the bombesin-like peptide receptor family. Our previous studies demonstrated that activation of the human BRS-3 plays a protective role in oxidation-injured human bronchial epithelial cells (HBEC). The present study was designed to determine the role of BRS-3 activation in the antigen-presenting action of HBEC and the corresponding proliferation and differentiation of T cells.. In vivo, an asthma animal model was established and the expression and distribution of BRS-3 were analyzed by immunocytochemistry. In vitro, 2 kinds of B7 costimulatory molecules, i.e., B7H1 and B7DC, on HBEC were analyzed by flow cytometry. The antigen uptake by HBEC was examined by confocal microscopy and flow cytometry. The antigen-presenting-action-induced proliferation of T cells was determined by MTT assays. IFN-γ and IL-4 levels were measured by ELISA. All studies were performed in the absence or presence of the synthetic peptide P3513.. BRS-3 expression was induced in asthma animal models and mainly distributed in bronchial epithelial cells. HBEC express the costimulatory molecules B7H1 and B7DC. BRS-3 activation increased B7H1 expression but decreased B7DC expression on HBEC. BRS-3 activation also increased the antigen uptake by HBEC and the subsequent T cell proliferation. In addition, BRS-3 activation promoted the releases of IFN-γ, but not IL-4, in the supernatant of cocultured HBEC and T cells.. These data suggest that HBEC can present antigen to T cells and BRS-3 activation promotes the process of antigen presentation and subsequent T cell proliferation and Th1 differentiation.

Topics: Allergens; Animals; Antigen Presentation; Asthma; B7-H1 Antigen; Bronchi; Cell Differentiation; Cell Proliferation; Cells, Cultured; Disease Models, Animal; Epithelial Cells; Humans; Interferon-gamma; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Bombesin; Th1 Cells

Uncaria tomentosa (Willd. Ex Schult) DC is used by indigenous tribes in the Amazonian region of Central and South America to treat inflammation, allergies and asthma. The therapeutic properties of U. tomentosa have been attributed to the presence of tetracyclic and pentacyclic oxindole alkaloids and to phenolic acids.. To characterize aqueous bark extracts (ABE) and aqueous leaf extracts (ALE) of U. tomentosa and to compare their anti-inflammatory effects.. Constituents of the extracts were identified by ultra performance liquid chromatography-mass spectrometry. Anti-inflammatory activities were assessed in vitro by exposing lipopolysaccharide-stimulated macrophage cells (RAW264.7-Luc) to ABE, ALE and standard mitraphylline. In vivo assays were performed using a murine model of ovalbumin (OVA)-induced asthma. OVA-sensitized animals were treated with ABE or ALE while controls received dexamethasone or saline solution. Bronchial hyperresponsiveness, production of Th1 and Th2 cytokines, total and differential counts of inflammatory cells in the bronchoalveolar lavage (BAL) and lung tissue were determined.. Mitraphylline, isomitraphylline, chlorogenic acid and quinic acid were detected in both extracts, while isorhyncophylline and rutin were detected only in ALE. ABE, ALE and mitraphylline inhibited the transcription of nuclear factor kappa-B in cell cultures, ALE and mitraphylline reduced the production of interleukin (IL)-6, and mitraphylline reduced production of tumor necrosis factor-alpha. Treatment with ABE and ALE at 50 and 200 mg kg. The results clarify for the first time the anti-inflammatory activity of U. tomentosa in a murine model of asthma. Although ABE and ALE exhibited distinct chemical compositions, both extracts inhibited the production of pro-inflammatory cytokines in vitro. In vivo assays revealed that ABE was more effective in treating asthmatic inflammation while ALE was more successful in controlling respiratory mechanics. Both extracts may have promising applications in the phytotherapy of allergic asthma.

Topics: Acids, Carbocyclic; Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Cat's Claw; Cell Survival; Cytokines; Disease Models, Animal; Indole Alkaloids; Lung; Mice; Ovalbumin; Phytotherapy; Plant Bark; Plant Extracts; Plant Leaves; RAW 264.7 Cells

Diisodecyl phthalate (DIDP) is considered to be one of the less toxic phthalates. However epidemiological studies suggest that DIDP is associated with the occurrence of asthma. The effect of DIDP exposure on allergic asthma and the underlying mechanism have not been fully elucidated. Here, mice were exposed to DIDP and sensitization with OVA. The results demonstrated that DIDP exposure aggravated allergic asthma. Exposure to 15 mg/kg/day DIDP markedly exacerbated airway remodeling and promoted airway hyperresponsiveness (AhR). The study suggests that exposure to DIDP not only promotes a predominant Th2 response, but also induces Th17-type immunity. The induced allergic asthma was accompanied by elevation of IgE, an increase in TSLP expression and exacerbation of oxidative stress. Inhibition of oxidative stress by Vitamin E effectively alleviated the airway remodeling and AhR induced by DIDP and OVA sensitization. Treatment with Vitamin E inhibited the Th2 response and the production of TSLP. Blocking the activation of p38 MAPK by SB203580 prevented elevation of IL-1β and IL-17A induced by DIDP and OVA sensitization and effectively alleviated Th17 type asthmatic lesions. These results suggest that exposure to DIDP exacerbates the Th2 and Th17 response through aggravating oxidative stress and activation of the p38 MAPK pathway.

Topics: Animals; Asthma; Humans; Immunoglobulin E; Interleukin-17; Interleukin-1beta; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; p38 Mitogen-Activated Protein Kinases; Phthalic Acids; Plasticizers; Th17 Cells; Th2 Cells

The aim of this study was to explore the role of JAK/STAT signaling pathway inhibitor Ruxolitinib in neutrophilic airway inflammation and its possible immunological mechanism.. A total of 60 female C57BL/6 mice were randomly divided into neutrophilic asthma (NA) group, Ruxolitinib-treated (Ruxo) group and control (Con) group. Mice in NA and Ruxo groups were sensitized with ovalbumin (OVA) and excited to establish mice models of asthma. Bronchoalveolar lavage fluid (BALF) was collected at 24 h after the last atomization, and the total number of karyocyte and the percentages of sorted cells were detected. The activity of interleukin-17 (IL-17) in BALF was detected by enzyme-linked immunosorbent assay (ELISA). Lung tissue was separated and subjected to hematoxylin-eosin (HE) staining, and the pathological changes of lung tissue were observed under an optical microscope. The proportion of T helper 17 (Th17) cells in the lung was detected by flow cytometry (FCM). After successful modeling of NA mice, immunomagnetic bead purified mouse splenic cluster of differentiation 4+(CD4+) T was treated with IL-7 and Ruxolitinib, and the proportion of differentiated Th17 cells to CD4+ T cells and Ki-67, B-cell lymphoma 2 (Bcl-2) and activated Caspase3 expressions in Th17 cells were detected via FCM.. Compared with those in NA group, the number of karyocytes and the percentages of neutrophils (NEU) and eosinophils (EOS) in BALF in Ruxo group were significantly reduced. The pathological changes of lung tissue in Ruxo group were overtly less than those in NA group. In comparison with NA group, Ruxo group had decreased IL-17A level in BALF and reduced proportion of Th17 cells in lung tissue. In in vitro experiment, compared with those in Con group, decreased percentages of Ki-67 and Bcl-2 proteins and increased percentage of Caspase3 in Th17 cells were found in Ruxo group.. Ruxolitinib may suppress the survival of Th17 cells by inhibiting the JAK/STAT5 signaling pathway and regulate the anti-apoptosis proteins Bcl-2 and Caspase3, thus promoting the increase of Thl7 cells entering the apoptotic pathway and reducing airway inflammation in NA mice.

Topics: Animals; Asthma; Disease Models, Animal; Female; Janus Kinases; Leukocyte Count; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Neutrophils; Nitriles; Ovalbumin; Pyrazoles; Pyrimidines; Random Allocation; Signal Transduction; STAT5 Transcription Factor

Asthma is characterized by airway hyperresponsiveness (AHR) and inflammation leading to airway remodeling (AR). In children, AR may occur very early prior to the age of 6 years. Treatments to prevent or reverse AR are unknown.. We sought to determine (i) whether short allergenic sensitization at a young age in a mouse model may induce enhanced AR and inflammation compared to adults; (ii) the effect of Montelukast on such AR.. Immature and adult Balb/c mice were sensitized and challenged with ovalbumin. AHR and AR were measured using cultured precision-cut lung slices and inflammation by bronchoalveolar lavage. Experiments were repeated after administration of Montelukast.. OVA-challenged mice developed AHR to methacholine regardless of age of first exposure to OVA. Young mice developed greater thickened basement membrane, increased smooth muscle mass, and increased area of bronchovascular fibrosis compared with adult mice. Cellular infiltrates in BAL differed depending upon animal age at first exposure with higher eosinophilia measured in younger animals. Montelukast decreased ASM mass, BAL cellularity.. We provide thus evidence for a greater degree of AR after allergenic sensitization and challenge in younger mice versus adults. This study provides proof of concept that airway remodeling can be prevented and reversed in this case by anti-asthmatic drug Montelukast in this model.

Topics: Acetates; Age Factors; Airway Remodeling; Allergens; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cyclopropanes; Disease Models, Animal; Female; Lung; Mice, Inbred BALB C; Ovalbumin; Quinolines; Sulfides

CD93 is a membrane-associated glycoprotein, which can be released in a soluble form (sCD93) into the serum. CD93 has received renewed attention as a candidate biomarker of inflammation in various inflammatory and immune-mediated diseases, including asthma.. We aimed to evaluate the effects of airway inflammation on CD93 levels in murine models.. We established an ovalbumin (OVA)-induced acute asthma murine model (OVA model) and a lipopolysaccharide (LPS)-induced airway inflammation murine model (LPS model). Dexamethasone was administered by gavage to attenuate the airway inflammation.. The OVA model demonstrated typical allergic asthma features with increased airway hyper-responsiveness, inflammatory cell infiltration, increased Th2 cytokine levels, compared to the control group. CD93 levels were decreased in lung homogenates and, respiratory epithelial cells, whereas serum sCD93 levels were increased in the OVA model, as compared to the control group. Dexamethasone reversed these effects of OVA. In contrast, in the LPS model, CD93 levels were not affected in neither respiratory epithelial cells nor serum.. Our findings demonstrate the potential of using sCD93 as a biomarker for allergic asthma.

Topics: Animals; Asthma; Biomarkers; Inflammation; Membrane Glycoproteins; Mice; Ovalbumin; Receptors, Complement

The purpose of this study is to explore the influence of ozone repeated short exposures on airway/lung inflammation, airway hyperresponsiveness (AHR) and airway hypersecretion in ovalbumin (OVA) sensitized/challenged asthmatic mouse model.. OVA sensitization was performing by intraperitoneal injection. Ozone exposures (3ppm for 3hours) were given one hour after aerosolized OVA challenges (once every other day, 4 times totally). Methacholine (MCH) bronchial provocation tests, Liu's staining of BALF cell smears, hematoxylin-eosin (HE) staining and Periodic Acid-Schiff (PAS) staining of lung tissue were performed. Interleukins (ILs; IL-4, IL-13, IL-1β, and IL-18) protein (ELISA) and mRNA expression levels (RT-qPCR) in murine lung, 8-hydroxy-2'-deoxyguanosine (8-OHdG, ELISA), malondialdehyde (MDA, thiobarbituric acid assay), reduced glutathione (GSH, spectrophotometric method) in bronchoalveolar lavage fluid (BALF), and GSH1 mRNA relative expression levels (RT-qPCR) in lung tissue were analyzed.. Repeated ozone exposures down-regulated the AHR to MCH in mice undergoing OVA sensitization and challenge, however not all parameters associated with asthma were decreased since obvious mucus hypersecretion was induced and airway inflammation increased slightly, especially around small airways. Following ozone co-exposure, the increase of IL-4 and IL-13 levels in murine lung caused by OVA sensitization/challenge were reversed. Instead, levels of IL-1β in BALF remained, higher than negative control group. Ozone repeated short exposures also induced significant increase of 8-OHdG in BALF in OVA sensitized and challenged mice.. For asthmatic mice undergoing ozone exposures, AHR is not an accurate indicator of the severity of asthma. Repeated short ozone exposures increase mucus hypersecretion, possibly via an increase in oxidative stress and immune dysregulation.

Topics: Animals; Asthma; Bronchial Hyperreactivity; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Interleukin-13; Interleukin-1beta; Interleukin-4; Male; Mice; Mice, Inbred C57BL; Mucus; Ovalbumin; Oxidative Stress; Ozone; Pneumonia; Reverse Transcriptase Polymerase Chain Reaction; Time Factors

Neutrophils are involved in the pathogenesis of allergy. However, the contribution of the different functionally polarized neutrophils in allergy needs to be clarified. We sought to define the characteristics of interleukin (IL)-33-induced neutrophils and the involvement of this subset of polarized neutrophils in allergic pathogenesis. Freshly isolated neutrophils were treated with different cytokines and the cytokine expression levels were detected by real-time PCR. The gene expression profile of IL-33-induced neutrophils was determined by microarray assay. Adoptive transfer assay was used to investigate the function of IL-33-induced neutrophils in an ovalbumin (OVA)-induced allergic asthma model. IL-33-treated neutrophils selectively produced IL-4, IL-5, IL-9 and IL-13 (referred as to N(IL-33) cells) and displayed a distinctive gene expression profile in sharp contrast to resting and lipopolysaccharide (LPS)-treated neutrophils. IL-33-induced neutrophils expressed high Levels of IL-1R2 on cell surface, whereas resting and LPS-treated neutrophils did not, indicating IL-1R2 might be used as a biomarker for N(IL-33) cells. Importantly, N(IL-33) neutrophils exist in the lungs of OVA-induced allergic asthma mice. Adoptive transfer of N(IL-33) neutrophils significantly promotes the severity of the lung pathogenesis in this model. IL-33 induces neutrophil polarization through c-Jun N-terminal kinase- and nuclear factor-κB-dependent pathways. A previously unappreciated neutrophil polarization driven by IL-33 with unique cell surface markers and cytokine/chemokine-producing gene profile was defined. The newly identified N(IL-33) subpopulation may have significant contribution to IL-33-related pathogenesis.

Topics: Alum Compounds; Animals; Asthma; Cell Polarity; Gene Expression Regulation; Inflammation; Interleukin-33; Lung; Male; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Models, Animal; Neutrophils; NF-kappa B; Ovalbumin; Receptors, Interleukin-1 Type II; Statistics, Nonparametric; Th2 Cells

Hyperactivation of phosphoinositol 3-kinase (PI3K) has been suggested to be a potential mechanism for endoplasmic reticulum (ER) stress-enhanced airway hyperresponsiveness, and PI3K inhibitors have been examined as asthma therapeutics. However, the regulatory mechanism linking PI3K to ER stress and related pathological signals in asthma have not been defined. To elucidate these pathogenic pathways, we investigated the influence of a selective PI3Kδ inhibitor, IC87114, on airway inflammation in an ovalbumin/lipopolysaccharide (OVA/LPS)-induced asthma model. In OVA/LPS-induced asthmatic mice, the activity of PI3K, downstream phosphorylation of AKT and activation of nuclear factor-κB (NF-κB) were all significantly elevated; these effects were reversed by IC87114. IC87114 treatment also reduced the OVA/LPS-induced ER stress response by enhancing the intra-ER oxidative folding status through suppression of protein disulfide isomerase activity, ER-associated reactive oxygen species (ROS) accumulation and NOX4 activity. Furthermore, inositol-requiring enzyme-1α (IRE1α)-dependent degradation (RIDD) of IRE1α was reduced by IC87114, resulting in a decreased release of proinflammatory cytokines from bronchial epithelial cells. These results suggest that PI3Kδ may induce severe airway inflammation and hyperresponsiveness by activating NF-κB signaling through ER-associated ROS and RIDD-RIG-I activation. The PI3Kδ inhibitor IC87114 is a potential therapeutic agent against neutrophil-dominant asthma.

Topics: Adenine; Animals; Asthma; Disease Models, Animal; Endoplasmic Reticulum Stress; Lipid Peroxidation; Lipopolysaccharides; Membrane Proteins; Mice; Nerve Tissue Proteins; NF-kappa B; Ovalbumin; Oxidation-Reduction; Oxidative Stress; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Quinazolines; Reactive Oxygen Species; Receptors, Cell Surface; Signal Transduction

IL-18 is likely to contribute to asthma. However, little is known regarding the role of IL-18 binding protein (BP) and IL-18 receptor (R) in asthma. Because the action of IL-18 in the body is regulated by IL-18BP and mast cells and basophils are key cell types involved in asthma, we investigated the expression of IL-18, IL-18BP and IL-18R in basophils and mast cells using flow cytometry and a mouse asthma model. We found that among basophils, approximately 53% and 51% were IL-18

Topics: Animals; Asthma; Basophils; Bronchoalveolar Lavage Fluid; Flow Cytometry; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-18; Interleukin-18 Receptor alpha Subunit; Lung; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Sputum

Allergic asthma is a complex and chronic inflammatory airway disease. The thymic stromal lymphopoietin (TSLP) signaling pathway plays an important role in asthma. Xiaoqinglong Decoction (XQL) is the first choice to treat cold asthma in clinical settings. In this study, the role of the TSLP pathway in the onset of asthma and the protective mechanism of XQL were investigated. A total of 50 female mice were randomly divided into the following groups: the blank group (A), the model group (B), the XQL group (C), the dexamethasone group (D), and the XQL + dexamethasone group (E). Asthma was induced with ovalbumin, and corresponding drug intervention was carried out for 7 days, after which serum and lung tissue end points were analyzed. Serum interleukin 1β (IL-1β), tumor necrosis factor-α (TNF-α), nuclear factor κB (NF-κB), and TSLP levels were higher in group B than in group A (p<0.05). However these levels were lower in group C and D than in group B (p<0.05), and there was no significant difference between groups C and D (p>0.05). Interestingly, these end points were significantly lower in group E than in groups C and D (p<0.05). Regarding pathologic changes, the inflammatory infiltrate in the lungs of groups C, D, and E was lower than that of group B, especially in group E. We conclude that the TSLP pathway plays an important role in the course of asthma, and can be used as an important target for asthma treatment; XQL may play a role in reducing inflammation and relieving asthma by regulating the TSLP signaling pathway.

Topics: Animals; Asthma; Complex Mixtures; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Female; Humans; Inflammation; Interleukin-1beta; Lung; Medicine, Chinese Traditional; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Signal Transduction; Thymic Stromal Lymphopoietin; Tumor Necrosis Factor-alpha

Connexion 43 (Cx43), a gap junction protein, is expressed abundantly in the airway and has been implicated in the pathogenesis of asthma. However, the effects of blocking Cx43 in asthma remain unclear. We investigated the therapeutic effects of two specific Cx43 inhibitors (Gap26 and Gap27) on the development of allergic airway disease in mice. Allergic asthma was induced in BALB/c mice by sensitization and challenge with ovalbumin (OVA). Different doses of Cx43 inhibitors were administered by aerosol inhalation 1 h after OVA challenge on days 21 and 23. Airway hyperresponsiveness (AHR), lung pathology, mucus production, and inflammatory cells and cytokines in bronchoalveolar lavage fluid (BALF) were examined. We found that Gap26 significantly inhibited OVA-induced AHR, inflammatory cell infiltration surrounding the bronchia, mucus production, inflammatory cells and cytokines in BALF, and OVA-specific IgE in the serum in a dose-dependent manner. Gap27 showed effects similar to those of Gap26 in inhibiting inflammatory cytokine production in BALF. We conclude Cx43 inhibitor inhalation alleviates asthma featuresin mice and may be a promising therapy for clinical asthma.

Topics: Animals; Asthma; Connexin 43; Cytokines; Disease Models, Animal; Female; Humans; Immunoglobulin E; Inflammation; Inflammation Mediators; Mice; Mice, Inbred BALB C; Ovalbumin; Peptides; Respiratory Hypersensitivity

Mucus production and hypersecretion are important pathophysiological features of asthma. Airway mucus secretion is more serious in obese asthma. Therefore, it is of great significance to elucidate the mechanism of asthma airway mucus high secretion in improving the control of asthma and the prognosis of obese asthmatic patients.. Obese asthmatic mice model was established to test the airway resistance and mucin secretion by hematoxylin-eosin (HE) staining. Munc18b and Muc5ac expression levels were determined by Western-blotting. Munc18b conditioned knockout mice were adopted to explore the mechanism of Muc5ac high secretion.. The mice weight increased in obese asthmatic model accompanied by elevated airway resistance. HE staining showed enhanced mucin secretion, which was correlated to weight and airway resistance. Munc18b and Muc5ac expressions significant upregulated in an obese asthmatic mouse model compared with normal control. Muc5ac expression failed to show elevation in Munc18b conditioned knockout mice.. Muc5ac high secretion was positively correlated with Munc18b upregulation in obese asthma. Munc18b participated in inducing Muc5ac high expression.

Topics: Animals; Asthma; Disease Models, Animal; Mice; Mice, Inbred C57BL; Mice, Knockout; Mucin 5AC; Munc18 Proteins; Obesity; Ovalbumin; Up-Regulation

Azithromycin is a potent agent that prevents airway remodeling. In this study, we hypothesized that azithromycin (35 mg/kg orally) alleviated airway remodeling through suppression of epithelial-to-mesenchymal transition (EMT) via downregulation of transforming growth factor-beta 1 (TGF-β1)/receptor for activated C-kinase1 (RACK1)/snail in mice. An ovalbumin (OVA)-induced Balb/c mice airway allergic inflammatory model was used. Airway inflammation and remodeling were evaluated with hematoxylin and eosin (HE), periodic acid-Schiff (PAS), and Masson staining. E-cadherin and N-cadherin (molecular markers of EMT) were analyzed by immunofluorescence, quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), and western blotting; α-smooth muscle actin (α-SMA) was evaluated using immunohistochemistry (IHC), qRT-PCR, and western blotting; and expression of TGF-β1/RACK1/Snail in lungs was measured by qRT-PCR and western blotting. Our data showed that azithromycin significantly reduced inflammation score, peribronchial smooth muscle layer thickness, goblet cell metaplasia, and deposition of collage fibers (P < 0.05), and effectively suppressed airway EMT (upregulated E-cadherin level, and downregulated N-cadherin and α-SMA levels) compared with the OVA group (P < 0.05). Moreover, the increasing mRNA and protein expressions of TGF-β1 and RACK1 and mRNA level of Snail in lung tissue were all significantly decreased in azithromycin-treated mice (P < 0.05). Taken together, our results suggest that azithromycin has the greatest effects on reducing airway remodeling by inhibiting TGF-β1/RACK1/Snail signal and improving the EMT in airway epithelium.

Topics: Airway Remodeling; Allergens; Animals; Anti-Bacterial Agents; Asthma; Azithromycin; Disease Models, Animal; Epithelial-Mesenchymal Transition; Humans; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors for Activated C Kinase; Respiratory Mucosa; Snail Family Transcription Factors; Transforming Growth Factor beta1

Asthma is a common chronic airway inflammation disease and is considered as a major public health problem. Fisetin (3,3',4',7-tetrahydroxyflavone) is a naturally occurring flavonoid abundantly found in different vegetables and fruits. Fisetin has been reported to exhibit various positive biological effects, including anti-proliferative, anticancer, anti-oxidative and neuroprotective effects. We evaluated the effects of fisetin on allergic asthma regulation in mice. Mice were first sensitized, then airway-challenged with ovalbumin (OVA). Whether fisetin treatment attenuated OVA-induced airway inflammation was examined via inflammation inhibition through MyD88-related NF-κB (p65) signaling pathway. Mice were divided into the control (Con), OVA-induced asthma (Mod), 40 (FL) and 50 (FH) mg/kg fisetin-treated OVA-induced asthma groups. Our results found that OVA-induced airway inflammation in mice caused a significant inflammatory response via the activation of MyD88 and NF-κB signaling pathways, leading to release of pro-inflammatory cytokines. In contrast, fisetin-treated mice after OVA induction inhibited activation of MyD88 and NF-κB signaling pathways, resulting in downregulation of pro-inflammatory cytokine secretion. Further, fisetin significantly ameliorated the airway hyperresponsiveness (AHR) towards methacholine (Mch). In addition, fisetin reduced the number of eosinophil, monocyte, neutrophil and total white blood cell in the bronchoalveolar lavage fluid (BALF) of OVA-induced mice. The serum and BALF samples obtained from the OVA-induced mice with fisetin showed lower levels of pro-inflammatory cytokines. The results of our study illustrated that fisetin may be a new promising candidate to inhibit airway inflammation response induced by OVA.

Topics: Airway Resistance; Animals; Asthma; Bronchial Hyperreactivity; Cell Count; Cytokines; Disease Models, Animal; Flavonoids; Flavonols; Inflammation Mediators; Lipopolysaccharides; Lung; Male; Mice, Inbred C57BL; Myeloid Differentiation Factor 88; NF-kappa B; Ovalbumin; Pneumonia; Signal Transduction; Toll-Like Receptor 5

Nicorandil is an antianginal drug that has anti-inflammatory property. This study aimed to investigate the effects of nicorandil on allergic asthma induced by ovalbumin (OVA) in mice in comparison with dexamethasone. Mice were sensitized to OVA (on days 0 and 7) and challenged with OVA three times (on days 14, 15 and 16). Nicorandil was given orally for 5 days 1 h before OVA treatment in days of challenge. Progression of asthma was accompanied by significant elevation in the lung/body weight index, LDH, total protein, IL-13 and NF-κB levels besides inflammatory cell counts in BALF; Also pulmonary MDA and NO contents were significantly increased but GSH and SOD levels were decreased. Histopathological alterations in lung tissues were also observed. In contrast, nicorandil treatment significantly alleviated OVA-induced lung injury. In conclusion, our results proposed that nicorandil is equivalent to dexamethasone in ameliorating allergic asthma by restoring oxidant/antioxidant balance and reducing inflammation.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Glutathione; Interleukin-13; Leukocyte Count; Lung; Male; Malondialdehyde; Mice; NF-kappa B; Nicorandil; Nitric Oxide; Ovalbumin; Superoxide Dismutase

Bronchial asthma is a chronic airway inflammatory disease which has three main pathological features: airway hyperresponsiveness (AHR), airway remodelling, and chronic inflammation. Acupuncture is known to be an effective integrative medical therapy that has been used in the treatment of several chronic diseases, including bronchial asthma. The aim of the current study was to evaluate the effects of acupuncture on inflammation and regulation of the IL-33/ST2 pathway in a mouse model of asthma.. The murine asthma model was established by both injection and inhalation of ovalbumin (OVA). Within 24 hours of the last OVA challenge, lung function was assessed by measurement of the airway resistance (R. The results showed that AHR, chronic inflammation and mucus secretion were significantly suppressed by acupuncture treatment. R. Acupuncture effectively protects lung function and attenuates airway inflammation in the OVA-induced mouse model of asthma, which supports the role of acupuncture as a potential therapy in asthma treatment.

Topics: Acupuncture Therapy; Animals; Asthma; Disease Models, Animal; Female; Humans; Interleukin-1 Receptor-Like 1 Protein; Interleukin-17; Interleukin-33; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Fcγ receptors (FcγR) play substantial immune regulatory roles both positively and negatively in pathophysiological processes including allergy and asthma. Compared with FcγRIIB which is classically defined as an inhibitory receptor, mouse FcγRIIIA and its functional human homologue FcγRIIA have been assumed to be activating receptors. However, evidence demonstrating inhibitory regulation by mouse FcγRIIIA has recently been emerging.. To dissect the contributory roles of mouse FcγRIIIA (human FcγRIIA) in parallel with FcγRIIB in an ovalbumin (OVA)-induced mouse model of asthma and to preliminarily assess the correlation of the respective FcγR with circulating IgE levels in human asthma patients.. Wild-type, FcγRIIB-/-, and FcγRIIIA-/- mice were used in an OVA-induced asthma model followed by assessment of the allergic pathology focused on the lung tissues. Expression levels of FcγRIIB, FcγRIIA, and FcγRIIIA on peripheral blood mononuclear cells (PBMC) together with the circulating IgE levels in the serum from patients with allergic asthma were analysed.. Although enhanced humoral immune responses typically represented by augmented OVA-specific IgG and IgE levels in serum were observed in the absence of FcγRIIIA in the mouse asthma model, no overall regulation by FcγRIIIA, especially in terms of those parameters measuring lung tissue inflammation, was recorded. As expected, in the absence of FcγRIIB, augmented immune responses measured as serum antibody levels as well as those in various regulatory pathways in this mouse asthma model were observed. The expression levels of human FcγRIIB but not FcγRIIA were negatively correlated with serum levels of IgE in human asthma patients.. We did not find major evidence demonstrating an immune inhibitory role of mouse FcγRIIIA in this OVA-induced mouse asthma model. As asthma is a complex disease and the immune regulatory responses involve sophisticated components and pathways, the exact roles of FcγRIIIA as well as its human functional homologue FcγRIIA in asthma still await further clarification using other mouse asthma models as well as clinical studies.

Topics: Allergens; Animals; Asthma; Child; Female; Flow Cytometry; Humans; Immunity, Humoral; Immunoglobulin E; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Pneumonia; Receptors, IgG; Reverse Transcriptase Polymerase Chain Reaction

Asthma is a chronic disease affecting up to 10% of the Australian population for which medical treatment is solely aimed at relief of symptoms rather than prevention of disease. Evidence from animal and human studies demonstrates a strong link between viral respiratory infections, atopy and the development of asthma. Type I IFNs include IFNα and IFNβ, with subtype expression tailored toward the specific viral infection. We hypothesized that exposure to type I IFNs and allergen may interfere with the healthy response to innocuous airway antigen exposure. In this study, we use an ovalbumin (OVA)-induced BALB/c model of experimental allergic airways disease, where pre-exposure of the airways to OVA is protective against allergen sensitization, leading to allergen tolerance. We investigated airways pre-exposure with OVA and type I IFNs on development of allergic airways disease. We demonstrate restoration of allergic airways disease on pre-exposure with allergen and IFNβ, and not IFNα. Dysfunction in tolerance led to changes in dendritic cell antigen capture/traffic, T-cell and B-cell responses. Furthermore, exposure to IFNβ with ongoing allergen exposure led to the development of hallmark asthma features, including OVA-specific IgE and airways eosinophilia. Data indicate a role for IFNβ in linking viral infection and allergy.

Topics: Allergens; Animals; Asthma; Disease Models, Animal; Eosinophils; Humans; Hypersensitivity; Immune Tolerance; Immunoglobulin E; Interferon-alpha; Interferon-beta; Lung; Mice; Mice, Inbred BALB C; Ovalbumin

Transient receptor potential canonical channel 3 (TRPC3) proteins function as non-voltage-gated Ca

Topics: Acetylcholine; Airway Remodeling; Aminophylline; Animals; Antibodies; Asthma; Calcium; Cell Proliferation; Cells, Cultured; Dexamethasone; Gadolinium; Hydroxyproline; Mice; Muscle Contraction; Muscle, Smooth; Ovalbumin; Protein Biosynthesis; RNA Interference; RNA, Small Interfering; TRPC Cation Channels

Cold air stimulus is an important environmental factor that exacerbates asthma. At the molecular level, the transient receptor potential melastatin 8 (TRPM8) plays a crucial part in cold detection. The roles of TRPM8 in airway inflammation and remodeling in a murine model of asthma with cold stimulus and the related molecular mechanism are largely unknown. In this study, C57BL/6 mice were randomly divided into four groups: phosphate-buffered saline control group (control), ovalbumin (OVA)-induced asthma group (OVA), OVA with cold air stimulus group (OVA+cold), and OVA+cold+shTRPM8 (TRPM8 short hairpin RNA) group. We showed that cold air stimulus-induced TRPM8 upregulation in the OVA+cold group. Moreover, TRPM8 knockdown significantly attenuated cold-induced inflammation and infiltration, decreased levels of immunoglobulin E, restored the Th1/Th2 balance, and reduced inflammatory cell accumulation and airway remodeling. Furthermore, we demonstrated that TRPM8 knockdown dramatically inhibited mitogen-activated protein kinase and nuclear factor-κB pathways. Collectively, these results revealed that cold air stimulus induced an airway inflammatory response and remodeling by increasing TRPM8 expression and that downregulation of TRPM8 alleviated these responses.

Topics: Air; Airway Remodeling; Animals; Asthma; Cold Temperature; Cytokines; Immunoglobulin E; Mice, Inbred C57BL; Mitogen-Activated Protein Kinases; NF-kappa B; Ovalbumin; Pneumonia; Random Allocation; RNA Interference; Signal Transduction; TRPM Cation Channels

Cassia occidentalis Linn. is a traditional ayruvedic edible shrub containing anthraquinones (AQs) as the principle active constituents. In folk medicine, it has a variety of uses including treatment of whooping cough ('pertussis') and inflammatory diseases. Despite these applications, limited data are available to validate the effects of C. occidentalis AQs on airways inflammation in asthma.. To explore the anti-inflammatory potential of AQs extracted from C. occidentalis using an in vivo model of ovalbumin (OVA)-induced asthma.. Extraction and optimization of AQs from C. occidentalis was performed by mechanochemistry. Allergic asthma in BALB/c mice was sensitized and challenged by OVA, and the effects of AQs investigated in a mouse model. OVA-specific IgE concentrations in serum, and Th1/Th2 cytokine (IL-4, IL-5, IL-13 and IFN-γ) concentrations, inflammatory cell counts and classification in bronchoalveolar lavage fluid (BALF) were determined. Histopathological evaluation of lung tissue was performed using hematoxylin and eosin (H&E), and periodic acid-schiff (PAS) staining. Th1/Th2 cytokine mRNA expression was analyzed using the 2. Treatment with AQs decreased inflammatory cell counts and production of Th2 cytokines (IL-4, IL-5 and IL-13) in BALF, and OVA-specific IgE in serum. In contrast,Th1 cytokine IFN-γ production in BALF was promoted. AQs also decreased mRNA expression of Th1/Th2 cytokine in lung tissue. Histological studies demonstrated that AQs substantially inhibited OVA-induced cellular infiltration, mucus hypersecretion and goblet cell hyperplasia in the lung.. These findings demonstrated the inhibitory effects of AQs, derived from C. occidentalis, on OVA-induced allergic asthma in mice. The results suggest a promising ethnopharmacological use for AQs in patients with asthma.

Topics: Allergens; Animals; Anthraquinones; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cell Survival; Cytokines; Disease Models, Animal; Female; Immunoglobulin E; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Plant Components, Aerial; RAW 264.7 Cells; RNA, Messenger; Senna Plant

The experimental model of combined allergic rhinitis and asthma syndrome (CARAS) has shown that CpG oligodeoxynucleotides (CpG-ODNs) are potential inhibitors of type 2 helper cell-driven inflammatory responses. Currently available CpG-ODNs modestly inhibit allergic responses in CARAS, while a combination strategy for upper airway treatment by co-administration of CpG-ODNs and glucocorticoids may show good efficacy. This study aimed to assess the therapeutic effects of CpG-ODNs combined with budesonide (BUD) on upper and lower-airway inflammation and remodeling in mice with CARAS induced by chronic exposure to ovalbumin (OVA), exploring the possible underlying molecular mechanisms. A BALB/c mouse model of chronic CARAS was established by systemic sensitization and repeated challenge with OVA. Treatment with CpG-ODNs or BUD by intranasal administration was started 1 h after OVA challenge. Then, nasal mucosa and lung tissues were fixed and stained for pathologic analysis. The resulting immunologic variables and TSLP-DC-OX40L axis parameters were evaluated. Both CpG-ODNs and BUD intranasal administration are effective on reducing Th2-type airway inflammation and tissue remodeling. Co-administration of CpG-ODNs and BUD was more effective than each monotherapy in attenuating upper and lower-airway inflammation as well as airway remodeling in chronic CARAS. Notably, combination of CpG-ODNs with BUD modulated the TSLP-DC-OX40L axis, as demonstrated by decreased TSLP production in the nose and lung, alongside decreased TSLPR and OX40L in DC. Intranasal co-administration of CpG-ODNs and BUD synergistically alleviates airway inflammation and tissue remodeling in experimental chronic CARAS, through shared cellular pathways, as a potent antagonist of the TSLP-DC-OX40L axis.

Topics: Airway Remodeling; Animals; Asthma; Budesonide; Cytokines; Drug Synergism; Inflammation; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Oligodeoxyribonucleotides; Ovalbumin; OX40 Ligand; Rhinitis, Allergic; Thymic Stromal Lymphopoietin; Tumor Necrosis Factor Inhibitors

Siglec-F is a pro-apoptotic receptor on mouse eosinophils that recognizes 6'-sulfated sialyl Lewis X and 6'-sulfated sialyl N-acetyl-lactosamine as well as multivalent sialyl N-acetyl-lactosamine structures on glycan arrays. We hypothesized that attenuation of the carbohydrate sulfotransferase 1 (CHST1) gene encoding keratan sulfate galactose 6-O-sulfotransferase, an enzyme likely required for 6'-sulfation of some of these putative Siglec-F glycan ligands, would result in decreased Siglec-F lung ligand levels and enhanced allergic eosinophilic airway inflammation. Tissue analysis detected CHST1 expression predominantly not only in parenchymal cells but not in airway epithelium, the latter being a location where Siglec-F ligands are located. Western blotting of lung extracts with Siglec-F-Fc fusion proteins detected ≈500 kDa and ≈200 kDa candidate Siglec-F ligands that were not appreciably altered in CHST1-/- lungs compared with normal mouse lungs. Characterization of the O-linked glycans of lung tissue and bronchoalveolar lavage fluid detected altered sialylation but minimal change in sulfation. Eosinophilic airway inflammation was induced in wild-type (WT) and CHST1-/- mice via sensitization to ovalbumin (OVA) and repeated airway challenge. After OVA sensitization and challenge, Siglec-F ligands on airway cells, and numbers of eosinophils and neutrophils accumulating in the airways, both increased to a similar degree in WT and CHST1-/- mouse lungs, while macrophages and lymphocytes increased significantly more in CHST1-/- mouse airway compared with normal mouse lungs. Therefore, keratan sulfate galactose 6-O-sulfotransferase does not contribute to the synthesis of glycan ligands for Siglec-F in the airways, although its absence results in exaggerated accumulation of airway macrophages and lymphocytes.

Topics: Animals; Antigens, Differentiation, Myelomonocytic; Asthma; Carbohydrate Sulfotransferases; Lung; Lymphocytes; Macrophages; Mice; Mice, Inbred C57BL; Ovalbumin; Polysaccharides; Respiratory Mucosa; Sialic Acid Binding Immunoglobulin-like Lectins; Sulfotransferases

Thirdhand smoke (THS) represents the accumulation of secondhand smoke on indoor surfaces and in dust, which, over time, can become more toxic than secondhand smoke. Although it is well known that children of smokers are at increased risk for asthma or asthma exacerbation if the disease is already present, how exposure to THS can influence the development or exacerbation of asthma remains unknown.. We investigated whether epicutaneous exposure to an important component of THS, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), can influence asthma pathology in a mouse model elicited by means of repeated intranasal challenge with cockroach antigen (CRA).. Wild-type mice, α7 nicotinic acetylcholine receptor (nAChR)- or mast cell (MC)-deficient mice, and mice with MCs that lacked α7 nAChRs or were the host's sole source of α7 nAChRs were subjected to epicutaneous NNK exposure, intranasal CRA challenge, or both, and the severity of features of asthma pathology, including airway hyperreactivity, airway inflammation, and airway remodeling, was assessed.. We found that α7 nAChRs were required to observe adverse effects of epicutaneous NNK exposure on multiple features of CRA-induced asthma pathology. Moreover, MC expression of α7 nAChRs contributed significantly to the ability of epicutaneous NNK exposure to exacerbate airway hyperreactivity to methacholine, airway inflammation, and airway remodeling in this model.. Our results show that skin exposure to NNK, a component of THS, can exacerbate multiple features of a CRA-induced model of asthma in mice and define MCs as key contributors to these adverse effects of NNK.

Topics: Administration, Intranasal; Allergens; alpha7 Nicotinic Acetylcholine Receptor; Animals; Asthma; Cockroaches; Cytokines; Disease Models, Animal; Female; Lung; Mast Cells; Mice, Inbred C57BL; Mice, Transgenic; Nitrosamines; Ovalbumin; Skin; Smoke; Tobacco Smoke Pollution

Previous studies have shown that lipopolysaccharide (LPS) exposure may have a protective effect on asthma by reducing airway hyper-responsiveness, airway inflammation and serum IgE levels. However, there are few studies investigating the effect of LPS on mucous secretion in asthma. In this study, we evaluate the relationship between LPS pre-treatment in infant mice and airway mucus hypersecretion in an OVA (ovalbumin)-induced asthma model, and further explore the mechanisms behind this effect.. Mice were pre-treated with LPS by intranasal instillation (i.n.) from the 3rd day of life for 10 consecutive days before the OVA-induced asthma model was established. In order to detect mucus secretion, periodic acid-Schiff (PAS) staining was carried out, and the expression of Muc5ac was detected. The IL-13 levels in Bronchoalveolar lavage fluid (BALF) and lung tissue were also detected. In vitro, the expression of Muc5ac mRNA and protein was quantified in IL-13-stimulated 16HBE cells with or without LPS pre-treatment. In addition, proteins in the JAK2/STAT6 pathway, transcription factors (forkhead box transcription factor A2 (FOXA2), activation protein-1(AP-1), NF-κB), and the levels of reactive oxygen species (ROS) were also measured in vivo and in vitro.. LPS pre-treatment reduced mucus secretion, as demonstrated by decreased PAS staining and muc5ac expression. Further exploration of the underlying mechanisms of this phenomenon revealed that LPS pre-treatment decreased the production of IL-13, IL-13 induced ROS synthesis was reduced, and the JAK2/STAT6 pathway was inhibited. Decreased stat6 increased transcription factor FOXA2, and the relatively increased FOXA2 further decreased the level of Muc5ac and mucous hypersecretion in OVA-induced asthma.. LPS pre-treatment ameliorated mucus hypersecretion in an OVA-induced asthma model by inhibition of IL-13 production and by further inhibiting the JAK2/STAT6 pathway and ROS activity, and up-regulating expression of FOXA2.

Topics: Administration, Intranasal; Animals; Asthma; Cell Line; Disease Models, Animal; Down-Regulation; Humans; Interleukin-13; Janus Kinases; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Mucin 5AC; Mucus; Ovalbumin; Protective Agents; Signal Transduction; STAT6 Transcription Factor

Patients with DOCK8 deficiency are at increased susceptibility to develop allergic diseases such as food allergy and asthma. Here, we aimed to analyze the pathogenesis of asthma in DOCK8-deficient patients. In our mouse model, DOCK8-knockout (KO) mice sensitized with low-dose OVA were challenged with 1.5% OVA to induce allergic asthma. As compared to that in WT mice, remarkable airway hyperresponsiveness was observed in KO mice. Increased inflammatory cells and eosinophils infiltrated in airway lumen in KO mice especially around bronchi. KO mice showed higher levels of serum IgE and OVA-specific IgE and significantly elevated IgE-producing B cells in blood and in spleen. Surprisingly, nasal administration with rmIL-21 significantly reduced the airway hyperresponsiveness, inflammatory infiltration, as well as the serum IgE and IgE-producing B cells. DOCK8-knockout mice are susceptible to low-dose OVA induced allergic airway inflammation and airway hyperresponsiveness. Supplementary nasal administration of rmIL-21 alleviates allergic asthma in this mouse model.

Topics: Administration, Intranasal; Animals; Asthma; B-Lymphocytes; Disease Models, Animal; Guanine Nucleotide Exchange Factors; Immunoglobulin E; Interleukins; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Recombinant Proteins; Respiratory Hypersensitivity

Ras, a small GTPase protein, is thought to mediate Th2-dependent eosinophilic inflammation in asthma. Ras requires cell membrane association for its biological activity, and this requires the posttranslational modification of Ras with an isoprenyl group by farnesyltransferase (FTase) or geranylgeranyltransferase (GGTase). We hypothesized that inhibition of FTase using FTase inhibitor (FTI)-277 would attenuate allergic asthma by depleting membrane-associated Ras. We used the OVA mouse model of allergic inflammation and human airway epithelial (HBE1) cells to determine the role of FTase in inflammatory cell recruitment. BALB/c mice were first sensitized then exposed to 1% OVA aerosol or filtered air, and half were injected daily with FTI-277 (20 mg/kg per day). Treatment of mice with FTI-277 had no significant effect on lung membrane-anchored Ras, Ras protein levels, or Ras GTPase activity. In OVA-exposed mice, FTI-277 treatment increased eosinophilic inflammation, goblet cell hyperplasia, and airway hyperreactivity. Human bronchial epithelial (HBE1) cells were pretreated with 5, 10, or 20 μM FTI-277 prior to and during 12 h IL-13 (20 ng/ml) stimulation. In HBE1 cells, FTase inhibition with FTI-277 had no significant effect on IL-13-induced STAT6 phosphorylation, eotaxin-3 peptide secretion, or Ras translocation. However, addition of exogenous FPP unexpectedly augmented IL-13-induced STAT6 phosphorylation and eotaxin-3 secretion from HBE1 cells without affecting Ras translocation. Pharmacological inhibition of FTase exacerbates allergic asthma, suggesting a protective role for FTase or possibly Ras farnesylation. FPP synergistically augments epithelial eotaxin-3 secretion, indicating a novel Ras-independent farnesylation mechanism or direct FPP effect that promotes epithelial eotaxin-3 production in allergic asthma.

Topics: Animals; Asthma; Bronchi; Bronchial Hyperreactivity; Disease Models, Animal; Enzyme Inhibitors; Eosinophils; Epithelial Cells; Farnesyltranstransferase; Humans; Inflammation; Lung; Male; Methionine; Mice; Mice, Inbred BALB C; Ovalbumin; Polyisoprenyl Phosphates; ras Proteins; Sesquiterpenes; Signal Transduction

Persistent inflammation and remodeling of airways are the major hallmarks of asthma. Though airway inflammation diminishes in ovalbumin (OVA)-based mouse model of chronic asthma owing to immune-tolerance linked with repeated allergen exposure, which limits the application of the disease model. Accordingly, the present study was designed to develop a murine model of chronic asthma which presents persistent airway inflammation coupled with remodeling traits. Herein, OVA-sensitized BALB/c mice were challenged with increasing (modified protocol) or constant concentration (conventional protocol) of the allergen for 6 weeks; 3 times/week. The results, indeed, revealed that mice subjected to modified protocol demonstrate an improved response to the allergen as reflected by the significant increase in inflammatory cells particularly, eosinophils in bronchoalveolar lavage fluid compared to conventional protocol. Moreover, the expression of Th2 cytokines and their responsible transcription factors (GATA-3 and STAT-6) was markedly enhanced in lungs. The increase in inflammation was further accompanied by a marked increase in mucus production, collagen deposition, and the expression of allied factors (Muc5ac, Col1α1, and α-SMA). Interestingly, pre-treatment of dexamethasone, a corticosteroid (0.5 mg/kg b.wt., i.p.), suppressed the allergen-induced airway inflammation and mucus production without altering collagen deposition. Failure of dexamethasone seems to be related to their ineffectiveness to modulate the expression of TGF-β, MMP-9, COL1α1, and α-SMA. Overall, our results strongly suggest that mice underwent modified chronic protocol bears more resemblance with asthmatics as it imitates persistent airway inflammation allied with steroid-refractory remodeling traits; hence, may be useful for the evaluation of new/alternative drugs in steroid-refractory asthmatic conditions.

Topics: Acute Disease; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Chronic Disease; Cytokines; Disease Models, Animal; Immune Tolerance; Leukocyte Count; Lung; Male; Mice, Inbred BALB C; Ovalbumin; RNA, Messenger

We previously reported that the biologically active form of histamine releasing factor (HRF) is dimerized translationally controlled tumor protein (dTCTP) which is involved in a number of allergic diseases.. Hoping that agents that modulate dTCTP may provide new therapeutic targets to allergic inflammatory diseases, we screened a library of natural products for substances that inhibit dTCTP. One such inhibitor we found was dehydrocostus lactone (DCL), a natural sesquiterpene present in rhizome of Saussurea lappa Clarke, the subject of this study.. We evaluated the therapeutic efficacy of DCL in a mouse model of ovalbumin (OVA)-induced allergic airway inflammation, employing the ELISA system using BEAS-2B cells and splenocytes, and confirmed that DCL interacts with dTCTP using SPR assay.. DCL inhibited dTCTP-induced secretion of IL-8 in BEAS-2B cells. From kinetic analysis of dTCTP and DCL, we found that K. DCL's therapeutic potential in allergic airway inflammation is based on its anti-inflammatory activity of suppressing the function of dTCTP.

Topics: Animals; Asthma; Biomarkers, Tumor; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Hypersensitivity; Immunoglobulin E; Inflammation; Kinetics; Lactones; Mice, Inbred BALB C; Ovalbumin; Protein Multimerization; Saussurea; Sesquiterpenes; Surface Plasmon Resonance; Tumor Protein, Translationally-Controlled 1

BACKGROUND It is generally believed that endotoxin exposure exacerbates risk of developing asthmatic symptoms. However, recent studies have indicated that prior bacterial exposure may prevent future symptoms of asthma. Here, we evaluated the influence of pre-exposure to different concentrations of lipopolysaccharide (LPS) to subsequent ovalbumin (OVA) allergen sensitization and challenge. MATERIAL AND METHODS Four-week-old Balb/c mice were treated intranasally with varying concentrations of LPS (1 ug, 10 ug, and 100 ug) or sterile PBS for 10 days, then 2 weeks later they were exposed to OVA. Both the molecular and functional airway responses to OVA administration were assessed following prior exposure to different doses of LPS or controls. Additionally, the Th1/Th2 and Treg/Th17 balance was measured. RESULTS Airway responsiveness and immune cell recruitment in the bronchoalveolar lavage (BALF) were decreased in animals exposed to a low dose of LPS (1 ug) treatment compared with the asthma group. Moderate-dose (10 ug) and high-dose (100 ug) LPS administration showed no differences from controls. Further, low-dose LPS (1 ug) exposure was associated with increased Th1 cytokines, T-bet, Treg cytokine (IL-10, TGF-β), and Foxp3 expression, but decreased Th2 cytokines (IL-4,5,13), GATA3, Th17, and ROR-gt expression compared with the asthma group. Finally, higher numbers of CD4+CD25+Foxp3+Treg cells, and CD4+INF-γ+T cells, and lower CD4+IL-4+T cells and CD4+IL-17+T cells were observed in the low-dose LPS-treated groups compared to controls. CONCLUSIONS Our findings suggest that prior exposure to low doses of LPS may protect from OVA-induced airway hyperresponsiveness (AHR) and histopathologic changes through regulation of the Th1/Th2 and Treg/Th17 balance.

Topics: Animals; Antigens, CD; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Gene Expression Regulation; Granulocytes; Lipopolysaccharides; Lung; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; RNA, Messenger; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Th2 Cells

We investigated whether the therapeutic effects of dexamethasone for allergic asthma and rhinitis were enhanced in mice when exposed to hypergravity. Forty mice were divided into 5 groups (n = 8/group): Control group received saline intraperitoneally (i.p.) and intranasally (i.n.); Asthma group received i.p./i.n. ovalbumin (OVA) for inducing allergic asthma/rhinitis; Dexa group received i.n. dexamethasone (0.75 mg/kg) 30 minutes before each OVA challenge; Hypergravity group was subjected to allergic asthma/rhinitis as well as exposed to 5 G hypergravity for 30 days; Finally in Dexa/Hypergravity group, hypergravity and dexamethasone were used simultaneously during induction of allergic asthma/rhinitis. Dexa group and Hypergravity group showed a significant decrease in serum total IgE levels compared to the Asthma group (p<0.05). Dexa/Hypergravity group showed greater IgE decrease compared with Dexa group (p = 0.040). Compared with the monotherapy groups, Dexa/Hypergravity group showed significantly fewer eosinophils in BAL fluid (p<0.05). Dexa/Hypergravity group showed significantly decreased eosinophilic infiltration into the lungs and nasal cavity (p<0.05). EC-SOD (extracellular superoxide dismutase) expression was significantly upregulated in the Hypergravity group and Dexa/Hypergravity group, compared with the Dexa group (p<0.05). In conclusion, hypergravity enhanced the therapeutic effect of dexamethasone in a murine model of allergic asthma and rhinitis. Therefore, combination could be a promising strategy, and one of its mechanisms could be up-regulation of EC-SOD expression.

Topics: Animals; Anti-Allergic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Dexamethasone; Drug Evaluation, Preclinical; Enzyme Induction; Eosinophils; Female; Hypergravity; Lung; Lymphocytes; Mice; Mice, Inbred BALB C; Nasal Cavity; Neutrophils; Ovalbumin; Rhinitis, Allergic; Specific Pathogen-Free Organisms; Superoxide Dismutase; Up-Regulation

Exosomes are nanovesicles originating from multivesicular bodies that have complex functions and significant therapeutic effects in many diseases. In the present study, we successfully extracted exosomes from Pseudomonas aeruginosa and assessed the effect of those exosomes on the development of the allergic response in two types of classic asthma models.. Female BALB/c mice were administrated with P. aeruginosa-derived exosomes 1 week before ovalbumin (OVA) or house dust mite (HDM) sensitization. Bronchoalveolar lavage fluid, serums and lung tissues were collected and analyzed for pathophysiology and immune responses.. Our results demonstrated that P. aeruginosa-derived exosomes inhibited the development of airway hyper-responsiveness (AHR), peribronchial and perivascular inflammation in lung tissues and the level of serum IgE. Moreover, this protective effect was associated with an increase in the regulatory T cell (T. In conclusion, these observations demonstrated that P. aeruginosa-derived exosomes could induce protection against allergic sensitization in asthma mice, and our study provided a new insight to prevent allergic diseases.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Exosomes; Female; Hypersensitivity; Immune System; Immunoglobulin E; Inflammation; Lipopolysaccharides; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Pseudomonas aeruginosa; T-Lymphocytes, Regulatory

While calcitriol can inhibit airway remodeling in asthmatic mice, the mechanism remains unclear. The purpose of this study was to explore the mechanism of action of calcitriol on airway remodeling in asthma and its interaction with budesonide.. A mouse model of asthma was established by allergic sensitization and challenge with ovalbumin. The mice were treated with budesonide, calcitriol, or budesonide plus calcitriol. The expression of airway remodeling-related proteins, transforming growth factor. Monotherapy with budesonide or calcitriol inhibited the high expression of collagen type I protein and upregulated the low expression of Smad7 in asthmatic mice. There was a synergistic interaction between budesonide and calcitriol in combined treatment. The expression of miR-21 in the combined treatment group was significantly lower than that in the calcitriol treatment group. VDR expression in the combined treatment group was significantly higher than that of the calcitriol treatment group.. Budesonide and calcitriol have a synergistic effect on airway remodeling in asthmatic mice.

Topics: Airway Remodeling; Animals; Anti-Inflammatory Agents; Asthma; Budesonide; Calcitriol; Calcium-Regulating Hormones and Agents; Disease Models, Animal; Drug Evaluation, Preclinical; Drug Synergism; Female; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Random Allocation; Receptors, Calcitriol; Receptors, Glucocorticoid; Retinoid X Receptors; Smad Proteins; Transforming Growth Factor beta

Membrane-associated RING-CH-1 (March1) is a member of the March family of E3 ubiquitin ligases. March1 downregulates cell surface expression of MHC II and CD86 by targeting them to lysosomal degradation. Given the key roles of MHC class II and CD86 in T cell activation and to get further insights into the development of allergic inflammation, we asked whether March1 deficiency exacerbates or attenuates features of allergic asthma in mice. Herein, we used an acute model of allergy to compare the asthmatic phenotype of March1-deficient and -sufficient mice immunized with ovalbumin (OVA) and later challenged by intranasal instillation of OVA in the lungs. We found that eosinophilic inflammation in airways and lung tissue was similar between WT and March1

Topics: Allergens; Animals; Asthma; Cytokines; Disease Models, Animal; Humans; Immunoglobulin E; Lung; Lymphocyte Activation; Mice; Mice, Inbred C57BL; Mice, Knockout; Neutrophil Infiltration; Ovalbumin; Pneumonia; Ubiquitin-Protein Ligases

To investigate the role of toll-like receptor 2 (TLR2) in asthmatic mouse model and its possible signal transduction pathways.. Mice were divided into three groups: TLR2-/- asthma mouse model group (n=10), C57BL/6 asthma mouse model group (n=10) and control group (n=10). Mice were sensitized and stimulated with ovalbumin (OVA) to establish the asthmatic mouse model. The unilateral bronchoalveolar lavage fluid (BALF) was collected and centrifuged to separate cells, and the cells were classified and counted via smear test under a microscope. Part of the lung tissues on the other side was taken for hematoxylin-eosin (HE) staining to observe the histopathological change in lung tissues. The remaining lung tissues on the other side were taken to detect the messenger ribonucleic acid (mRNA) expression levels of interleukin-4 (IL-4), IL-5 and IL-13 via reverse transcription-polymerase chain reaction (RT-PCR). The levels of nuclear factor-κB (NF-κB) p65, phosphorylated (p)-NF-κB p65, p-IκBα, extracellular signal-regulated kinase (ERK)1/2, p-ERK1/2, Jun N-terminal kinase (JNK), p-JNK, p38 mitogen-activated protein kinase (MAPK), p-p38 MAPK, IL-4, IL-5 and IL-13 were detected via enzyme-linked immunosorbent assay (ELISA). The protein expressions of NF-κB p65, p-NF-κB p65, p-IκBα, ERK1/2, p-ERK1/2, JNK, p-JNK, p38 MAPK, and p-p38 MAPK were detected using the immunohistochemical method.. HE staining showed that the infiltration degree of inflammatory cells in perivascular tissues in TLR2-/- asthma group was reduced compared with that in C57BL/6 asthma group. Results of electron microscopy showed that the ultrastructural changes in alveolar type I epithelial cells in mice in TLR2-/- asthma group was significantly alleviated. In BALF in TLR2-/- asthma group, the numbers of eosinophils and lymphocytes were significantly decreased, but the number of macrophages was significantly increased compared with those in C57BL/6 asthma group. Results of RT-PCR and ELISA revealed that the mRNA and protein expression levels of IL-4, IL-5, and IL-13 in lung tissues of mice in TLR2-/- asthma group were significantly decreased compared with those in C57BL/6 asthma group. Besides, results of ELISA and immunohistochemistry revealed that the protein expressions of NF-κB p65, p-NF-κB p65, p-IκBα, ERK1/2, JNK, p38 MAPK, p-ERK1/2, p-JNK, and p-p38 MAPK in lung tissues of mice in TLR2-/- asthma group were significantly decreased compared with those in C57BL/6 asthma group.. TLR2 is involved in the occurrence and development of experimental asthmatic airway inflammation. TLR2 gene knockout in asthmatic mice can alleviate the airway inflammation, whose mechanism may be that the allergic airway inflammation of asthmatic mice is alleviated through inhibiting NF-κB and MAPK signaling pathways.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Gene Knockdown Techniques; Inflammation; Lung; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; NF-kappa B; Ovalbumin; Toll-Like Receptor 2

Bronchial asthma is characterized by type 2 T helper (Th2) cell inflammation, essentially due to a breakdown of immune tolerance to harmless environmental allergens. Etiologically, experiences of psychological stress can be associated with a heightened prevalence of asthma. However, the mechanisms underlying stress-related asthma development are unclear. In this study, we examined whether psychological stress increases susceptibility to allergic asthma by downregulating immune tolerance.. Female BALB/c mice were sensitized with ovalbumin/alum, followed by ovalbumin inhalation. Ovalbumin inhalation induced immune tolerance before sensitization occurred. Some mice were exposed to restraint stress during tolerance induction or sensitization. Asthma development was evaluated by airway responsiveness, inflammation, cytokine expression, and IgE synthesis. Sensitization was evaluated by measuring proliferation and cytokine production by splenocytes. The effects of stress exposure on the numbers and functions of dendritic cells and regulatory T (Treg) cells in bronchial lymph nodes and spleens were evaluated. To investigate the role of endogenous glucocorticoid in inhibiting immune tolerance after stress exposure, we examined the effects of (i) a glucocorticoid-receptor antagonist administered prior to stress exposure, and (ii) exogenous gluco-corticoid (instead of stress exposure).. Asthmatic responses and Th2-biased sensitization, which were suppressed in tolerized mice, re-emerged in tolerized mice stressed during tolerance induction in association with decreased tolerogenic dendritic and Treg cell numbers. The effects of stress exposure on tolerized mice were abolished by administering a glucocorticoid-receptor antagonist and reproduced by administering exogenous glucocorticoid without stress.. Our findings suggested that psychological stress can potentially increase allergic asthma susceptibility by inhibiting immune tolerance.

Topics: Adoptive Transfer; Allergens; Alum Compounds; Animals; Asthma; Biomarkers; Corticosterone; Cytokines; Dendritic Cells; Disease Susceptibility; Female; Immune Tolerance; Immunization; Immunoglobulin E; Mice; Mice, Knockout; Ovalbumin; Receptors, Glucocorticoid; Respiratory System; Spleen; Stress, Psychological; T-Lymphocyte Subsets; Th2 Cells

We have previously shown that domperidone-induced short-term hyperprolactinemia reduces the lung's allergic inflammatory response in an ovalbumin antigenic challenge model. Since purinergic receptor P2X7R activity leads to proinflammatory cytokine release and is possibly related to the pathogenesis of allergic respiratory conditions, the present study was designed to investigate a possible involvement of purinergic and prolactin receptors in this phenomenon.. To induce hyperprolactinemia, domperidone was injected intraperitoneally in rats at a dose of 5.1 mg × kg-1 per day for 5 days. P2X7 expression was evaluated by lung immunohistochemistry while prolactin receptor expression in bronchoalveolar lavage leukocytes was analyzed through flow cytometry.. Previous reports demonstrated that rats subjected to short-term hyperprolactinemia exhibited a decrease in leukocyte counts in bronchoalveolar lavage, especially granulocytes. Here, it is revealed that hyperprolactinemia promotes an increased expression of prolactin receptors in granulocytes. Also, increased expression of purinergic P2X7R observed in allergic animals was significantly reduced by hyperprolactinemia.. Both purinergic and prolactin receptor expression changes occur during the anti-asthmatic effect of hyperprolactinemia.

Topics: Animals; Asthma; Gene Expression; Hyperprolactinemia; Leukocyte Count; Lung; Male; Ovalbumin; Rats; Rats, Wistar; Receptors, Purinergic P2X7; Time Factors

Allergic asthma is a chronic inflammatory disease dominated by a CD4+ T helper 2 (Th2) cell signature. The immune response amplifies in self-enforcing loops, promoting Th2-driven cellular immunity and leaving the host unable to terminate inflammation. Posttranscriptional mechanisms, including microRNAs (miRs), are pivotal in maintaining immune homeostasis. Since an altered expression of various miRs has been associated with T cell-driven diseases, including asthma, we hypothesized that miRs control mechanisms ensuring Th2 stability and maintenance in the lung. We isolated murine CD4+ Th2 cells from allergic inflamed lungs and profiled gene and miR expression. Instead of focusing on the magnitude of miR differential expression, here we addressed the secondary consequences for the set of molecular interactions in the cell, the interactome. We developed the Impact of Differential Expression Across Layers, a network-based algorithm to prioritize disease-relevant miRs based on the central role of their targets in the molecular interactome. This method identified 5 Th2-related miRs (mir27b, mir206, mir106b, mir203, and mir23b) whose antagonization led to a sharp reduction of the Th2 phenotype. Overall, a systems biology tool was developed and validated, highlighting the role of miRs in Th2-driven immune response. This result offers potentially novel approaches for therapeutic interventions.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Gene Expression Profiling; Gene Expression Regulation; Gene Regulatory Networks; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Primary Cell Culture; Protein Interaction Maps; Systems Biology; Th2 Cells

Dendritic cell (DC) trafficking from lung to the draining mediastinal lymph nodes (MLNs) is a key step for initiation of T cell responses in allergic asthma. In the present study, we investigate the role of DC-mediated airway inflammation after inhibition of p21-activated kinase-1 (PAK1), an effector of Rac and Cdc42 small GTPases, in the allergen-induced mouse models of asthma. Systemic administration of PAK1 specific inhibitor IPA-3 significantly attenuates not only the airway inflammation but also the airway hyperresponsiveness in a mouse model of ovalbumin-induced asthma. Specifically, intratracheal administration of low dosage of IPA-3 consistently decreases not only the airway inflammation but also the DC trafficking from lung to the MLNs. Importantly, intratracheal instillation of IPA-3-treated and ovalbumin-pulsed DCs behaves largely the same as that of either Rac inhibitor-treated and ovalbumin-pulsed DCs or Cdc42 inhibitor-treated and ovalbumin-pulsed DCs in attenuation of the airway inflammation in ovalbumin-challenged mice. Mechanistically, PAK1 is not involved in the maturation, apoptosis, antigen uptake, and T cell activation of cultured DCs, but PAK1 dose lie on the downstream of Rac and Cdc42 to regulate the DC migration toward the chemokine C-C motif chemokine ligand 19. Taken together, this study demonstrates that inhibition of PAK1 attenuates the cardinal features of asthma through suppressing the DC trafficking from lung to the MLN, and that interfere with DC trafficking by a PAK1 inhibitor thus holds great promise for the therapeutic intervention of allergic diseases.

Topics: Animals; Asthma; Bronchoconstrictor Agents; Cells, Cultured; Dendritic Cells; Disulfides; Dose-Response Relationship, Drug; Lymph Nodes; Mice; Mice, Inbred BALB C; Mice, Transgenic; Naphthols; Ovalbumin; p21-Activated Kinases

Nowadays, bronchial asthma is still a severe disease threatening human health, and it is incumbent upon us to seek effective therapeutic drugs. Mahuang decoction (MHD), a classic famous Chinese prescription, has been used for thousands of years to prevent phlegm from forming, stop coughing and relieve asthma, but the relevant mechanism has not been thoroughly clarified. This study aims to investigate the anti-airway inflammation effect of MHD and the possible molecular mechanism underlying IL21/STAT3 signaling pathway, so as to provide guidance for the treatment of MHD on bronchial asthma.. Specific pathogen free SD rats were randomly divided into 6 groups: normal control group, model group, positive group (Compound methoxyphenamine), MHD-treated groups at doses of 10 ml/kg, 5 ml/kg and 2.5 ml/kg, 10 rats in each group. Except for the normal control group, rats in other groups were sensitized with ovalbumin via introperitoneal injection and challenged with ovalbumin inhalation to trigger asthma model. At 24 h after the last excitation, bronchoalveolar lavage fluid (BALF) of every rat was drawn and the number of inflammatory cells was analyzed using cell counting method. ELISA method was performed to determine the concentrations of TXB. MHD intervention demonstrated a strong inhibitory action on the secretion of inflammatory mediators as well as the inflammatory cell infiltration in pulmonary tissues of asthmatic rats, and also depressed the protein expressions of IL-21, IL-21R, STAT3 and p-STAT3 in pulmonary tissues. MHD effectively mitigates airway inflammation and regulates the IL-21/STAT3 signaling pathway in rat asthma model.

Topics: 6-Ketoprostaglandin F1 alpha; Allergens; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Ephedra sinica; Leukocyte Count; Lung; Matrix Metalloproteinase 9; Ovalbumin; Phytotherapy; Plant Preparations; Rats, Sprague-Dawley; Signal Transduction; STAT3 Transcription Factor; Thromboxane B2; Tissue Inhibitor of Metalloproteinase-1

The mechanisms of allergen immunotherapy are not fully elucidated. Here, we sought to develop a murine model to demonstrate the effectiveness of subcutaneous immunotherapy (SCIT) for allergic responses. As excessive antigen dosages may induce immune tolerance in sensitized mice, the effects of SCIT were assessed by varying the antigen dosage. The mechanisms of SCIT were analyzed by focusing on the induction of Foxp3. The maximum effects of SCIT were observed with 1 mg/animal of OVA for airway inflammation induced by 5 μg/animal of OVA, in which airway eosinophilia and Th2 cytokine production were markedly suppressed. The increase in the OVA-specific IgE level was significantly suppressed by SCIT. The development of bronchial epithelial thickening and mucus accumulation were also suppressed by SCIT. Concomitantly, IL-10-producing Foxp3. We successfully developed an airway allergic model for SCIT. It was suggested that most of IL-10-producing Foxp3

Topics: Allergens; Animals; Asthma; Cells, Cultured; Cytokines; Desensitization, Immunologic; Disease Models, Animal; Eosinophils; Forkhead Transcription Factors; Humans; Hypersensitivity; Immune Tolerance; Infusions, Subcutaneous; Interleukin-10; Lung; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; T-Lymphocytes, Regulatory; Th1-Th2 Balance

The importance of developing new animal models to assess the pathogenesis of glucocorticoid (GC)-insensitive asthma has been stressed. Because of the asthma-prone background of A/J mice, we hypothesized that asthma changes in these animals would be or become resistant to GCs under repeated exposures to an allergen. A/J mice were challenged with OVA for 2 or 4 consecutive d, starting on day 19 postsensitization. Oral dexamethasone or inhaled budesonide were given 1 h before challenge, and analyses were done 24 h after the last challenge. Airway hyperreactivity, leukocyte infiltration, tissue remodeling, and cytokine levels as well as phosphorylated GC receptor (p-GCR), p-GATA-3, p-p38, MAPK phosphatase-1 (MKP-1), and GC-induced leucine zipper (GILZ) levels were assessed. A/J mice subjected to two daily consecutive challenges reacted with airway hyperreactivity, subepithelial fibrosis, and marked accumulation of eosinophils in both bronchoalveolar lavage fluid and peribronchial space, all of which were clearly sensitive to dexamethasone and budesonide. Conversely, under four provocations, most of these changes were steroid resistant. A significant reduction in p-GCR/GCR ratio following 4- but not 2-d treatment was observed, as compared with untreated positive control. Accordingly, steroid efficacy to transactivate MKP-1 and GILZ and to downregulate p-p38, p-GATA-3 as well as proinflammatory cytokine levels was also seen after two but not four provocations. In conclusion, we report that repeated allergen exposure causes GC-insensitive asthma in A/J mice in a mechanism associated with decrease in GCR availability and subsequent loss of steroid capacity to modulate pivotal regulatory proteins, such as GATA-3, p-p38, MKP-1, and GILZ.

Topics: Allergens; Animals; Asthma; Biological Availability; Bronchoalveolar Lavage Fluid; Budesonide; Cytokines; Dexamethasone; Disease Models, Animal; Down-Regulation; Eosinophils; Glucocorticoids; Hypersensitivity; Inflammation; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Glucocorticoid; Steroids; Transcriptional Activation

Toll-like receptor-4 (TLR4) is a key component of the innate immune system and activation of TLR4 signaling has a significant role in the pathogenesis of asthma. Therefore, our objective was to identify the natural TLR4 antagonist and evaluate its activity in experimentally induced asthma. Soya lecithin origin phosphatidylcholine (soya PC) was identified as a natural TLR4 antagonist by computational study. Based on the computational study, TLR4 antagonist activity of soya PC was confirmed in in vitro lipopolysaccharide (LPS)-induced neutrophil adhesion assay. In the in vivo study, rats were sensitized with ovalbumin (OVA) (100 μg/kg, i.p.) on the 7th, 14th and 21st days and challenged intranasally with OVA (100 μg/100 μL) and LPS (10 ng/100 μL), 4 days/wk for 3 weeks. At the end of the experiment, we performed lung function parameters (respiratory rate, tidal volume, airflow rate), inflammatory cytokines (interleukin [IL]-4, IL-5, IL-13), total and differential leukocytes in blood as well as bronchoalveolar lavage fluid (BALf) and histological examinations. The computational study indicated that TLR4 antagonist activity of soya PC is due to linoleic acid (18:2) fatty acid chain. Soya PC significantly suppressed the LPS-induced neutrophil adhesion in a concentration-dependent manner to 1 μg/mL. The treatment of soya PC (5 and 10 mg/kg, 18 days, i.p.) significantly improved the lung function parameters, total and differential leukocyte counts in blood and BALf in asthmatic rats. This efficacy of soya PC was in extent similar to dexamethasone (2.5 mg/kg, 18 days, i.p.). However, soya PC was superior to dexamethasone in terms of benefits. The protective action of soya PC may be due to TLR4 antagonist activity and linoleic acid composition.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Leukocyte Count; Lipopolysaccharides; Lung; Male; Models, Molecular; Ovalbumin; Phosphatidylcholines; Protein Conformation; Rats; Rats, Wistar; Toll-Like Receptor 4

Topics: Allergens; Animals; Asthma; Blood Platelets; Disease Models, Animal; Humans; Immunization; Mice; Ovalbumin; Platelet Activation

Allergic asthma is an inflammatory condition accompanied by inflammation as well as oxidative stress. Supplementation of an anti-inflammatory agent having antioxidant properties may have therapeutic effects against this disease. Over the recent decades, the interest in combination therapy as new alternative medication has increased and it offers numerous benefits along with noticeable lack of toxicity as well as side effects. In this study, protective effects of curcumin alone and in combination with piperine were evaluated in mouse model of allergic asthma. Balb/c mice were sensitized on days 0, 7, and 14 and challenged from days 16-30 on alternate days with ovalbumin (OVA). Mice were pretreated with curcumin (Cur; 10 and 20 mg/kg) and piperine (Pip; 5 mg/kg) alone and in combination via the intraperitoneal route on days 16-30 and compared with intranasal curcumin (5 mg/kg) treatment. Blood, bronchoalveolar lavage fluid (BALF), and lungs were collected after mice were sacrificed on day 31st. Mice immunized with OVA have shown significant increase in airway inflammation and oxidative stress as determined by oxidative stress markers. A significant suppression was observed with all the treatments, but intranasal curcumin treatment group has shown maximum suppression. So, among all the treatment strategies utilized, intranasal curcumin administration was most appropriate in reducing inflammation and oxidative stress and possesses therapeutic potential against allergic asthma. Present study may prove the possibility of development of curcumin nasal drops towards treatment of allergic asthma.

Topics: Alkaloids; Animals; Anti-Inflammatory Agents; Asthma; Benzodioxoles; Bronchoalveolar Lavage Fluid; Curcumin; Drug Administration Routes; Drug Therapy, Combination; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Piperidines; Polyunsaturated Alkamides

Asthma is a common airways inflammatory disease. This study provides evidence on the efficacy of flavocoxid against ovalbumin (OVA)-induced allergic airways inflammation in a mouse model of asthma. Airway inflammation was induced by intrapеritonеal injection of 10 mg ovalbumin (OVA) on day zero and day 7 followed by OVA challenge starting from 14th day to 16th day. Beclomethasone; a standard anti-inflammatory agent was selected as a drug in asthma. Flavocoxid (20 mg/kg, i. p.) was administered on day zero till 16th day followed by OVA challenge. At the end of the study, lung weight index, bronchoalveolar lavage fluid (BALF) content of total and differential WBCs, interleukin-13(IL-13), in addition to lung tissue nitrate/nitrite (NO) and oxidative stress biomarkers were measured. Also, histological and immunohistochemical analysis were conducted. Daily i. p. injection of flavocoxid (20 mg/kg) significantly improved airway inflammation. Inflammatory cells in BALF, malondialdehyde (MDA), NO and IL-13 significantly declined with concomitant increase in superoxide dismutase (SOD) activity. Histopathological examination and immunohistochеmical staining of mast cells were correlated with observed biochemical improvements. Collectively, these results demonstrate that flavocoxid mitigates the allergic airway inflammation induced by ovalbumin through attenuation of IL-13, NO expressions and oxidative stress.

Topics: Animals; Asthma; Biomarkers; Catechin; Disease Models, Animal; Drug Combinations; Inflammation; Lung; Mice; Ovalbumin; Oxidative Stress

We aimed to explore the imbalance between the T helper 17 γδT cells (γδT17) and the regulatory γδT cells (γδTreg) in asthmatic mice. Male Balb/c mice were randomly divided into the normal control group and the asthmatic model group. The asthmatic model group mice were intraperitoneally injected with the mixture of ovalbumin (OVA)/Al(OH)3 and then activated by exposure of the animals to OVA atomization. Airway hyperresponsiveness (AHR) was determined by a non-invasive lung function machine. Hematoxylin and eosin and Alcian blue-periodic acid Schiff staining were done for histopathological analysis. Interleukin (IL)-17 and IL-35 levels in bronchoalveolar lavage fluid were detected by ELISA. The percentage of IL-17+ γδT cells and Foxp3+ γδT cells in spleen cells suspension were detected and the transcription levels of RORγt and Foxp3 in the lung tissue were determined. Compared with the normal control, the severity of airway inflammation and AHR were higher in the asthmatic mice. Furthermore, mice in the asthmatic group displayed significant increases of IL-17+ γδT cells, expression of IL-17A, and RORγt, whereas control mice displayed marked decreases of Foxp3+ γδT cells, expression of IL-35, and transcription factor Foxp3. In addition, the mRNA expression of RORγt was positively correlated with the percentage of IL-17+γδT cells, and the mRNA level of Foxp3 was positively correlated with the percentage of Foxp3+ γδT cells. The imbalance of γδT17/γδTreg in the asthmatic mice may contribute to the pathogenesis of OVA-induced asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Interleukin-17; Interleukins; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Random Allocation; Real-Time Polymerase Chain Reaction; Th17 Cells

Mucus secretion accumulation in the airways may act as a contributing factor for the development of airflow limitation in severe fetal asthma patients. Accumulated evidences showed that transforming growth factor beta (TGF-β) plays a regulatory role in airway remodeling including mucus hyper-secretion in asthma. However, the detailed molecular mechanisms of TGF-β3 induced MUC5AC hyper-expression in airway epithelium remains unclear. Here, we demonstrated the pivotal roles of autophagy in regulation of MUC5AC hyper-production induced by TGF-β3 in airway epithelium. Our experimental data showed that inhibiting autophagy pathway in repeated ovalbumin (OVA) exposed mice exhibited decreased airway hyper-response and airway inflammation, diminishing the expression of Muc5ac and TGF-β3. Furthermore, our studies demonstrated that autophagy was induced upon exposure to TGF-β3 and then mediated MUC5AC hyper-expression by activating the activator protein-1 (AP-1) in human bronchial epithelial cells. Finally, Smad2/3 pathway was involved in TGF-β3-induced MUC5AC hyper-expressions by promoting autophagy. These data indicated that autophagy was required for TGF-β3 induced airway mucous hyper-production, and that inhibition of autophagy exerted therapeutic benefits for TGF-β3 induced airway mucus secretion.

Topics: Animals; Asthma; Autophagy; Bronchi; Disease Models, Animal; Epithelial Cells; Female; Humans; Mice; Mucin 5AC; Ovalbumin; Signal Transduction; Transcription Factor AP-1; Transforming Growth Factor beta3; Up-Regulation

Oxidative stress plays a key role in the immunopathogenesis of asthma. The objective of this study was to investigate the thymol effects on oxidative parameters along with trace elements in asthma experimental model. The Balb/c mice were sensitized by intraperitoneal injection of ovalbumin and thymol (8, 16 and 32 mg/kg) and dexamethasone (DEX) (2 mg/kg) were orally administered to sensitized mice. Oxidative stress parameters including protein carbonyl content, malondialdehyde (MDA), 8-hydroxy-2'-deoxyguanosine (8-OHdG) and total antioxidant capacity (TAC) besides trace element levels were evaluated. The protein carbonyl content, MDA and 8-OHdG in treated mice with 32 mg/kg of thymol significantly decreased compared to asthmatic mice (P < 0.01). Also, TAC significantly increased (P < 0.001) as well as zinc and selenium levels while copper level decreased. 16 mg/kg of thymol reduced the protein carbonyl content, MDA and 8-OHdG compared to asthmatic mice (P < 0.05). In addition, thymol improved the most prominent inflammation characteristics of asthma. The obtained results suggest that thymol has a protective effect against oxidative stress and it was also able to partially restore the defective trace element levels in asthma. Based on our observations, thymol may be used for alternative / complementary therapy in asthma.

Topics: Animals; Asthma; Disease Models, Animal; Female; Immunologic Factors; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidation-Reduction; Oxidative Stress; Thymol; Trace Elements; Treatment Outcome

Peroxisome proliferator-activated receptor-γ (PPARγ) agonist pioglitazone (PGZ) exhibits potential protective effects in asthma. Recently, regulator of G protein 4 (RGS4) has been reported to be associated with immunological and inflammatory responses. However, no evidence has shown the influence of PPARγ on RGS4 expression in airway disorders. In this study, BALB/c mice received ovalbumin (OVA) sensitization followed by OVA intranasal challenge for 90 days to establish a chronic asthma mouse model. Accompanied with OVA challenge, the mice received administration of PPARγ agonist PGZ (10 mg/kg) intragastrically or RGS4 inhibitor CCG 63802 (0.5 mg/kg) intratracheally. Invasive pulmonary function tests were performed 24 h after last challenge. Serum, bronchoalveolar lavage fluid (BALF), and lung tissues were collected for further analyses after the mice were sacrificed. We found that PPARγ agonist PGZ administration significantly attenuated the pathophysiological features of OVA-induced asthma and increased the expression of RGS4. In addition, the attenuating effect of PGZ on airway inflammation, hyperresponsiveness (AHR), and remodeling was partially abrogated by administration of RGS4 inhibitor CCG 63802. We also found that the downregulation of RGS4 by CCG 63802 also significantly increased inflammatory cell accumulation and AHR, and increased levels of IL-4, IL-13, eotaxin, IFN-γ, and IL-17A in BALF, and total and OV-specific IgE in serum. Furthermore, the inhibitory effects of PGZ on the activations of ERK and Akt/mTOR signaling, and MMPs were apparently reversed by CCG 63802 administration. In conclusion, the protective effect of PGZ on OVA-induced airway inflammation and remodeling might be partly regulated by RGS4 expression through ERK and Akt/mTOR signaling.

Topics: Airway Remodeling; Animals; Asthma; MAP Kinase Signaling System; Mice; Ovalbumin; Pioglitazone; PPAR gamma; Proto-Oncogene Proteins c-akt; RGS Proteins; Signal Transduction; TOR Serine-Threonine Kinases

MHTP [2-methoxy-4-(7-methoxy-1,2,3,4-tetrahydroisoquinolin-1-yl) phenol], a synthetic isoquinolinic alkaloid, presented anti-inflammatory activity in several experimental models of acute inflammation as lipopolysaccharide (LPS)-induced acute lung injury and phlogistic agent-induced edema and presented low preclinical toxicity. The aim of this study was to determine the MHTP effect on ovalbumin (OVA)-induced pulmonary allergic inflammation. In other to realize this study, female BALFB/c mice were sensitized and challenged with OVA (OVA group) and treated with MHTP (MHTP group) by nasal instillation. Inflammatory, allergic, and immunomodulatory parameters such as migration of inflammatory cells to the lung tissue, pulmonary histological analysis, serum level of IgE-allergen specific, cytokine secretion, and lung T cell population characterization were analyzed and the data were considered statistically significant with p < 0.05. OVA-sensitized and OVA-challenged and MHTP (5.0 mg/kg)-treated mice presented reduction on total leukocyte migration into the bronchoalveolar lavage (BALF) dependent of lymphocyte and eosinophil migration (p < 0.001 and p < 0.01) as compared with the OVA group. Flow cytometric analysis showed that MHTP treatment decreased the percentage of granulocytes (p < 0.001) into the BALF and lung tissue histological analyzes demonstrated that the MHTP treatment decreased leukocyte migration and mucus production. In addition, treatment with MHTP decreased the number of CD3

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Female; Granulocytes; Inflammation; Interferon-gamma; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes; Tetrahydroisoquinolines; Th1 Cells

This study was to investigate expression levels and functional activities of histone deacetylases (HDACs) with potential therapeutic targets selected in animal model of allergic asthma. Mice were sensitized and then challenged with saline (control) or ovalbumin (OVA) for 8 weeks. Airway resistance was determined by increasing concentrations of acetyl-β-methacholine chloride (0 - 50 mg/ml). The number of cells and cytokine production in bronchoalveolar lavage fluid (BALF) were determined by ELISA. Pathological changes of lung specimens were examined by histochemical staining methods under the light microscope. Expression and quantification of HDACs in lungs were measured using immunohistochemistry and Western blotting analysis. HDAC activity was identified using colorimetric and fluorometric methods. The OVA-treated mice had a significant enhancement in airway resistance with a large number of cells and increased interleukin (IL)-4 and -5 levels in BALF. Morphologically, an infiltration of inflammatory cells into epithelial layer with mucus accumulation and subepithelial fibrosis were seen in the OVA-exposed lungs. The expression levels for HDAC1, HDAC5, HDAC6, and HDAC8 were significantly elevated with weak induction of HDAC 2-4, which was identical with their catalytic activities detected in the lungs. In contrast, HDAC1 and HDAC5 activities were higher than others in the lungs. Individual HDACs are differently regulated in expression levels and functional activities in animal model of allergic asthma. Selective targeting of HDAC1/5 offers an opportunity to improve therapeutic effects of the disease.

Topics: Aerosols; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Female; Histone Deacetylases; Lung; Mice, Inbred BALB C; Ovalbumin

Dilodendron bipinnatum Radlk., Sapindaceae, a tree of the Mato Grosso Pantanal, is popularly known as "mulher-pobre". The decoction or infusion of its inner stem bark is used for treating inflammatory conditions.. To determine if a 70% hydroethanolic extract of Dilodendron bipinnatum stem bark (HEDb) is able to reduce allergic airway inflammation in a murine model of ovalbumin (OVA)-induced allergic asthma.. The inner stem bark powder was macerated in a 70% hydroethanolic solution (1:3 w/v) to obtain HEDb. The induction of experimental asthma was accomplished as follows: on days 1 and 10, Swiss mice were sensitized by an intraperitoneal injection of OVA (100 µg/mL) and aluminum hydroxide (10 µg/mL). From day 19 to 24, animals (n = 6/per group) were treated (p.o.) twice a day with either vehicle (distilled water), HEDb (20, 100 and 500 mg/kg) or dexamethasone (0.5 mg/kg). Sham group animals were intraperitoneally injected and challenged with saline solution (0.9%) instead of OVA and received distilled water orally instead of HEDb, whereas the other groups were challenged with OVA (3% in saline) by aerosolization. On day 25, bronchoalveolar lavage fluid (BALF) was collected for the quantification of total leukocytes, neutrophils, eosinophils, mononuclear cells and Th2 cytokines (IL-4, IL-5, and IL-13). The lungs were collected for histopathological analysis and blood was assayed to determine serum IgE levels. The anti-inflammatory activity of HEDb was additionally confirmed by a lipoxygenase (LO) inhibitory assay in vitro.. These results somewhat agree on the popular use of the inner stem bark of D. bipinnatum as a treatment for allergic asthma. The HEDb exhibits significant anti-inflammatory activity in the OVA-induced mouse model of allergic asthma, possibly due to the down-regulation of the Th2 responses and LO inhibition, resulting in improvements in all analyzed inflammatory parameters.

Topics: Allergens; Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Immunoglobulin E; Leukocyte Count; Lipoxygenase Inhibitors; Male; Mice; Ovalbumin; Plant Bark; Sapindaceae

Diisononyl phthalate (DINP) and formaldehyde both are associated with asthma and allergies. However, it is unclear about the adverse effect of DINP and formaldehyde exposure on the brain for asthma patients. Here, we determined the effect of DINP and/or formaldehyde exposure on neuroinflammation in brain by a murine asthma model and investigated the underlying mechanisms. Mice were exposed to formaldehyde and/or DINP and sensitization with ovalbumin. The results show that exposure to formaldehyde and/or DINP not only exacerbated allergic asthma-like symptoms, but also promoted neuroinflammation in brain. The incrassation of the airway wall and exacerbation of neuroinflammation were more obviously when mice were subjected to a combined exposure to DINP and formaldehyde. Exposure to DINP and/or formaldehyde enhances oxidative stress and the activation of NF-κB in the prefrontal cortex of mouse asthma model. Exposure to DINP and/or formaldehyde also induced an increase in IL-1β, IL-17, and NGF. Blocking oxidative stress by administering melatonin or inhibiting NF-κB activation by treatment with Dehydroxymethylepoxyquinomicin effectively prevented increasing the levels IL-1β, IL-17 and nerve growth factor. The data indicated that DINP and/or formaldehyde exposure promoted neuroinflammation in the brain through enhanced oxidative stress and activation of NF-κB in a mouse asthma model.

Topics: Animals; Asthma; Disease Models, Animal; Encephalitis; Formaldehyde; Gene Expression Regulation; Interleukin-17; Interleukin-1beta; Male; Mice; Mice, Inbred BALB C; Nerve Growth Factor; NF-kappa B; Ovalbumin; Oxidative Stress; Phthalic Acids; Prefrontal Cortex; Respiratory Hypersensitivity; Signal Transduction

Asthma is the most common chronic childhood disease worldwide, characterized by airway remodeling and chronic inflammation, orchestrated primarily by Th2 cytokines. The aim of the current study was to explore the influences of milk fat globule epidermal growth factor 8 (MFG-E8)/integrin β3 signaling involved in airway inflammation and remodeling in asthma. BALB/c mice were sensitized by intraperitoneal injection of ovalbumin (OVA), followed by OVA nebulization. The levels of MFG-E8 expression were declined markedly in the OVA-induced allergy murine model. In addition, administration of MFG-E8 strongly reduced the accumulation of T-helper type 2 (Th2)-associated cytokines (such as interleukin-4, -5, and -13) as well as chemokine CCL11 (eotaxin) in bronchoalveolar lavage fluid and tissues in the OVA-sensitized mice. Moreover, MFG-E8 remarkably repressed the total immunoglobulin E and OVA-specific immunoglobulin E in serum in OVA-challenged mice. Meanwhile, treatment with recombinant murine MFG-E8 noticeably prevented inflammatory cell infiltration into the airways, as showed by a marked decrease in the numbers of total immune cells, eosinophils, neutrophils, macrophages, and lymphocytes in the bronchoalveolar lavage fluid in response to OVA challenge. Importantly, MFG-E8 apparently alleviated OVA-driven airway remodeling, which were evidenced by declined secretion of important mediators of airway remodeling, including transforming growth factor-β1, matrix metalloproteinase 9, ADAM8, and vascular endothelial growth factor, and reduced airway collagen deposition and inhibited goblet cell hyperplasia in OVA-induced asthma in mice. Mechanistically, integrin 3 contributes to the protective effect of MFG-E8 in inhibiting airway inflammation and remodeling in OVA-driven features of allergic asthma. Overall, MFG-E8, as a candidate molecule to evaluate airway inflammation and remodeling, could be a potential target for the management and prevention of asthma exacerbations, suggesting that MFG-E8/integrin β3 signaling may serve as a promising therapeutic agent for childhood asthma.

Topics: Airway Remodeling; Analysis of Variance; Animals; Antigens, Surface; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Gene Knockdown Techniques; Immunoglobulin E; Inflammation; Integrin beta3; Mice; Mice, Inbred BALB C; Milk Proteins; Ovalbumin; Recombinant Proteins; RNA, Messenger; Transforming Growth Factor beta1

Topics: Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Female; Furocoumarins; Kv1.3 Potassium Channel; Lipopolysaccharides; MAP Kinase Signaling System; Mice, Inbred BALB C; NF-kappa B; Ovalbumin

Achieving the optimal clinical outcome of mesenchymal stem cells (MSCs) is particularly dependent on fundamental understanding of therapeutic mechanisms. The current study was focused on the possible mechanisms by which rat bone marrow-derived mesenchymal stem cells (rBMMSCs) and/or conditioned media (CM) display broad immunomodulatory properties for ameliorating of asthma-related pathological changes. Male rats were divided equally into four experimental groups (n = 6): healthy rats received 50 μl PBS intravenously (group C), sensitized rats received 50 μl PBS intravenously (group OVA), sensitized rats received 50 μl CM intravenously (group OVA + CM), and sensitized rats received 50 μl PBS intravenously containing 2 × 10

Topics: Animals; Asthma; Cell Adhesion Molecules; Culture Media, Conditioned; Endothelial Cells; Intercellular Adhesion Molecule-1; Interferon-gamma; Interleukin-12; Interleukin-5; Lung; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Ovalbumin; Rats; Vascular Cell Adhesion Molecule-1

Adipose-derived mesenchymal stem cell (ASCs) exerts immunomodulatory roles in asthma. However, the underlying mechanism remains unclear. The present study aimed to explore the effects and mechanisms of ASCs on chronic asthma using an ovalbumin (OVA)-sensitized asthmatic mouse model.. Murine ASCs (mASCs) were isolated from male Balb/c mice and identified by the expression of surface markers using flow cytometry. The OVA-sensitized asthmatic mouse model was established and then animals were treated with the mASCs through intratracheal delivery. The therapy effects were assessed by measuring airway responsiveness, performing immuohistochemical analysis, and examining bronchoalveolar lavage fluid (BALF). Additionally, the expression of inflammatory cytokines and lgE was detected by CHIP and ELISA, respectively. The mRNA levels of serum indices were detected using qRT-PCR.. The mASCs grew by adherence with fibroblast-like morphology, and showed the positive expression of CD90, CD44, and CD29 as well as the negative expression of CD45 and CD34, indicating that the mASCs were successfully isolated. Administering mASCs to asthmatic model animals through intratracheal delivery reduced airway responsiveness, the number of lymphocytes (P < 0.01) and the expression of lgE (P < 0.01), IL-1β (P < 0.05), IL-4 (P < 0.001), and IL-17F (P < 0.001), as well as increased the serum levels of IL-10 and Foxp3, and the percentage of CD4 + CD25 + Foxp3+ Tregs in the spleen, and reduced the expression of IL-17 (P < 0.05) and RORγ.. Intratracheal administration of mASCs alleviated airway inflammation, improved airway remodeling, and relieved airway hyperresponsiveness in an OVA-sensitized asthma model, which might be associated with the restoration of Th1/Th2 cell balance by mASCs.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Immunoglobulin E; Lung; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes, Regulatory; Th1-Th2 Balance

T helper 17 (Th17) cells has proven to be crucial in the pathogenesis of severe asthma. Although it is known that Suppressor of cytokine signaling 3 (SOCS3) is involved in differentiation of Th17 cells but, how it affects severe asthma is uncertain. Since previous studies indicated that Methtyl-CpG binding domain protein 2 (MBD2) null mice was deficient in Th17 cell differentiation, the aim of the present study was to understand how MBD2 interacts with SOCS3 to regulate Th17 cell differentiation in severe asthma. Here, we show that SOCS3 expression was significantly decreased in Th17-mediated severe asthmatic mice, accompanied by elevated STAT3 phosphorylation and RORγt expression. Knock-down of SOCS3 promoted the differentiation of naïve T cells into Th17 cells through STAT3/RORγt pathway. Meanwhile, MBD2 was overexpressed in Th17-mediated severe asthmatic mice. Intervention of MBD2 expression lead to a negative change of SOCS3 expression, whereas the differentiation of Th17 cells showed positive change. In addition, MBD2 knockout (MBD2-KO) mice displayed increased SOCS3 expression and decreased Th17 differentiation after severe asthma modeling. Taken together, our results suggest that MBD2 might facilitate Th17 cell differentiation via down-regulating SOCS3 expression in severe asthma. These findings uncover new roles for SOCS3 and MBD2, and provide a potential target for treatment of severe asthma.

Topics: Allergens; Animals; Asthma; Cell Differentiation; Disease Models, Animal; DNA-Binding Proteins; Female; Gene Expression Regulation; Humans; Lipopolysaccharides; Lung; Mice; Mice, Inbred C57BL; Mice, Knockout; Nuclear Receptor Subfamily 1, Group F, Member 3; Ovalbumin; Phosphorylation; Pyroglyphidae; Severity of Illness Index; Signal Transduction; STAT3 Transcription Factor; Suppressor of Cytokine Signaling 3 Protein; Th17 Cells

Chronic helminth infection with Schistosoma (S.) mansoni protects against allergic airway inflammation (AAI) in mice and is associated with reduced Th2 responses to inhaled allergens in humans, despite the presence of schistosome-specific Th2 immunity. Schistosome eggs strongly induce type 2 immunity and allow to study the dynamics of Th2 versus regulatory responses in the absence of worms. Treatment with isolated S. mansoni eggs by i.p. injection prior to induction of AAI to ovalbumin (OVA)/alum led to significantly reduced AAI as assessed by less BAL and lung eosinophilia, less cellular influx into lung tissue, less OVA-specific Th2 cytokines in lungs and lung-draining mediastinal lymph nodes and less circulating allergen-specific IgG1 and IgE antibodies. While OVA-specific Th2 responses were inhibited, treatment induced a strong systemic Th2 response to the eggs. The protective effect of S. mansoni eggs was unaltered in μMT mice lacking mature (B2) B cells and unaffected by Treg cell depletion using anti-CD25 blocking antibodies during egg treatment and allergic sensitization. Notably, prophylactic egg treatment resulted in a reduced influx of pro-inflammatory, monocyte-derived dendritic cells into lung tissue of allergic mice following challenge. Altogether, S. mansoni eggs can protect against the development of AAI, despite strong egg-specific Th2 responses.

Topics: Allergens; Alum Compounds; Animals; Antibodies, Protozoan; Asthma; Cytokines; Disease Models, Animal; Eosinophilia; Female; Immunoglobulin E; Immunoglobulin G; Inflammation; Interleukin-2 Receptor alpha Subunit; Lung; Mice, Inbred C57BL; Ovalbumin; Ovum; Schistosoma mansoni; T-Lymphocytes, Regulatory; Th2 Cells

Curcumin has shown various pharmacological effects such as anti-inflammatory activities. In this study, the effects of curcumin on tracheal responsiveness and lung pathological features were evaluated in a rat model of asthma.. Tracheal responsiveness and lung pathological features were evaluated in control rats (C), ovalbumin (OVA)-sensitized rats (as an animal model of asthma; A), A rats treated with curcumin (Cu, 0.15, 0.30, and 0.60mg/ml) and dexamethasone (D, 1.25μg/ml), (n=8 in curcumin-treated groups and n=6 in other groups). Curcumin and dexamethasone were added to animals' drinking water during the sensitization period.. Asthmatic group showed increased lung pathological score and tracheal responsiveness to methacholine and OVA compared to control group (p<0.01 to p<0.001). Pathological features including interstitial inflammation, interstitial fibrosis, bleeding, and emphysema as well as tracheal responsiveness to methacholine and OVA, were significantly decreased in treated groups with dexamethasone and all concentrations of curcumin compared to group A (p<0.05 to p<0.001). Epithelial damage was also significantly decreased in treated groups with the two higher concentrations of curcumin (p<0.05 to p<0.001).. Curcumin showed preventive effects on tracheal responsiveness and lung pathological features in asthmatic rats.

Topics: Animals; Asthma; Curcumin; Dexamethasone; Dose-Response Relationship, Drug; Isometric Contraction; Lung; Male; Methacholine Chloride; Ovalbumin; Rats; Trachea

Nicotine, an nAChR agonist, shows prominent anti-inflammatory properties, and some studies have illustrated its suppressive effects on inflammation. Here, we have examined whether nicotine as a medicine may have beneficial effects on the treatment of asthma in a mouse model of allergic asthma. BALB/c mice were sensitized with OVA and alum. Two weeks later, the mice received nicotine with concentrations of 1 and 10 mg/kg three times every other day. After 10 days, the mice were challenged with OVA (5%) using an ultrasonic nebulizer and died the next day. Our results showed that the administration of nicotine reduced lung-tissue inflammation, the number of eosinophils in bronchoalveolar fluid, allergen-specific IgE and IL-4 production, while it increased the TGF-β/IL-4 ratio and the number of Treg cells. Our results showed that nicotine applies its suppressive effects in a dose-dependent manner: administration of 10 mg/kg of nicotine showed more suppressive effects than 1 mg/kg. Such data suggested that nicotine might be a good candidate to be used as a medicine in the treatment of allergic asthma by decreasing allergic inflammation severity and potentiating Treg cells proliferation against the allergen.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Eosinophils; Immunoglobulin E; Interleukin-4; Male; Mice, Inbred BALB C; Nicotine; Ovalbumin; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

To explore the therapeutic effect of honokiol on particulate matter 2.5 (PM2.5)-induced lung injury in asthmatic mice and the possible mechanisms.
 Methods: A total of 32 BALB/C mice were randomly divided into four groups: a normal saline group, a model group, a PM2.5 group and a honokiol group (n=8 in each group). The asthma mouse model was established by ovalbumin treatment. The mice were treated with physiological saline, ovalbumin, PM2.5 and honokiol, respectively. Lung tissues and serum were collected. The pathological changes of lung tissues were evaluated. The levels of inflammatory cytokines in bronchoalveolar lavage fluid (BALF) and serum were measured and the expressions of Toll like receptor 4 (TLR4), nuclear factor kappa B (NF-κB), retinoid-related orphan receptor gamma-t (RORγt) and forkhead box protein 3 (Foxp3) in lung tissues were detected.
 Results: 1) The lung tissues of mice in the asthma group showed obvious pathological changes and inflammatory state, suggesting that the asthma model was established successfully. PM2.5 could aggravate the pathological condition of inflammatory injury in lung tissues in asthmatic mice. 2) Compared to the PM2.5 group, the pathological symptoms in the lung tissues were alleviated in the honokiol group and the percentage of inflammatory cells in BALF and the levels of inflammatory cytokines in BALF and serum were significantly reduced (all P<0.05). 3) Compared to the PM2.5 group, the expressions of TLR4, NF-κB (p-p65) and RORγt in lung tissues were significantly decreased, while the expression of Foxp3 was increased; the ratio of RORγt/Foxp3 was also decreased in the honokiol group (all P<0.05).
 Conclusion: Honokiol can resist lung injury induced by PM2.5 in asthmatic mice. These effects are through inhibiting TLR4-NF-κB pathway-mediated inflammatory response or regulating the balance of Th17/Treg cells.. 目的：探究和厚朴酚对颗粒物2.5(particulate matter 2.5，PM2.5)诱导的哮喘小鼠肺损伤的治疗作用及其可能的作用机制。方法：32只BALB/C小鼠随机分为生理盐水组、模型组、PM2.5组和厚朴酚组，每组8只。使用卵清蛋白诱导哮喘小鼠模型，分别采用生理盐水、卵清蛋白、PM2.5和和厚朴酚处理，收集各组小鼠肺组织和血清，检测小鼠肺组织的病理损伤状态、支气管肺泡灌洗液(bronchoalveolar lavage fluid，BALF)和血清中炎性因子的表达水平，以及肺组织中Toll样受体4(Toll like receptor 4，TLR4)、核因子κB(nuclear factor kappa B，NF-κB)、维甲酸相关核孤儿受体γt(retinoid-related orphan receptor gamma-t，RORγt)和叉头状转录蛋白3(forkhead box protein 3，Foxp3)的蛋白表达水平。结果：1)模型组小鼠肺组织出现明显病理损伤和炎性状态，提示哮喘模型构建成功。PM2.5能够加重哮喘小鼠的肺组织病理损伤和炎性状态；2)与PM2.5组比较，和厚朴酚组小鼠肺组织的病理损伤状态得到缓解，BALF中炎性细胞减少，炎性因子水平降低(均P<0.05)。3)与PM2.5组比较，和厚朴酚组小鼠肺组织中TLR4，NF-κB(p-p65)和RORγt的表达减少，Foxp3表达水平增加，且RORγt/Foxp3比值减少(均P<0.05)。结论：和厚朴酚能够抵抗PM2.5诱导哮喘小鼠的肺损伤，这一方面可能是通过抑制TLR4-NF-κB信号通路介导的炎症反应，另一方面可能是通过影响T辅助细胞17/调节性T细胞平衡的方式来实现的。.

Topics: Animals; Asthma; Biphenyl Compounds; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Inflammation Mediators; Lignans; Lung; Lung Injury; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Particulate Matter; Random Allocation; Toll-Like Receptor 4

Asthma is a syndrome with multifactorial causes, resulting in a variety of different phenotypes. Current treatment options are not curative and are sometimes ineffective in certain disease phenotypes. Therefore, novel therapeutic approaches are required. Recent findings have shown that activation of the canonical Wnt signaling pathway suppresses the development of allergic airway disease. In contrast, the effect of the noncanonical Wnt signaling pathway activation on allergic airway disease is not well described. The aim of this study was to validate the therapeutic effectiveness of Wnt-1-driven canonical Wnt signaling compared with Wnt-5a-driven noncanonical signaling in murine models. In vitro, both ligands were capable of attenuating allergen-specific T cell activation in a dendritic cell-dependent manner. In addition, the therapeutic effects of Wnt ligands were assessed in two different models of allergic airway disease. Application of Wnt-1 resulted in suppression of airway inflammation as well as airway hyperresponsiveness and mucus production. In contrast, administration of Wnt-5a was less effective in reducing airway inflammation or goblet cell metaplasia. These results suggest an immune modulating function for canonical as well as noncanonical Wnt signaling, but canonical Wnt pathway activation appears to be more effective in suppressing allergic airway disease than noncanonical Wnt activation.

Topics: Allergens; Animals; Asthma; Cells, Cultured; Dendritic Cells; Disease Models, Animal; Female; Humans; Immunomodulation; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Respiratory Hypersensitivity; T-Lymphocytes; Wnt Signaling Pathway; Wnt-5a Protein; Wnt1 Protein

Depression is very common in asthmatic patients and may increases risk for morbidity and mortality. The present work aimed to investigate the protective effect of fluoxetine, on behavioral and biochemical changes, associated with ovalbumin (OVA) - induced bronchial asthma and depression in rats. Rats were sensitized with intraperitoneal administration of OVA plus aluminum hydroxide for 3 consecutive days then at day 11 followed by OVA intranasal challenge at days 19, 20, 21. Rats were either pretreated with dexamethasone, fluoxetine10mg/kg or fluoxetine 20 mg/kg. At the end of the experiment, various tests were performed, including open field, forced swimming and respiratory function tests. Blood was drawn for serum IgE detection. Finally, rats were euthanized, brain-derived neurotrophic factor (BDNF) was estimated in bronchoalveolar lavage (BAL) fluid and lung content of reduced glutathione (GSH), superoxide dismutase (SOD), tumor necrosis factor-alpha (TNF-α) and interleukin 4 (IL-4) were determined. Histopathological study was also performed. The results showed that fluoxetine significantly ameliorated OVA- induced biochemical and behavioral changes. Fluoxetine may protect against OVA-induced asthma and depression in rats. This effect may be mediated at least in part by its antioxidant, anti-inflammatory and immunosuppressant effect.

Topics: Animals; Asthma; Brain-Derived Neurotrophic Factor; Depression; Dexamethasone; Fluoxetine; Immunoglobulin E; Interleukin-4; Male; Ovalbumin; Oxidative Stress; Peak Expiratory Flow Rate; Rats

Perturbation of commensal bacteria by antibiotic exposure aggravates ovalbumin (OVA)-induced allergic asthma in pre-weaning mice. However, the influence of dysbiosis of commensal bacteria on asthma development in post-weaning mice is still limited. Here, we treated 3-week-old post-weaning mice with antibiotics to disrupt commensal bacteria and then established OVA-induced allergic asthma by peritoneal sensitization using OVA/alum and intranasal challenge with OVA. Contrary to the protective function in pre-weaning mice, commensal bacteria in post-weaning mice aggravated OVA-induced asthma. Commensal bacteria in post-weaning mice promoted OVA-induced allergic asthma through maintenance of NLRP3/IL-1β expression in peritoneal macrophages (pMφ), which promoted recruitment of inflammatory cells, especially inflammatory monocytes, into the peritoneal cavity after OVA/alum sensitization. Further study showed that metronidazole- and vancomycin-sensitive bacteria are involved in maintenance of NLRP3/IL-1β signal in pMφ. Our results suggest that certain species of commensal bacteria in post-weaning mice aggravate OVA-induced allergic asthma through NLRP3/IL-1β signal pathway.

Topics: Animals; Animals, Newborn; Anti-Bacterial Agents; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Epithelial Cells; Gene Expression; Interleukin-1beta; Lung; Macrophages, Peritoneal; Metronidazole; Mice; Mice, Inbred C57BL; Mice, Knockout; Microbiota; NLR Family, Pyrin Domain-Containing 3 Protein; Ovalbumin; Signal Transduction; Symbiosis; Vancomycin; Weaning

Augmented smooth muscle contractility of the airways is one of the causes of airway hyperresponsiveness in asthmatics. However, the mechanism of the altered properties of airway smooth muscle cells is not well understood.. To identify differentially expressed genes (DEGs) related to the bronchial smooth muscle (BSM) hyper-contractility in a murine asthma model.. The ovalbumin (OA)-sensitized mice were repeatedly challenged with aerosolized OA to induce asthmatic reaction. Transcriptomic profiles were generated by microarray analysis of BSM tissues from the OA-challenged and control animals, and KEGG (Kyoto Encyclopedia of Genes and Genomes) Pathway Analysis was applied.. Tension study showed a BSM hyperresponsiveness to acetylcholine (ACh) in the OA-challenged mice. A total of 770 genes were differentially expressed between the OA-challenged and control animals. Pathway analysis showed a significant change in arachidonic acid (AA) metabolism pathway in BSM tissues of the OA-challenged mice. Validation of DEGs by quantitative RT-PCR showed a significant increase in PLA2 group 4c (Pla2g4c)/COX-2 (Ptgs2)/PGD2 synthase 2 (Hpgds) axis. PGD2 level in bronchoalveolar fluids of the OA-challenged mice was significantly increased. A 24-h incubation of BSM tissues with PGD2 caused a hyperresponsiveness to ACh in naive control mice.. AA metabolism is shifted towards PGD2 production in BSM tissues of asthma. Increased PGD2 level in the airways might be a cause of the BSM hyperresponsiveness in asthma.

Topics: Acetylcholine; Animals; Asthma; Bronchi; Cyclooxygenase 2; Disease Models, Animal; Gene Expression Regulation; Group IV Phospholipases A2; Humans; Intramolecular Oxidoreductases; Mice; Mice, Inbred BALB C; Muscle Contraction; Muscle, Smooth; Ovalbumin; Respiratory Hypersensitivity

The efficacy of heparins and low-MW-heparins (LMWH) against human asthma has been known for decades. However, the clinical utility of these compounds has been hampered by their anticoagulant properties. Much effort has been put into harnessing the anti-inflammatory properties of LMWH but none have been used as therapy for asthma. Sulfated-non-anticoagulant heparin (S-NACH) is an ultra-LMWH with no systemic anticoagulant effects.. The present study explored the potential of S-NACH in blocking allergic asthma and examined the potential mechanism by which it exerts its effects.. Acute and chronic ovalbumin-based mouse models of asthma, splenocytes, and a lung epithelial cell line were used. Mice were challenged with aerosolized ovalbumin and administered S-NACH or saline 30 min after each ovalbumin challenge.. Sulfated-non-anticoagulant heparin administration in mice promoted a robust reduction in airway eosinophilia, mucus production, and airway hyperresponsiveness even after chronic repeated challenges with ovalbumin. Such effects were linked to suppression of Th2 cytokines IL-4/IL-5/IL-13/GM-CSF and ovalbumin-specific IgE without any effect on IFN-γ. S-NACH also reduced lung fibrosis in mice that were chronically-exposed to ovalbumin. These protective effects of S-NACH may be attributed to modulation of the IL-4/JAK1 signal transduction pathway through an inhibition of STAT6 phosphorylation and a subsequent inhibition of GATA-3 and inducible NO synthase expression. The effect of the drug on STAT6 phosphorylation coincided with a reduction in JAK1 phosphorylation upon IL-4 treatment. The protective effects of S-NACH treatment was associated with reduction of the basal expression of the two isoforms of arginase ARG1 and ARG2 in lung epithelial cells.. Our study demonstrates that S-NACH constitutes an opportunity to benefit from the well-known anti-asthma properties of heparins/LMWH while bypassing the risk of bleeding. Our results show, for the first time, that such anti-asthma effects may be associated with reduction of the IL-4/JAK1/STAT6 pathway.

Topics: A549 Cells; Animals; Anti-Inflammatory Agents; Anticoagulants; Asthma; Cell Line; Cell Separation; Disease Models, Animal; Flow Cytometry; Hemorrhage; Heparin; Humans; Hypersensitivity; Interleukin-4; Janus Kinase 1; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Signal Transduction; Spleen; STAT6 Transcription Factor; Th2 Cells

Annexin A5 (ANXA5) has a potential role in cellular signal transduction, inflammation, and fibrosis. However, the exact role of ANXA5 in asthma remains to be clarified. The aims of the present study were to investigate ANXA5 protein expression in a mouse model of asthma and pollutant exposure and to elucidate the relationships between clinical variables and plasma ANXA5 levels in patients with asthma.. ANXA5 protein levels were lower in lung tissue from OVA + OVA mice than in control mice. Lung ANXA5, connective tissue growth factor (CTGF), and transforming growth factor β1 (TGF-β1) protein levels were higher in OVA + TiO. Our results imply that ANXA5 plays a potential role in asthma pathogenesis and may be a promising marker for exacerbated bronchial asthma and exposure to air pollutants.

Topics: A549 Cells; Aged; Air Pollutants; Animals; Annexin A5; Antigens, Dermatophagoides; Asthma; Biomarkers; Connective Tissue Growth Factor; Dermatophagoides pteronyssinus; Disease Models, Animal; Disease Progression; Female; Forced Expiratory Volume; Humans; Male; Mice; Mice, Inbred BALB C; Middle Aged; Nanoparticles; Ovalbumin; Pulmonary Fibrosis; Titanium; Transforming Growth Factor beta1; Vital Capacity

Epithelial cells (ECs)-derived cytokines are induced by different stimuli through pattern recognition receptors (PRRs) to mount a type-2-cell-mediated immune response; however, the underlying mechanisms are poorly characterized. Here, we demonstrated asthmatic features in both primary bronchial epithelial cells (pBECs) and mouse model using several allergens including ovalbumin (OVA), house dust mite (HDM), or Alternaria alternata. We found that toll-like receptor 2 (TLR2) was highly induced in ECs but not dendritic cells (DCs) by various allergens, leading to recruitment of circulating basophils into the lung via C-C chemokine ligand-2 (CCL2). TLR2 expression increased thymic stromal lymphopoietin (TSLP) production through the NF-κB and JNK signaling pathways to extend the survival of recruited basophils and resident DCs in the lung, predisposing a type-2-cell-mediated airway inflammation. Conversely, TLR2 deficiency impaired secretion of TSLP and CCL2, decreased infiltration of lung basophils, and increased resistance to Th2 response. Blocking TSLP also phenocopied these phenomena. Our findings reveal a pro-inflammatory role of airway ECs through a TLR2-dependent TSLP production, which may have implication for treating allergic asthma.

Topics: Allergens; Alternaria; Animals; Asthma; Basophils; Bronchi; Cells, Cultured; Chemokine CCL2; Cytokines; Dendritic Cells; Disease Models, Animal; Epithelial Cells; Inflammation; Lung; Mice; Mice, Inbred C57BL; Ovalbumin; Pyroglyphidae; Th2 Cells; Thymic Stromal Lymphopoietin; Toll-Like Receptor 2

Asthma is an inflammatory disease characterized by airway hyperresponsiveness. Gallic acid is a powerful anti-inflammatory agent. In this study we aimed to investigate the efficacy of gallic acid in asthma treatment and its mechanisms.. An ovalbumin-induced asthma mouse model was generated. Pro-inflammatory cell infiltration and T helper (Th2)-associated cytokine release in the bronchoalveolar lavage fluid (BALF) were analyzed to reflect the severity of asthma in mice. An interleukin-33 (IL-33)-induced asthma mouse model was also generated to study the mechanism by which gallic acid could improve asthma. Group 2 lymphoid cells (ILC2s) were identified using flow cytometry. Proteins were detected using Western blotting.. Ovalbumin significantly increased the infiltration of pro-inflammatory cells, including eosinophils, macrophages, lymphocytes, and neutrophils, accompanied by enhanced airway hyperesponsiveness. Gallic acid reduced pro-inflammatory cell infiltration and improved airway hyperresponsiveness. Meanwhile, gallic acid reduced IL-5 and IL-13 levels in BALF and decreased expression of IL-33 in the lungs. Mechanistically, gallic acid inhibited MyD88 expression and downregulated nuclear factor (NF)-κB signaling to decrease IL-33 expression.. Gallic acid can mollify ovalbumin-induced asthma in mice, possibly by inhibiting IL-33-mediated ILC2 activation and subsequent Th2 cytokine release via downregulation of the MyD88/NF-κB signaling pathway. ©2018 ARSAAOA, LLC.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Female; Gallic Acid; Immunity, Innate; Interleukin-33; Lymphocytes; Mice, Inbred BALB C; Ovalbumin

Topics: Animals; Anti-Allergic Agents; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchi; Calcium Channel Blockers; Cytokines; Gabapentin; Lung; Male; Mice; Ovalbumin

Oxidative stress induced by cigarette smoke and other environmental pollutants contributes to refractory asthma. To better understand the role of smoking in asthma, we investigated the effects of cigarette smoke on allergic airway responses in mice and examined expression of nuclear factor-E2-related factor-2 (Nrf2) and its downstream factors, because Nrf2 is known to play a pivotal role in antioxidant responses. OVA-sensitized and challenged BALB/c mice were exposed to cigarette smoke and then treated with dexamethasone, sulforaphane (an activator of Nrf2), or their combination. Upon exposure to cigarette smoke, Nrf2 and associated transcripts were upregulated in response to oxidative stress, and asthmatic responses were steroid resistant. In OVA-sensitized and challenged mice exposed to cigarette smoke and treated with sulforaphane, Nrf2-mediated antioxidant responses were upregulated to a greater extent, and steroid sensitivity of asthmatic responses was restored. Moreover, the expression and activity of histone deacetylase 2 (HDAC2), a key regulator of steroid responsiveness, was reduced in mice exposed to cigarette smoke, but restored by sulforaphane treatment. No effects of sulforaphane were observed in Nrf2-deficient mice. These findings indicate that cigarette smoke induces steroid unresponsiveness in asthmatic airways, and that sulforaphane restores steroid sensitivity via upregulation of Nrf2 and enhancement of HDAC2 expression and activity. Thus, Nrf2 may serve as a potential molecular target for cigarette smoke-related refractory asthma resistant to steroid therapy.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Dexamethasone; Disease Models, Animal; Drug Combinations; Female; Gene Expression Regulation; Histone Deacetylase 2; Isothiocyanates; Lung; Mice; Mice, Inbred BALB C; Mice, Knockout; Nicotiana; Nuclear Respiratory Factor 1; Ovalbumin; Oxidative Stress; RNA, Messenger; Signal Transduction; Sulfoxides; Tobacco Smoke Pollution

To examine the effect of metanol extract of Petiveria alliacea (PM) on airway inflflammation in a murine model of chronic asthma.. Two-month-old male BALB/c mice (n=6-8/group) were sensitized on days 0 and 14 by intraperitoneal injection of 20 μg ovalbumin (OVA). On day 25, the mice received an airway challenge with OVA (3%, w/v, in phosphate buffered saline). PM was administered orally by oral gavage to mice at doses of 100, 200 and 400 mg/kg body weight once daily from days 18 to 23. Control mice were orally administered phosphate buffered saline (PBS) to induce a model of asthma. At the end of the test, respiratory reactivity was assayed, the total cell number, interleukin-4 (IL-4), IL-5, IL-13, tumor necrosis factor-alpha (TNF-α) and reactive oxygen species (ROS) in the bronchoalveolar lavage fluid (BALF) were determined and the levels of serum IgE, intercellular cell adhesion molecule 1 (ICAM-1) and eotoxin were measured. In addition, lung tissue was used to qualify the IL-4, IL-5, IL-13, TNF-α and transforming growth factor beta 1 (TGF-β1). Histologic examination was performed to observe inflammatory cellular infiltration.. The administration of PM in comparison with the OVA-only treated group signifificantly attenuated the infifiltration of eosinophils and other inflflammatory cells (P<0.01). Airway resistance (RI) in the OVA-only induced group was significantly higher than that of the PBS control group (P<0.01) when methacholine was added. TNF-α, IgE, TGF-β1 and cytokine levels IL-4, IL-5, IL-13 in the BALF decreased compared to control mice (P<0.01 or P<0.05). PM treatment also inhibited the production of chemokines, eotaxin and ICAM-1 in BALF (P<0.01), which improved lung function. Histopathological examination revealed that the sensitized treated PM groups had significant lower in inflammatory scores similar to dexamethasone treatments and the untreated group.. Administration of PM could inhibit airway inflammation, regulate cytokines, chemokines and enhance pulmonary conditions in allergic murine model of asthma.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Chemokines; Disease Models, Animal; Immunoglobulin E; Inflammation; Lung; Male; Methanol; Mice, Inbred BALB C; Mucus; Ovalbumin; Phytolaccaceae; Phytotherapy; Plant Extracts; Reactive Oxygen Species; Th2 Cells

We previously identified an immunomodulatory role of human induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells (MSCs) in asthmatic inflammation. Mitochondrial transfer from bone marrow MSCs to epithelial cells can result in the attenuation of acute lung injury in mice. However, the effects of mitochondrial transfer from iPSC-MSCs to epithelial cells in asthma and the mechanisms underlying these effects are unclear. We found that iPSC-MSC transplantation significantly reduced T helper 2 cytokines, attenuated the mitochondrial dysfunction of epithelial cells, and alleviated asthma inflammation in mice. Tunneling nanotubes (TNTs) were formed between iPSC-MSCs and epithelial cells, and mitochondrial transfer from iPSC-MSCs to epithelial cells via TNTs was observed both in vitro and in mice. Overexpression or silencing of connexin 43 (CX43) in iPSC-MSCs demonstrated that CX43 plays a critical role in the regulation of TNT formation by mediating mitochondrial transfer between iPSC-MSCs and epithelial cells. This study provides a therapeutic strategy for targeting asthma inflammation.

Topics: Animals; Apoptosis; Asthma; Cell Line; Cobalt; Connexin 43; Disease Models, Animal; Epithelial Cells; Humans; Induced Pluripotent Stem Cells; Inflammation; Lung; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Mice; Mitochondria; Nanotubes; Ovalbumin

Allergic asthma is a highly prevalent airway inflammatory disease, which involves the interaction between the immune system, environmental and genetic factors. Co-relation between allergic asthma and gut microbiota upon the change of diet have been widely reported, implicating that oral intake of alternative medicines possess a potential in the management of allergic asthma. Previous clinical, in vivo, and in vitro studies have shown that the Pentaherbs formula (PHF) comprising five traditional Chinese herbal medicines Lonicerae Flos, Menthae Herba, Phellodendri Cortex, Moutan Cortex, and Atractylodis Rhizoma possesses an anti-allergic and anti-inflammatory potential through suppressing various immune effector cells. In the present study, to further investigate the anti-inflammatory activities of PHF in allergic asthma, intragastrical administration of PHF was found to reduce airway hyperresponsiveness, airway wall remodeling and goblet cells hyperplasia in an ovalbumin (OVA)-induced allergic asthma mice model. PHF also significantly suppressed pulmonary eosinophilia and asthma-related cytokines IL-4 and IL-33 in bronchoalveolar lavage (BAL) fluid. In addition, PHF modulated the splenic regulatory T cells population, up-regulated regulatory interleukin (IL)-10 in serum, altered the microbial community structure and the short chain fatty acids content in the gut of the asthmatic mice. This study sheds light on the anti-inflammatory activities of PHF on allergic asthma. It also provides novel in vivo evidence that herbal medicines can ameliorate symptoms of allergic diseases may potentially prevent the development of subsequent atopic disorder such as allergic asthma through the influence of the gut microbiota.

Topics: Airway Remodeling; Animals; Anti-Inflammatory Agents; Asthma; Biodiversity; Cytokines; Disease Models, Animal; Drugs, Chinese Herbal; Eosinophils; Fatty Acids, Volatile; Gastrointestinal Microbiome; Immunoglobulin E; Male; Mice; Ovalbumin; Respiratory Hypersensitivity; Spleen; T-Lymphocytes, Regulatory

Asthma, a heterogeneous disease of the airways, is common around the world, but little is known about the molecular mechanisms underlying the interactions between DNA methylation and gene expression in relation to this disease. The seeds of

Topics: Animals; Anti-Asthmatic Agents; Asthma; Brassicaceae; Computational Biology; Disease Models, Animal; DNA Methylation; Epigenesis, Genetic; Gene Expression Profiling; Gene Regulatory Networks; Mice; Ovalbumin; Plant Extracts; Seeds; Transcriptome

The undesirable immune response poses a life-threatening challenge to human health. It not only deteriorates the therapeutic performance of biologic drugs but also contributes to various diseases such as allergies and autoimmune diseases. Inspired by the role of chromatin in the maintenance of natural immune tolerance, here we report a DNA-protein polymeric nanocomplex that can mimic the tolerogenic function of chromatin and induce an immune tolerance to its protein cargos. We first proved that the chromatin-mimetic nanomedicine loaded with keyhole limpet hemocyanin (KLH), a highly immunogenic model protein, could elicit a durable antigen-specific immune tolerance to KLH lasting for at least five weeks in mice. Following the proof-of-concept study, we demonstrated that this nanomedicine could be applied to improve the safety and efficacy of a biologic drug, PEGylated uricase, by attenuating the relevant antibody (Ab) responses. Moreover, we also demonstrated that prophylactic treatments with this nanomedicine could tolerize the immune system with the allergen of ovalbumin (OVA) and thus inhibit the occurrence of airway inflammation in an OVA-induced allergic asthma murine model. Collectively, our work illustrates a nature-inspired concept of immune tolerance induction and establishes a useful tool to specifically suppress unwanted immune responses for therapeutic purposes.

Topics: Animals; Asthma; Chromatin; Disease Models, Animal; Hemocyanins; Immune Tolerance; Male; Mice; Mice, Inbred C57BL; Nanomedicine; Ovalbumin

The etiology of asthma, which is a complicated disorder with various symptoms, including wheezing, coughing, and airflow obstruction, remains poorly understood. In addition, the effects of microRNAs (miRs) have not been explored. This study explored the effect of microRNA-200a (miR-200a) on airway smooth muscle cells (ASMCs) and airway remodeling in asthmatic mice. Furthermore, we speculated that miR-200a achieves its effect by targeting FOXC1 via the PI3K/AKT signaling pathway based on differentially expressed gene screening, target miR predictions and a bioinformatics analysis.. Eighty mice were assigned to a saline group or an ovalbumin (OVA) group, and the OVA group was transfected with a series of inhibitors, activators, and siRNAs to test the established mouse model. Airway reactivity and the ratio of eosinophils (EOSs) to leukocytes were detected. An ELISA was adopted to measure the levels of interleukin (IL)-4, IL-6, IL-8, tumor necrosis factor (TNF)-α, and immunoglobulin E (IgE). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting were used to determine the expression of FOXC1, PI3K, AKT, NF-κB, cyclin D1, TGF-β1 and p-AKT in ASMCs. Finally, CCK-8 assays were performed to detect cell proliferation and flow cytometry to detect apoptosis and cell cycle entry.. The bioinformatics analysis indicated that miR-200a mediated the PI3K/AKT signaling pathway by targeting FOXC1. In addition, mouse models of asthma were established. An elevated expression of miR-200a, a decreased mRNA and protein expression of FOXC1, PI3K, AKT, NF-κB, cyclin D1 and TGF-β1, a decreased expression of p-AKT, suppressed cell proliferation, accelerated apoptosis, and an increased number of cells at the G0/G1 phase were observed following the upregulation of miR-200a and downregulation of FOXC1.. The overexpression of miR-200a may downregulate FOXC1, thereby inhibiting the activation of the PI3K/AKT signaling pathway and ultimately suppressing ASMC proliferation and airway remodeling in asthmatic mice. This evidence supports the potential that miR-200a represents a new approach to treating asthma.

Topics: Airway Remodeling; Animals; Antagomirs; Asthma; Cell Proliferation; Disease Models, Animal; Female; Forkhead Transcription Factors; G1 Phase Cell Cycle Checkpoints; Mice; Mice, Inbred BALB C; MicroRNAs; Myocytes, Smooth Muscle; Ovalbumin; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; RNA Interference; RNA, Small Interfering; Signal Transduction; Tumor Necrosis Factor-alpha

Preclinical and clinical evidence indicates that a subset of asthma is driven by type 2 cytokines such as interleukin-4 (IL-4), IL-5, IL-9, and IL-13. Additional evidence predicts pathogenic roles for IL-6 and type I and type II interferons. Because each of these cytokines depends on Janus kinase 1 (JAK1) for signal transduction, and because many of the asthma-related effects of these cytokines manifest in the lung, we hypothesized that lung-restricted JAK1 inhibition may confer therapeutic benefit. To test this idea, we synthesized iJak-381, an inhalable small molecule specifically designed for local JAK1 inhibition in the lung. In pharmacodynamic models, iJak-381 suppressed signal transducer and activator of transcription 6 activation by IL-13. Furthermore, iJak-381 suppressed ovalbumin-induced lung inflammation in both murine and guinea pig asthma models and improved allergen-induced airway hyperresponsiveness in mice. In a model driven by human allergens, iJak-381 had a more potent suppressive effect on neutrophil-driven inflammation compared to systemic corticosteroid administration. The inhibitor iJak-381 reduced lung pathology, without affecting systemic Jak1 activity in rodents. Our data show that local inhibition of Jak1 in the lung can suppress lung inflammation without systemic Jak inhibition in rodents, suggesting that this strategy might be effective for treating asthma.

Topics: Administration, Inhalation; Allergens; Animals; Asthma; Dexamethasone; Disease Models, Animal; Eosinophils; Guinea Pigs; Inflammation; Janus Kinase 1; Lung; Ovalbumin; Protein Kinase Inhibitors; Signal Transduction; Treatment Outcome

Curcumin, a natural product found in the plant Curcuma longa, has been reported to have diverse range of molecular targets that influence numerous biochemical and molecular cascades including anti-inflammatory and antioxidant properties.. The aim of the study was to investigate the therapeutic potential of intranasal curcumin on ovalbumin (OVA)-induced chronic asthma and to elucidate underlying molecular mechanisms.. Mice were sensitized and exposed to 2% OVA aerosol for 2 times in a week for five consecutive weeks to study effect of intranasal curcumin on various MAPK pathway enzymes involved in chronic asthma and its effect on the activation of nuclear factor kB (NF-kB).. Curcumin treatment decreased the ROS level in BALF and nitrite level in blood serum of chronic asthmatic mice. Curcumin treatment had significantly decreased the phosphorylation of JNK, ERK1/2, and p38 and COX-2 expression thereby nuclear factor κB (NF-κB) activation and expression in lung tissues.. These results suggest that intranasal curcumin protects against asthma via action on mitogen-activated protein kinase (MAPK)/NF-κB signaling pathways.

Topics: Administration, Intranasal; Animals; Asthma; Curcuma; Curcumin; Cyclooxygenase 2; Lung; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinases; NF-kappa B; Ovalbumin; Phosphorylation; Signal Transduction

The activation of Toll-like receptor (TLR) signaling is widely reported to be involved in preventing the development of allergic asthma. However, the mechanism of the protective function of TLR signaling remains limited. Here, we studied the mouse model of ovalbumin (OVA)-induced allergic asthma and found that deficiency of TLR signaling or activating TLR signaling with agonist would aggravate or attenuate OVA-induced allergic asthma, respectively, and TLR signaling-mediated protective effect mainly affected the sensitization phase. After OVA/alum sensitization, neutrophils and inflammatory monocytes were recruited into peritoneal cavity and up-regulated TLRs expression. However, adoptive transfer of inflammatory monocytes but not peritoneal macrophages or neutrophils induced allergic symptoms in recipient mice after OVA challenge even without OVA/alum sensitization, and treating the inflammatory monocytes with TLR agonist

Topics: Allergens; Animals; Asthma; Cell Movement; Cells, Cultured; Cytokines; Disease Models, Animal; Humans; Hypersensitivity; Immunization; Inflammation Mediators; Mice; Mice, Inbred C57BL; Mice, Knockout; Monocytes; Neutrophils; Ovalbumin; Signal Transduction; Th1 Cells; Toll-Like Receptors

One of the most common comorbid pathology is asthma and arterial hypertension. For experimental modeling of comorbidity we have used spontaneously hypertensive rats with ovalbumin (OVA)-induced asthma. Rats were randomly divided into three groups: control group, OVA-induced asthma group; OVA-induced asthma + IL10 shRNA interference group. Target gene (IL10) was predicted by ANDSystem. We have demonstrated that RNA-interference of IL10 affected cardiovascular (tested using Millar microcatheter system) as well as respiratory functions (tested using force-oscillation technique, Flexivent) in rats. We have shown that during RNA-interference of IL10 gene in vivo there were changes in both cardiac and lung function parameters. These changes in the cardiovascular parameters can be described as positive. But the more intensive heart workload can lead to exhaust and decompensation of the heart functions. Knockdown of IL10 gene in asthma modeling induces some positive changes in respiratory functions of asthmatic animals such as decreased elastance and increased compliance of the lungs, as well as less pronounced pathomorphological changes in the lung tissue. Thus, we provide the data about experimentally confirmed functionality changes of the target which was in silico predicted to be associated with both asthma and hypertension - in our new experimental model of comorbid pathology.

Topics: Animals; Asthma; Comorbidity; Computational Biology; Hypertension; Interleukin-10; Male; Ovalbumin; Rats; Rats, Inbred SHR; RNA, Small Interfering

PI3K activation plays a central role in the development of pulmonary inflammation and tissue remodeling. PI3K inhibitors may thus offer an improved therapeutic opportunity to treat non-resolving lung inflammation but their action is limited by unwanted on-target systemic toxicity. Here we present CL27c, a prodrug pan-PI3K inhibitor designed for local therapy, and investigate whether inhaled CL27c is effective in asthma and pulmonary fibrosis. Mice inhaling CL27c show reduced insulin-evoked Akt phosphorylation in lungs, but no change in other tissues and no increase in blood glycaemia, in line with a local action. In murine models of acute or glucocorticoid-resistant neutrophilic asthma, inhaled CL27c reduces inflammation and improves lung function. Finally, inhaled CL27c administered in a therapeutic setting protects from bleomycin-induced lung fibrosis, ultimately leading to significantly improved survival. Therefore, local delivery of a pan-PI3K inhibitor prodrug reduces systemic on-target side effects but effectively treats asthma and irreversible pulmonary fibrosis.

Topics: Administration, Inhalation; Animals; Asthma; Benzene Derivatives; Bleomycin; Disease Models, Animal; Enzyme Inhibitors; Esters; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Proto-Oncogene Proteins c-akt; Pulmonary Fibrosis

Asthma is an inflammatory disease caused by an imbalance of Th1 and Th2 cells. In general, asthma is characterized by a stronger Th2 response. Most conventional asthma treatment focuses on improving airway flow or suppression of airway inflammation. To reduce the side effects of currently used asthma medicines, we have conducted studies on natural products that have no side effects. 2,3,5,4'-tetrahydroxystilbene-2-O-β-d-glucoside (TSG), the main compound of

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Disease Models, Animal; Female; Glucosides; Immunoglobulin Class Switching; Inflammation; Inflammation Mediators; Lung; Mice, Inbred C57BL; Ovalbumin; Protective Agents; Respiratory Hypersensitivity; Stilbenes; Th1 Cells; Th2 Cells

To study the effect of montelukast sodium (MK), a leukotriene receptor antagonist, and bacterial lysates (OM-85BV), used alone or in combination, on airway remodeling and the expression of transforming growth factor-β1 (TGF-β1) and Smad7 in guinea pigs with bronchial asthma and their correlation.. A total of 40 male Hartley guinea pigs were randomly divided into normal control group, asthma group, MK group, OM-85BV group, and MK+OM-85BV group, with 8 guinea pigs in each group. Intraperitoneal injection of 10% ovalbumin (OVA) for sensitization and aerosol inhalation of 1% OVA for challenge were performed to establish a model of airway remodeling of asthma in all of the groups apart from the normal control group, which were treated with normal saline. In the stage of challenge by aerosol inhalation, the guinea pigs in the MK, OM-85BV, and MK+OM-85BV groups were given corresponding suspension by gavage, and those in the normal control and asthma groups were given an equal volume of normal saline by gavage. Bronchoalveolar lavage fluid (BALF) of the guinea pigs was collected within 24 hours after challenge, and ELISA was used to measure the levels of TGF-β1 and Smad7 in BALF. The guinea pigs were sacrificed and the pathological section of lung tissue was prepared to observe the degree of airway remodeling. An image analysis technique was used to measure perimeter of the basement membrane (Pbm), total bronchial wall area (Wat), and airway bronchial smooth muscle area (Wam). Pearson linear regression was used to investigate the correlation between two variables.. According to the lung pathological section, compared with the normal control group, the asthma, MK, OM-85BV, and MK+OM-85BV groups had significant thickening of bronchial smooth muscle and alveolar wall, significantly higher Wat/Pbm and Wam/Pbm, a significantly higher level of TGF-β1, and a significantly lower level of Smad7 (P<0.05). Compared with the asthma group, the MK, OM-85BV, and MK+OM-85BV groups had a significant improvement in pathological injury, significantly lower Wat/Pbm and Wam/Pbm, a significantly lower level of TGF-β1, and a significantly higher level of Smad7 (P<0.05). The MK+OM-85BV group had significantly greater improvements than the MK group and the OM-85BV group (P<0.05). The expression of TGF-β1 was negatively correlated with that of Smad7 and positively correlated with Wat/Pbm and Wam/Pbm, and the expression of Smad7 was negatively correlated with Wat/Pbm and Wam/Pbm (P<0.01).. MK and OM-85BV, used alone or in combination, can reduce airway remodeling in guinea pigs with asthma, and MK combined with OM-85BV has the best effect, possibly by reducing TGF-β1 expression, increasing Smad7 expression, and improving the TGF-β1/Smad7 imbalance.

Topics: Acetates; Airway Remodeling; Animals; Asthma; Cell Extracts; Cyclopropanes; Guinea Pigs; Lung; Male; Ovalbumin; Quinolines; Sulfides; Transforming Growth Factor beta1

Asthma is a chronic inflammatory disease of airways mediated by T-helper 2 (Th2) cells involving complex signaling pathways. Although resveratrol has previously been shown to attenuate allergic asthma, the role of miRNA in this process has not been studied. We investigated the effect of resveratrol on ovalbumin-induced experimental allergic asthma in mice. To that end, BALB/c mice were immunized with ovalbumin (OVA) intraperitoneally followed by oral gavage of vehicle (OVA-veh) or resveratrol (100 mg/kg body) (OVA-res). On day 7, the experimental groups received intranasal challenge of OVA followed by 7 days of additional oral gavage of vehicle or resveratrol. At day 15, all mice were euthanized and bronchioalveolar fluid (BALF), serum and lung infiltrating cells were collected and analyzed. The data showed that resveratrol significantly reduced IL-5, IL-13, and TGF-β in the serum and BALF in mice with OVA-induced asthma. Also, we saw a decrease in CD3+CD4+, CD3+CD8+, and CD4+IL-4+ cells with increase in CD4+CD25+FOXP3+ cells in pulmonary inflammatory cell infiltrate in OVA-res group when compared to OVA-veh. miRNA expression arrays using lung infiltrating cells showed that resveratrol caused significant alterations in miRNA expression, specifically downregulating the expression of miR-34a. Additionally, miR-34a was found to target

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Down-Regulation; Female; Forkhead Transcription Factors; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; MicroRNAs; Ovalbumin; Resveratrol; T-Lymphocytes, Regulatory; Treatment Outcome

Asthma is a common chronic inflammatory disease in which lung airways narrow and produce extra mucus. Numerous medications, such as steroids, are used to prevent or control asthmatic symptoms, but side effects are associated with those medications. There are reports of anti-inflammatory, antibacterial, and antiparasitic effects of terpene, a volatile organic compound (VOC) in conifers. VOCs easily enter a gaseous form, and wood products are good sources of VOCs. However, only a few studies have been conducted on the effect on asthma of VOCs emitted by wood. In this study, we examined the effects of VOCs diffused from wood panels on ovoalbumin (OVA)-induced asthma in a mouse model. The mice were intraperitoneally sensitized with 10 μg of OVA with aluminum hydroxide on days 0, 7, and 14. From day 21 to day 26, the mice were challenged with 2% OVA intranasally for 30 min. For VOC treatment, asthma model mice were placed in polyacrylamide chambers containing wood panels of Chamaecyparis obtusa, Pinus densiflora, Pinus koraiensis, or Larix kaempferi. On day 27, serum, lung tissue, and bronchoalveolar lavage fluids were prepared for H&E staining, qRT-PCR, ELISA, and Diff-Quik staining, as appropriate. OVA treatment induced hypertrophy of the bronchiolar wall. The budesonide group and all four of the wood panel-exposed groups showed less thickening of the bronchiolar wall and downregulated transcriptional expressions of cytokines such as interleukin-4 (IL-4) and interleukin-13 (IL-13). The serum tumor necrosis factor-α (TNF-α) mRNA expression level was significantly decreased only in the C. obtusa group, but the serum IL-4 levels were decreased in all wood panel treatment groups. Diff-Quik staining of bronchoalveolar lavage fluids revealed a decrease in the number of granulocytes in all wood panel treatment groups. The results suggest that VOCs from C. obtusa, P. densiflora, P. koraiensis and L. kaempferi produce antiasthmatic effects by regulating the production of IL-4, IL-9, IL-13, TNF-α.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Budesonide; Chamaecyparis; Cytokines; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Larix; Mice; Mice, Inbred ICR; Ovalbumin; Pinus; Reverse Transcriptase Polymerase Chain Reaction; Volatile Organic Compounds; Wood

It has been suggested that asthmatics are more susceptible than healthy individuals to airborne irritating chemicals in general. However, there is limited human data available to support this hypothesis due to ethical and practical difficulties. We explored a murine model of ovalbumin (OVA)-induced airway inflammation to study susceptibility during acute exposure to chemicals with chlorine as a model substance.. Naïve and OVA sensitized female BALB/c mice were exposed to chlorine at four different concentrations (0, 5, 30 and 80 ppm) for 15 minutes with online recording of the respiratory function by plethysmography. The specific effects on respiratory mechanics, inflammatory cells and inflammatory mediators (cytokines and chemokines) of the airways were measured 24 hours after the chlorine exposure as well as histopathological examination of the lungs.. Similar concentration-dependent reductions in respiratory frequency were seen in the two groups, with a 50% reduction (RD. The results do not support an increased susceptibility to chlorine among OVA-sensitized mice. This animal model, which represents a phenotype of eosinophilic airway inflammation, seems unsuitable to study susceptibility to inhalation of irritants in relation to asthma.

Topics: Administration, Inhalation; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Chlorine; Cytokines; Disease Models, Animal; Female; Immunoglobulin E; Irritants; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Mechanics

There has been an increase in the popularity of waterpipe tobacco smoking (WTS) worldwide, especially in the younger population, including asthma patients. In this study, we investigated the effects of waterpipe smoking on airway inflammation, cytokine levels and oxidative stress markers in an antigen-driven murine model of asthma.. Balb/c mice were divided into four groups; (1) control (received fresh air, ovalbumin sensitization and saline challenge), (2) WTS (received WTS, ovalbumin sensitization and saline challenge), (3) Ova S/C (received fresh air, ovalbumin sensitization and ovalbumin challenge) and (4) simultaneous WTS and Ova S/C (received WTS, ovalbumin sensitization and ovalbumin challenge). Airway inflammatory cells were evaluated in the broncho-alveolar lavage fluid. Cytokines [interleukin (IL)-13, 10 and 18] and oxidative stress markers [superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx)] were evaluated in the lung homogenates.. Chronic exposure to WTS significantly increased the number of airway inflammatory cells in mice, specifically: eosinophils, neutrophils, macrophages and lymphocytes. The level of IL-13 in the lungs was increased and the level of IL-10 was reduced (p < 0.05) by WTS. Chronic WTS potentiated the increase in inflammatory cells induced by Ova S/C (p < 0.05). The level of IL-13 in the lungs was increased by simultaneous WTS and Ova S/C (p < 0.05) while, levels of IL-10, IL-18, SOD, catalase and GPx in the lungs were not affected.. Chronic WTS exposure induced airway inflammation in control mice and enhanced airway inflammation in murine model of asthma.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Catalase; Cytokines; Disease Models, Animal; Glutathione Peroxidase; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Superoxide Dismutase; Water Pipe Smoking

Echinodorus scaber, Alismataceae, is popularly known in Brazil as "chapéu-de-couro". The plant leaves are used by the population as decoction, infusion, or maceration in bottled spirits, to treat inflammatory respiratory diseases.. To investigate the anti-inflammatory mechanism of the hydroethanolic extract of leaves of Echinodorus scaber (HEEs) in allergic asthma. A phytochemical analysis of the extract was performed as well.. HEEs reduced total leukocyte, eosinophil, neutrophil, and mononuclear cell counts at all doses tested, with maximum effect at 30mg/kg (73.9%, 75.9%, 75.5%, and 65.2% reduction, p<0.001, respectively). Increases in T. Our findings provided pharmacological preclinical evidence for the popular use of the leaves of Echinodorus scaber in allergic inflammation. Its anti-inflammatory effect was dependent on the decrease in migratory inflammatory cells, and both T

Topics: Alismataceae; Animals; Anti-Inflammatory Agents; Asthma; Brazil; Bronchoalveolar Lavage Fluid; Chromatography, High Pressure Liquid; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Inflammation; Male; Mice; Ovalbumin; Plant Extracts; Plant Leaves

ORMDL sphingolipid biosynthesis regulator 3 (ORMDL3) has been previously implicated in asthma pathogenesis, its effect on airway remodeling remains to be elucidated. The present study examined the expression levels of ORMDL3 in a mouse model of asthma. Mice were divided into three groups: Asthmatic model (n=10), budesonide‑treated (n=10) and a control group (n=8). Asthma was induced by sensitization with ovalbumin (OVA) and aluminum hydroxide on day 1, 7 and 14. Subsequently mice were exposed to OVA three times per week from day 28. In order to investigate the mechanism of airway remodeling 100 µg/kg aerosol budesonide was administered to 6 animals prior to exposure to OVA. The condition of lung tissues was assessed through histology, and the expression levels of ORMDL3, phosphorylated‑extracellular‑signal regulated kinase (p‑ERK) and matrix metallopeptidase‑9 (MMP‑9) were quantified using immunohistochemistry, reverse transcription‑quantitative polymerase chain reaction and western blotting. A severe inflammatory response and airway remodeling were pretreatment with budesonide. Expression levels of ORMDL3, phosphorylated (p)‑ERK and MMP‑9 were significantly greater in the asthma‑model group; however, in the group pretreated with budesonide their expression was reduced. Expression levels of ORMDL3, p‑ERK and MMP‑9 were significantly positively correlated with bronchial wall thickness. ORMDL3 expression was significantly positively correlated with p‑ERK and MMP‑9. Therefore, increased ORMDL3 expression may induce the p‑ERK/MMP‑9 pathway to promote pathological airway remodeling in patients with asthma.

Topics: Airway Remodeling; Aluminum Hydroxide; Animals; Asthma; Budesonide; Collagen; Disease Models, Animal; Extracellular Signal-Regulated MAP Kinases; Female; Immunohistochemistry; Lung; Matrix Metalloproteinase 9; Membrane Proteins; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphorylation; Real-Time Polymerase Chain Reaction; Signal Transduction

To find bioactive medicinal herbs exerting anti-asthmatic activity, we investigated the effect of an aqueous extract of Urtica dioica L. (Urticaceae) leaves (UD), the closest extract to the Algerian traditional use.. In this study, we investigated the in vivo anti-asthmatic and antioxidant activities of nettle extract.. Adult male Wistar rats were divided into four groups: Group I: negative control; group II: Ovalbumin sensitized/challenged rats (positive control); group III: received UD extract (1.5 g/kg/day) orally along the experimental protocol; group IV: received UD extract (1.5 g/kg/day) orally along the experimental protocol and sensitized/challenged with ovalbumin. After 25 days, blood and tissue samples were collected for haematological and histopathological analysis, respectively. The oxidative stress parameters were evaluated in the lungs, liver and erythrocytes. Then, correlations between markers of airway inflammation and markers of oxidative stress were explored.. UD extract significantly (p < 0.01) inhibited eosinophilia increases in BALF (-60%) and the levels of leucocytes (-32.75%) and lymphocytes (-29.22%) in serum, and effectively suppressed inflammatory cells recruitment in the asthmatic rat model. Besides, the lipid peroxidation generated by allergen administration was significantly (p < 0.05) diminished by UD treatment in lung tissue (-48.58%). The nettle extract was also investigated for the total phenolic content (30.79 ± 0.96 mg gallic acid/g dry extract) and shows DPPH radical scavenging activity with 152.34 ± 0.37 μg/mL IC. The results confirmed that UD administration might be responsible for the protective effects of this extract against airway inflammation.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Antioxidants; Asthma; Biphenyl Compounds; Disease Models, Animal; Lipid Peroxidation; Lung; Male; Ovalbumin; Oxidative Stress; Phenols; Phytotherapy; Picrates; Plant Extracts; Plant Leaves; Plants, Medicinal; Pneumonia; Pulmonary Eosinophilia; Rats, Wistar; Urtica dioica

Aerobic exercise (AE) reduces lung function decline and risk of exacerbations in asthmatic patients. However, the inflammatory lung response involved in exercise during the sensitization remains unclear. Therefore, we evaluated the effects of exercise for 2 weeks in an experimental model of sensitization and single ovalbumin-challenge. Mice were divided into 4 groups: mice non-sensitized and not submitted to exercise (Sedentary, n=10); mice non-sensitized and submitted to exercise (Exercise, n=10); mice sensitized and exposed to ovalbumin (OVA, n=10); and mice sensitized, submitted to exercise and exposed to OVA (OVA+Exercise, n=10). 24 h after the OVA/saline exposure, we counted inflammatory cells from bronchoalveolar fluid (BALF), lung levels of total IgE, IL-4, IL-5, IL-10 and IL-1ra, measurements of OVA-specific IgG1 and IgE, and VEGF and NOS-2 expression via western blotting. AE reduced cell counts from BALF in the OVA group (p<0.05), total IgE, IL-4 and IL-5 lung levels and OVA-specific IgE and IgG1 titers (p<0.05). There was an increase of NOS-2 expression, IL-10 and IL-1ra lung levels in the OVA groups (p<0.05). Our results showed that AE attenuated the acute lung inflammation, suggesting immunomodulatory properties on the sensitization process in the early phases of antigen presentation in asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Immunoglobulin E; Immunoglobulin G; Interleukins; Male; Mice; Nitric Oxide Synthase Type II; Ovalbumin; Physical Conditioning, Animal; Pneumonia; Vascular Endothelial Growth Factor A

Ovalbumin-induced allergic lung inflammation (ALI) is a condition believed to be mediated by cytokines, extracellular matrix remodeling, and redox imbalance. In this study, we evaluated pulmonary function together with inflammatory markers as interleukin-4 (IL-4), myeloperoxidase (MPO), eosinophil cells, and redox markers in the lungs of BALB/c mice after ovalbumin (OVA) sensitization and challenge. Our results showed an increase in bronchial hyperresponsiveness stimulated by methacholine (Mch), inflammatory cell influx, especially eosinophils together with an increase of high mobility group box 1 (HMGB1) and altered lipid peroxidation (LP) and antioxidant defenses in the OVA group compared to the control group (p ≤ 0.5). Thus, we demonstrated that OVA-induced ALI altered redox status concomitantly with impaired lung function, which was associated with HMGB1 expression and proteolytic remodeling. Taken together all results found here, we may suggest HMGB1 is an important therapeutic target for asthma, once orchestrates the redox signaling, inflammation, and remodeling that contribute to the disease development.

Topics: Animals; Asthma; Biomarkers; Bronchial Hyperreactivity; Eosinophils; HMGB1 Protein; Inflammation; Lipid Peroxidation; Mice; Mice, Inbred BALB C; Ovalbumin; Oxidative Stress; Proteolysis

Dihydromyricetin (DHM) is a plant flavonoid and is the primary active ingredient isolated from the medicinal herb, Ampelopsis grossedentata. DHM has been shown to possess various pharmacological activities, including anti‑inflammatory effects. However, the possible role of DHM in asthma treatment remains to be elucidated. The present study aimed to investigate its anti‑inflammatory properties in mice with symptoms of allergic asthma. The C57BL/6 mice were sensitized and challenged with ovalbumin (OVA) to induce asthma. DHM or phosphate‑buffered saline treatment was administered 1 h prior to the OVA challenge. The levels of interleukin (IL)‑4, IL‑5 and IL‑13 in the bronchoalveolar lavage (BAL) fluid were measured by enzyme‑linked immunosorbent assay (ELISA), and OVA‑specific serum IgE and IgG1 levels were also determined by ELISA. Histopathological staining was performed to evaluate the infiltration of inflammatory cells into the BAL fluid, lung tissues and goblet cell hyperplasia. DHM treatment significantly reduced the total number of inflammatory cells, including eosinophils, neutrophils, lymphocytes and macrophages, in the BAL fluid. DHM also reduced the levels of IL‑4, IL‑5 and IL‑13 in the BAL fluid, and reduced the secretion of OVA‑specific IgE and IgG1 in the serum. The histological staining demonstrated that DHM treatment effectively suppressed the OVA‑induced inflammatory cells in the lung tissues and in the mucus hypersecreted by goblet cells in the airway. These results showed that DHM had a potent anti‑inflammatory effect in an OVA‑induced mouse model of asthma, offering potential as an anti‑inflammatory agent for the treatment of asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Flavonols; Goblet Cells; Immunoglobulin E; Immunoglobulin G; Mice; Mucus; Ovalbumin

CCL11, a chemokine, is linked to the early development of airways eosinophilia in allergic asthma. Therefore, CCL11 production is a target for abrogating eosinophilic-driven airway inflammation. Blackcurrants are high in compounds that regulate inflammation, particularly anthocyanins. In this study, we investigated the effect of oral blackcurrant supplementation on allergen-induced eosinophilia and CCL11 production; we also profiled key compounds in blackcurrants that were linked to this effect. Ten milligram per kilogram (total anthocyanins) of a commercially available, anthocyanin-rich New Zealand "Ben Ard" blackcurrant extract ("Currantex 30") attenuated ovalbumin-induced inflammation, eosinophilia (by 52.45 ± 38.50%), and CCL11 production (by 48.55 ± 28.56%) in a mouse model of acute allergic lung inflammation. Ten blackcurrant polyphenolic extracts were also found to suppress CCL11 secretion by stimulated human lung epithelial cells in vitro. Correlation analysis identified potential blackcurrant polyphenolic anthocyanin constituents specifically delphinidins and cyanidins, involved in CCL11 suppression. Our findings show oral supplementation with New Zealand blackcurrant is effective in reducing lung inflammation, and highlight the potential benefit of developing cultivars with specific polyphenolic profiles for the creation of functional foods with desirable biological activity.

Topics: Animals; Anthocyanins; Asthma; Cells, Cultured; Chemokine CCL11; Male; Mice; Mice, Inbred C57BL; Ovalbumin; Plant Extracts; Ribes

Objective To investigate the effect of estrogen on the latent period of inducing asthma and airway inflammation in mice with asthma. Methods Forty male BALB/c mice were randomly divided into normal control group, OVA-induced asthma group, 400 μg/kg estradiol treatment group, 400 μg/kg estradiol combined with 7 mg/kg tamoxifen group, with 10 mice in each group. The OVA-induced asthma group, estradiol treatment group, and estradiol combined with tamoxifen group were sensitized and challenged by OVA; the normal control group was treated with normal saline instead. The estradiol treatment group and estradiol combined with tamoxifen group were given intraperitoneal injection of 400 μg/kg estradiol 4 hours before each challenge, and the latter was given additionally intraperitoneal injection of 7 mg/kg tamoxifen at 30 minutes before the injection of estradiol; the normal control group and OVA-induced asthma group were injected with normal saline as controls. The latent period of inducing asthma of all the mice was determined 24 hours after the final OVA challenge. Bronchoalveolar lavage fluid (BALF) was collected for counting total leukocyte cells and the percentages of eosinophils and lymphocytes. The concentrations of interleukin 4 (IL-4) and IL-13 in BALF were detected by ELISA. The pathologic change of lung tissues was examined by HE staining. Results Compared with the normal control group, the latent period of inducing asthma were shortened in the OVA-induced asthma group, estradiol treatment group, and estradiol combined with tamoxifen group; compared with the OVA-induced asthma group and estradiol combined with tamoxifen group, the latent period of inducing asthma of the estradiol treatment group were shortened; there was no significant difference in the latent period between the OVA-induced asthma group and the estradiol combined with tamoxifen group. Compared with the normal control group, the levels of IL-4 and IL-13 increased in the OVA-induced asthma group, estradiol treatment group, and estradiol combined with tamoxifen group; compared with the OVA-induced asthma group and estradiol combined with tamoxifen group, the levels of IL-4 and IL-13 increased in the estradiol treatment group; there was also no significant difference between the OVA-induced asthma group and the estradiol combined with tamoxifen group. Compared with the normal control group, the total leukocytes and the percentages of eosinophils and lymphocytes in BALF were raised in the OV

Topics: Animals; Asthma; Bronchi; Disease Models, Animal; Eosinophils; Estrogens; Female; Humans; Interleukin-13; Interleukin-4; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin

Chrysophanol (CH), extracted from plants of Rheum genus, possesses various pharmacological effects including anti-inflammatory activity. The purpose of the present study was to evaluate the protective effects and the underlying mechanisms of CH on ovalbumin (OVA)-induced asthma in mice. Fifty mice were randomly assigned to five experimental groups: control group, model group, dexamethasone (2 mg/kg) group and CH (5 and 10 mg/kg) groups. The number of eosinophil cells and the production of interleukin-6 (IL-6), IL-1β, IL-17 A and tumor necrosis factor-α in bronchoalveolar lavage fluid (BALF) were measured. In addition, pulmonary histopathology, airway resistance (Raw), T-helper17 (Th17) cells frequency and RORγt expression were evaluated. Our study demonstrated that CH effectively decreased eosinophil count and inflammatory cytokines production in BALF. In addition, treatment with CH significantly inhibited the Raw, Th17 percentage and RORγt expression in OVA-induced animals compared with those in model group. Histological studies also demonstrated that CH significantly suppressed OVA-induced eosinophilia in lung tissue compared with model group. Our findings supported that CH can prevent allergic asthma in the mouse model.

Topics: Airway Resistance; Animals; Anthraquinones; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Flow Cytometry; Mice, Inbred BALB C; Ovalbumin; Rheum; Th17 Cells

To determine whether oral administration of. Ovalbumin (OVA)-induced allergic asthma and β-lactoglobulin-induced food allergy mouse models were used in this study. Following oral administration of. Oral administration of

Topics: Allergens; Animals; Asthma; Bifidobacterium longum subspecies infantis; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Food Hypersensitivity; Immunoglobulin E; Immunoglobulin G; Inflammation; Interleukin-13; Interleukin-4; Intestines; Lactoglobulins; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics

18β-Glycyrrhetinic acid (18Gly), the major bioactive component of Glycyrrhizae Radix, possesses anti-ulcerative, anti-inflammatory, and other pharmacological properties. Although 18Gly is associated with immunoregulatory functions of allergic diseases, the pathophysiological mechanisms of 18Gly action in allergic inflammatory lung disease have not been examined. Moreover, there are no in vivo studies on the anti-asthmatic effects of 18Gly in allergic asthma. We investigated its effect and mechanism of action in airway inflammation in a BALB/c mouse model of allergic asthma. Interestingly, 18Gly strongly suppressed airway hyperresponsiveness, accumulation of inflammatory cells, and levels of T helper type 2 (Th2) cytokines (interleukin (IL)-5 and IL-13) in bronchoalveolar lavage fluid (BALF). It also attenuated lung IL-5, IL-13, and IL-4 expression, but it upregulated peroxisome proliferator-activated receptor gamma (PPARγ) mRNA expression in lungs. Moreover, it exerted immunomodulatory effects by suppressing Th2 cytokines (IL-5, IL-13) production through upregulation of forkhead box p3 (Foxp3), and downregulation of signal transducer and activator of transcription (STAT6), GATA-binding protein 3 (GATA-3), and retinoic acid-related orphan receptor γ t (RORγt) expression. These results suggest that the anti-asthmatic activity of 18Gly may occur by the suppression of IL-5, IL-13, and OVA-specific Immunoglobulin E (IgE) production through inhibition of the RORγt, STAT6, GATA-3 pathways and upregulation of the Foxp3 transcription pathway. Also, 18Gly treatment was protective against the oxidative stress by inducing significant decrease of reactive oxygen species (ROS) generation in MH-S alveolar macrophage cells. Our results suggest that 18Gly can improve allergic asthma and can be a novel therapeutic component for the treatment of allergic asthma.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cell Line; Cell Survival; Cytokines; Fabaceae; Female; Forkhead Transcription Factors; GATA3 Transcription Factor; Glycyrrhetinic Acid; Immunoglobulin E; Lung; Mice, Inbred BALB C; Ovalbumin; Phytotherapy; Reactive Oxygen Species; Spleen; STAT6 Transcription Factor; Th2 Cells

Adenosine induces bronchial hyperresponsiveness and inflammation in asthmatics through activation of A

Topics: Adenosine A2 Receptor Antagonists; Animals; Asthma; Dogs; Drug Design; Hep G2 Cells; Humans; Madin Darby Canine Kidney Cells; Mice; Mice, Inbred C57BL; Microsomes, Liver; Molecular Docking Simulation; Ovalbumin; Rats; Receptor, Adenosine A2B; Xanthine

Since asthma is a multifactorial disease where treatment sometimes is not effective, new therapies that improve the respiratory discomfort of patients are of great importance. Phototherapy as Light-emitting diode (LED) has emerged as a treatment that presents good results for diseases that are characterized by inflammation. Thus, our objective was to investigate the effects of LED on lung inflammation, by an evaluation of lung cell infiltration, mucus secretion, oedema, and the production of cytokines. Male Balb/c mice were or not sensitized and challenged with ovalbumin (OVA) and treated or not with LED therapy (1 h and 4 h after each OVA challenge). Twenty-four hours after the last OVA challenge, analyzes were performed. Our results showed that LED treatment in asthmatic mice reduced the lung cell infiltration, the mucus production, the oedema, and the tracheal's contractile response. It also increased the IL-10 and the IFN-gamma levels. The effects of LED treatment on lung inflammation may be modulated by IL-10, IFN-gamma, and by mast cells. This study may provide important information about the effects of LED, and in addition, it may open the possibility of a new approach for the treatment of asthma.

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Count; Cytokines; Disease Models, Animal; Granulocytes; Lymphocytes; Macrophages; Male; Mast Cells; Mice; Mice, Inbred BALB C; Muscle Contraction; Ovalbumin; Phototherapy; Pneumonia; Trachea

We describe pharmacokinetic and pharmacodynamic properties of two novel oral drug candidates for asthma. Phenolic α

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Eosinophils; Flow Cytometry; Humans; Lung; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, GABA; Respiratory Hypersensitivity; Swine

Bacillus Calmette-Guerin (BCG) is a potent agent for the prevention of tuberculosis. Current studies have regarded BCG as an immunomodulator. However, there is little information on whether it can be used to inhibit airway inflammation and airway remodeling caused by asthma. Therefore, in this study, we investigate the role of epithelial-mesenchymal transition (EMT) in airway inflammation and airway remodeling as well as the possible therapeutic mechanism of BCG for the treatment of asthma. Wistar rats were sensitized and challenged by ovalbumin for 2 weeks or 8 weeks. BCG was subcutaneously administered daily before every ovalbumin challenge to determine its therapeutic effects. The 2 weeks model group showed extensive eosinophilia, chronic inflammatory responses, bronchial wall thickening, airway epithelium damage, increased levels of transforming growth factor β 1 (TGF-β

Topics: Actins; Airway Remodeling; Animals; Asthma; BCG Vaccine; Bronchi; Bronchoalveolar Lavage Fluid; Cadherins; Disease Models, Animal; Epithelial-Mesenchymal Transition; Fibronectins; Goblet Cells; Humans; Inflammation; Ovalbumin; Rats; Rats, Wistar; Transforming Growth Factor beta; Transforming Growth Factor beta1

Topics: Animals; Anti-Allergic Agents; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bone Marrow Cells; Cell Degranulation; Dermatitis, Atopic; Disease Models, Animal; Female; Immunoglobulin E; Lactones; Leukotriene C4; Mast Cells; Mice; Mice, Inbred BALB C; Ovalbumin; Phosphorylation; Respiratory Hypersensitivity; Sesquiterpenes; Signal Transduction

Present study was performed to examine whether ADH was implicated in psychological stress asthma and to explore the underlying molecular mechanism.. We not only examined ADH levels in the cerebrospinal fluid (CSF) via radioimmunoassay, but also measured ADH receptor (ADHR) expression in airway-related vagal preganglionic neurons (AVPNs) through real-time PCR in all experimental mice. Western blotting was performed to evaluate the relationship between ADH and PKA/PKC in psychological stress asthma. Finally, the role of PKA/PKC in psychological stress asthma was analyzed.. Marked asthma exacerbations were noted owing to significantly elevated levels of ADH and ADHR after psychological stress induction as compared to OVA alone (asthma group). ADHR antagonists (SR-49095 or SR-121463A) dramatically lowered higher protein levels of PKAα and PKCα induced by psychological stress as compared to OVA alone, suggesting the correlation between ADH and PKA/PKC in psychological stress asthma. KT-5720 (PKA inhibitor) and Go-7874 (PKC inhibitor) further directly revealed the involvement of PKA/PKC in psychological stress asthma. Some notable changes were also noted after employing PKA and PKC inhibitors in psychological stress asthma, including reduced asthmatic inflammation (lower eosinophil peroxidase (EPO) activity, myeloperoxidase (MPO) activity, immunoglobulin E (IgE) level, and histamine release), substantial decrements in inflammatory cell counts (eosinophils and lymphocytes), and decreased cytokine secretion (IL-6, IL-10, and IFN-γ), indicating the involvement of PKA/PKC in asthma exacerbations induced by psychological stress.. Our results strongly suggested that ADH participated in psychological stress-induced asthma exacerbations via PKA/PKC signal pathway in AVPNs.

Topics: Animals; Antidiuretic Hormone Receptor Antagonists; Asthma; Carbazoles; Cyclic AMP-Dependent Protein Kinases; Cytokines; Disease Models, Animal; Eosinophils; Female; Mice; Mice, Inbred BALB C; Morpholines; Neurons; Ovalbumin; Protein Kinase C; Protein Kinase Inhibitors; Pyrroles; Receptors, Vasopressin; Signal Transduction; Spiro Compounds; Stress, Psychological; Vasopressins

This study explored the effects of microRNA-19b (miR-19b) on airway remodeling, airway inflammation, and degree of oxidative stress in a mouse model of asthma. Bioinformatics analyses and dual luciferase reporter gene assays revealed that thymic stromal lymphopoietin (TSLP) is a direct target of miR-19b. An asthma model was established via ovalbumin (OVA) sensitization and challenge in 72 female BALB/c mice. Mice were then assigned to saline, OVA-sensitized, saline+miR-19b mimics, saline+anti-TSLP, OVA-sensitized+miR-19b mimics, OVA-sensitized+mimics scramble, OVA-sensitized+anti-TSLP, and OVA-sensitized+IgG2a groups. Pathological morphology changes were detected through hematoxylin/eosin, Masson, and periodic acid-Schiff staining. miR-19b was downregulated while TSLP and Stat3 were upregulated in the OVA-sensitized group compared with the saline group. Bronchoalveolar lavage fluid samples from OVA-sensitized mice showed increased total protein, IL-4, IL-5 and IL-6 levels, numbers of inflammatory cells, eosinophils, neutrophils, mononuclear macrophages and lymphocytes, and eosinophil% compared to controls. Lung tissues from sensitized mice exhibited decreased superoxide dismutase activity and increased methane dicarboxylic aldehyde levels. The effects of OVA sensitization were reversed in the OVA-sensitized+miR-19b mimics and OVA-sensitized+anti-TSLP groups. These findings suggest miR-19b reduces airway remodeling, airway inflammation, and degree of oxidative stress by inhibiting Stat3 signaling through TSLP downregulation in a mouse asthma model.

Topics: 3' Untranslated Regions; Airway Remodeling; Animals; Asthma; Biomarkers; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Female; Gene Expression Regulation; Inflammation Mediators; Mice; MicroRNAs; Ovalbumin; Oxidation-Reduction; Oxidative Stress; RNA Interference; Signal Transduction; STAT3 Transcription Factor; Thymic Stromal Lymphopoietin

Asthma is a chronic disease related to airway hyperresponsiveness and airway remodeling. Airway remodeling is the important reason of refractory asthma and is associated with differentiation of airway epithelia into myofibroblasts via epithelial-mesenchymal transition (EMT) to increase the process of subepithelial fibrosis. There is growing evidence that autophagy modulates remodeling. However, the underlying molecular mechanisms of these effects are still unclear. In this study, we hypothesized that Follistatin-like 1 (FSTL1) promotes EMT and airway remodeling by intensifying autophagy. With the use of transmission electron microscopy (TEM), double-membrane autophagosomes were detected in the airways of patients and mice. More autophagosomes were in patients with asthma and OVA-challenged mice compared with healthy controls. The expression of FSTL1 and beclin-1 was upregulated in the airways of patients with asthma and OVA-challenged mice, accompanied by airway EMT and remodeling. In OVA-challenged

Topics: Adult; Airway Remodeling; Animals; Asthma; Autophagosomes; Autophagy; Biomarkers; Bronchoalveolar Lavage Fluid; Chromones; Disease Models, Animal; Epithelial-Mesenchymal Transition; Female; Follistatin-Related Proteins; Humans; Lung; Male; Mice, Inbred C57BL; Morpholines; Ovalbumin; Rats; Respiratory Hypersensitivity; Up-Regulation

Topics: Adoptive Transfer; Animals; Asthma; Basophils; Cell Differentiation; Cells, Cultured; Coculture Techniques; Disease Models, Animal; Female; Genotype; Immunoglobulin J Recombination Signal Sequence-Binding Protein; Lung; Lymphocyte Activation; Membrane Glycoproteins; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; OX40 Ligand; Phenotype; Receptors, Notch; RNA, Messenger; Signal Transduction; Th1 Cells; Th2 Cells; Tumor Necrosis Factors

Recently the role of gastrointestinal nematodes in modulating the immune responses in inflammatory and immune-mediated conditions such as allergy and autoimmune diseases has been introduced. This is mainly due to the suppressive effects of somatic and excretory secretory (ES) products of nematodes on the immune responses. In this study, we evaluated the immunomodulatory potentials of somatic products of Marshallagia marshalli, a gastrointestinal nematodes of sheep, to suppress the immune-mediated responses in a murine model of allergic airway inflammation. BALB/c mice were intraperitoneally (IP) sensitized with ovalbumin (OVA)/Alum and then challenged with 1% OVA. Somatic products of M. marshalli were administered during each sensitization. The effects of somatic products on development of allergic airway inflammation were evaluated by analyzing inflammatory cells recruitment, histopathological changes, cytokines production (IL-4, IL-13, IL-10, TGF-β) and serum antibody titers (IgG1, IgG2a).. Somatic products of M. marshalli were able to suppress the induction of allergic airway inflammation in mice. Modulation of Th2 type responses (IL-4, IL-13, IgG1) via upregulations of IL-10 and TGF-β production was observed after injection of somatic products of M. marshalli. In addition, inflammatory cells infiltration and pathological disorders were significantly diminished following administration of somatic products.. Our data raised the possibility that helminths could be a potential therapeutic candidate to alleviate the inflammatory conditions in allergic asthma. According to these results, we concluded that M. marshalli may contain immune-modulatory molecules that attenuate allergic airway inflammation via induction of regulatory cytokines. Further investigations are required to identify molecules that might have potentials for development of novel therapeutic targets.

Topics: Alum Compounds; Animals; Asthma; Cytokines; Disease Models, Animal; Down-Regulation; Hypersensitivity; Immunization; Immunomodulation; Interleukin-10; Interleukin-4; Mice; Mice, Inbred BALB C; Ovalbumin; Sheep; Th2 Cells; Tissue Extracts; Trichostrongyloidea

Exposure to environmental particles during pregnancy increases asthma susceptibility of the offspring. We tested the hypothesis that this transmission continues to F2 and F3 generations and occurs via epigenetic mechanisms. We compared allergic susceptibility of three generations of BALB/c offspring after a single maternal exposure during pregnancy to diesel exhaust particles or concentrated urban air particles. After pregnant dams received intranasal instillations of particle suspensions or control, their F1, F2, and F3 offspring were tested in a low-dose ovalbumin protocol for sensitivity to allergic asthma. We found that the elevated susceptibility after maternal exposure to particles during pregnancy persists into F2 and, with lesser magnitude, into F3 generations. This was evident from elevated eosinophil counts in bronchoalveolar lavage (BAL) fluid, histopathological changes of allergic airway disease, and increased BAL levels of IL-5 and IL-13. We have previously shown that dendritic cells (DCs) can mediate transmission of risk upon adoptive transfer. Therefore, we used an enhanced reduced representation bisulfite sequencing protocol to quantify DNA methylation in DCs from each generation. Distinct methylation changes were identified in F1, F2, and F3 DCs. The subset of altered loci shared across the three generations were not linked to known allergy genes or pathways but included a number of genes linked to chromatin modification, suggesting potential interaction with other epigenetic mechanisms (e.g., histone modifications). The data indicate that pregnancy airway exposure to diesel exhaust particles (DEP) triggers a transgenerationally transmitted asthma susceptibility and suggests a mechanistic role for epigenetic alterations in DCs in this process.

Topics: Air Pollutants; Animals; Asthma; Bronchoalveolar Lavage Fluid; Dendritic Cells; DNA Methylation; Eosinophils; Female; Interleukin-13; Interleukin-5; Maternal Exposure; Mice; Mice, Inbred BALB C; Ovalbumin; Pregnancy; Vehicle Emissions

Sophora tonkinensis (Leguminosae, ST) is a traditional herbal plant in Korea and China. Its roots and rhizomes have been used to dissipate heat, to clear toxic material and to treat acute pharyngolaryngeal infections and sore throats.. In this study, we tried to investigate the anti-inflammatory and anti-asthmatic effects of a purified extract (SKI3301) from Sophora tonkinensis using in vitro enzyme assay models and ovalbumin (OVA)-induced asthma animal models.. The effect of SKI3301 on pro-inflammatory enzymes such as 5-lipoxygenase, phosphodiesterase 3 & 4, and thromboxane synthase was assayed in vitro. BALB/c mice were sensitized with OVA/Alum ip injection and nebulized with OVA to induce airway inflammation. Bronchoalveolar lavage (BAL) fluid was collected and analyzed for leukocytes infiltration and IL-5 production along with lung histopathology. Guinea pigs passively sensitized with anti-OVA antiserum were used to investigate the effect of SKI3301 on bronchospasm in vitro and in vivo.. SKI3301 potently inhibited the activities of 5-lipoxygenase, phosphodiesterase 3 & 4, and thromboxane synthase. Orally administered SKI3301 attenuated the total leukocytes and eosinophil infiltration and IL-5 level in BAL fluids. Histopathological changes associated with lung inflammation were also reduced by SKI3301. SKI3301 inhibited OVA-induced contraction of isolated trachea from sensitized guinea pigs. SKI3301 also protected OVA-induced bronchoconstriction in the sensitized guinea pigs. Maackiain, one of 3 major components of SKI3301, was effective in inhibiting 5-lipoxygenase and OVA-induced airway inflammation.. In this study, SKI3301 potently inhibited pro-inflammatory enzymes and attenuated OVA-induced bronchospasm in animal model of allergic asthma. These results suggest that SKI3301 may have therapeutic potential for allergic asthma.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Bronchial Spasm; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Guinea Pigs; Mice; Mice, Inbred BALB C; Ovalbumin; Plant Extracts; Plant Preparations; Sophora; Trachea

Bupleurum chinense belongs to the Bupleurum spp. family that has been used in traditional herbal medicine for over thousand years. It has been reported to have anti-inflammatory, anti-oxidant, hepato-protective, antipyretic, analgesic, anti-fibrotic and immunomodulatory effect. However, the effect of B. Chinense on allergic asthma remains unclear. This study investigated the immunomodulatory effects of B. Chinense extracts (BCE) on airway inflammation in asthmatic mice model. In the ovalbumin (OVA)-induced allergic asthma model, we evaluated the number of total cells, differential inflammatory cells and the production of proinflammatory cytokines in bronchoalveolar lavage fluid (BALF) and lung homogenate as well as histological structure. The levels of NFκB p65, IκBα, p-NFκB p65, p-IκBα and the total immunoglobulin (Ig) E, anti-OVA IgE, anti-OVA IgG were also examined. The oral administration of 200mg/kg BCE inhibited the accumulation of inflammatory cells especially eosinophils in BALF. Also, BCE regulated the imbalance of Th1, Th2 and Th17-related production, with attenuated the expression of GATA3, IL-1β, IL-4, IL-5, IL-6, TNF-α and RORγt, IL-17A in BALF and lung homogenate, meanwhile, up-regulated the secretion of INF-γ in lung homogenate. The levels of IgE, anti-OVA IgE, anti-OVA IgG1 and anti-OVA IgG2a were also suppressed by BCE treatment in serum. Futhermore, BCE inhibited the proinflammatory cytokines via inactivation of NFκB p65 phosphorylation and IκBα degradation in cytoplasm. The histological analysis showed that the infiltration of inflammatory cells, mucus hypersecretion and collagen fiber deposits were ameliorated in BCE treated mice. In addition, BCE induced the functional differentiation of naive CD4+ T cells forward to Th1 and Tr1 through producing INF-γ and IL-10. These results suggest that BCE may have therapeutic potential for treating allergic asthma through inhibiting Th2/Th17 cytokines production by inactivation of NFκB pathway.

Topics: Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Bupleurum; Cytokines; Disease Models, Animal; Eosinophils; Inflammation; Lung; Male; Mice; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Plant Extracts; Signal Transduction; Th17 Cells; Th2 Cells

Isoimperatorin (IMP), an active natural furocoumarin, has numerous pharmacologic effects, including anti-inflammatory, analgesic, antispasmodic, and anticancer activities. This study aimed to evaluate the preventive activity of IMP in an ovalbumin (OVA)-induced murine model of asthma and to investigate its possible molecular mechanisms. Female BALB/c mice were sensitized on days 0 and 14 via intraperitoneal injection of 20μg OVA. On days 21-23 after the initial sensitization, the mice received an airway challenge with OVA (1% w/v in PBS) for 1h; meanwhile, IMP (10 or 30mg/kg once daily) was administered by gavage on days 18-23. Our results revealed that IMP significantly lowered the productions of interleukin (IL)-4, IL-5, IL-13, eotaxin, and immunoglobulin (Ig)E in bronchoalveolar lavage fluid (BALF), plasma, or lung tissues. Histological studies showed that IMP inhibited OVA-induced inflammatory cell infiltration and mucus production in the respiratory tract. In addition, pretreatment with IMP suppressed the activation of p38 mitogen-activated protein kinase (p38 MAPK), extracellular-signal-regulated kinases 1/2 (ERK1/2), and nuclear factor-κB (NF-κB). Together, these results suggest that IMP effectively inhibits airway inflammation and mucus hypersecretion by downregulating the levels of Th2 cytokines and inhibiting NF-κB and MAPK pathways.

Topics: Allergens; Animals; Anti-Inflammatory Agents; Asthma; Cytokines; Disease Models, Animal; Female; Furocoumarins; Humans; Immunoglobulin E; Mice; Mice, Inbred BALB C; Mitogen-Activated Protein Kinase 3; Mucus; NF-kappa B; Ovalbumin; p38 Mitogen-Activated Protein Kinases; Pneumonia; Signal Transduction; Th2 Cells

We explored the potential benefits of stigmasterol in the treatment of asthma, an airway disorder characterized by immune pathophysiology and with an ever-increasing worldwide prevalence. We assessed the modulatory effect of the intraperitoneal administration of stigmasterol on experimentally induced airway inflammation in guinea pigs. The effect of stigmasterol on inflammatory cell proliferation, oxidative stress, lung histopathology, and remodeling was investigated. The results showed significant suppressive effects on ovalbumin-induced airway inflammatory damage. Stigmasterol at 10-100 mg/kg reduced proliferation of eosinophils, lymphocytes, and monocytes while reducing peribronchiolar, perivascular, and alveolar infiltration of inflammatory cells. Histopathology revealed stigmasterol maintained lung architecture and reversed collagen deposition, an index of lung remodeling. Overexpression of serum vascular cell adhesion molecule-1 (VCAM-1) and ovalbumin-specific immunoglobulin E (OVA sIgE) elicited by ovalbumin sensitization and challenge was significantly controlled with stigmasterol. Taken together, stigmasterol possessed significant antiasthmatic properties and had suppressive effects on key features of allergen-induced asthma.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Catalase; Cell Movement; Disease Models, Animal; Eosinophils; Female; Glutathione; Guinea Pigs; Immunoglobulin E; Inflammation; Leukocyte Count; Lung; Male; Malondialdehyde; Ovalbumin; Oxidative Stress; Stigmasterol; Superoxide Dismutase; Vascular Cell Adhesion Molecule-1

Glucosamine (GlcN) is generally used as a dietary supplement because of its antiinflammatory effects. We evaluated the antiallergic effect of GlcN in mice with allergic asthma and rhinitis.. Thirty-two mice were allocated equally into 4 groups (n = 8). In group A (control), we performed intraperitoneal/intranasal challenge using sterile saline. In group B (asthma/rhinitis), we used ovalbumin for intraperitoneal/intranasal challenge to induce allergic asthma and rhinitis. In groups C and D (GlcN treatment), mice were given 1% and 5% GlcN throughout the period of ovalbumin challenge, respectively. We measured serum total and ovalbumin-specific immunoglobulin E (IgE), cytokine titers (interleukin-1, -4, -5, -6, -10, and -17; tumor necrosis factor-α; and interferon-γ), and the number of inflammatory cells (eosinophils, neutrophils, lymphocytes) in bronchoalveolar lavage (BAL) fluid. We also performed histopathologic examination of the lung and nasal cavity. Finally, we performed real-time polymerase chain reaction for the genes Bcl-2, EC-SOD, VEGF, caspase-3, Bax, COX-2, Hif-1α, and heme oxygenase-1.. Compared with group B, group D had significant serum total and ovalbumin-specific IgE decreases after GlcN treatment (p < 0.05). Titers for IL-4, IL-5, IL-6, and IL-17 in BAL fluid were significantly decreased in group D (p < 0.05). Eosinophils in BAL fluid were significantly decreased in group D compared with group B (p < 0.05). Groups C and D showed significant improvement of inflammation compared with group B. Group D had significant downregulation of EC-SOD, Bax, Hif-1α, and heme oxygenase-1 compared with group B.. GlcN had a significant antiallergic effect in mice with allergic asthma and rhinitis.

Topics: Allergens; Animals; Anti-Allergic Agents; Asthma; bcl-2-Associated X Protein; Bronchoalveolar Lavage Fluid; Cytokines; Female; Gene Expression Regulation; Glucosamine; Heme Oxygenase-1; Hypoxia-Inducible Factor 1, alpha Subunit; Immunoglobulin E; Leukocyte Count; Lung; Mice, Inbred BALB C; Nasal Mucosa; Ovalbumin; Rhinitis, Allergic; Superoxide Dismutase

Allergen-specific immunotherapy (AIT) is the only curative treatment for type-1 allergies, but sometimes shows limited therapeutic response as well as local and systemic side effects. Limited control of local inflammation and patient symptoms hampers its widespread use in severe allergic asthma.. Our aim was to evaluate whether AIT is more effective in suppression of local inflammation if performed under the umbrella of short-term non-specific immunomodulation using a small molecule inhibitor of JAK pathways.. In C57BL/6J mice, a model of ovalbumin (OVA)-induced allergic airway inflammation and allergen-specific immunotherapy was combined with the administration of Tofacitinib (TOFA, a FDA-approved JAK inhibitor) from 48 hours prior to 48 hours after therapeutic OVA-injection. The effect of TOFA on human FOXP3+CD4+ T cells was studied in vitro.. AIT combined with short-term TOFA administration was significantly more effective in suppressing total cell and eosinophil infiltration into the lung, local cytokine production including IL-1β and CXCL1 and showed a trend for the reduction of IL-4, IL-13, TNF-α and IL-6 compared to AIT alone. Furthermore, TOFA co-administration significantly reduced systemic IL-6, IL-1β and OVA-specific IgE levels and induced IgG1 to the same extent as AIT alone. Additionally, TOFA enhanced the induction of human FOXP3+CD4+ T cells.. This proof of concept study shows that JAK inhibition did not inhibit tolerance induction, but improved experimental AIT at the level of local inflammation. The improved control of local inflammation might extend the use of AIT in more severe conditions such as polyallergy, asthma and high-risk patients suffering from mastocytosis or anaphylaxis.

Topics: Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Cell Differentiation; Cytokines; Desensitization, Immunologic; Disease Models, Animal; Humans; Janus Kinases; Mice; Mice, Inbred C57BL; Ovalbumin; Piperidines; Pyrimidines; Pyrroles

Exposure to Particulate Matter (PM) could function as an adjuvant depending on the city of origin in mice allergic asthma models. Therefore, our aim was to determine whether inhalation of fine particles (PM2.5) from Mexico City could act as an adjuvant inducing allergic sensitization and/or worsening the asthmatic response in guinea pig, as a suitable model of human asthma. Experimental groups were Non-Sensitized (NS group), sensitized with Ovalbumin (OVA) plus Aluminum hydroxide (Al(OH)3) as adjuvant (S + Adj group), and sensitized (OVA) without adjuvant (S group). All the animals were exposed to Filtered Air (FA) or concentrated PM2.5 (5 h/daily/3 days), employing an aerosol concentrator system, PM2.5 composition was characterized. Lung function was evaluated by barometric plethysmography (Penh index). Inflammatory cells present in bronchoalveolar lavage were counted as well as OVA-specific IgG1 and IgE were determined by ELISA assay. Our results showed in sensitized animals without Al(OH)3, that the PM2.5 exposure (609 ± 12.73 μg/m3) acted as an adjuvant, triggering OVA-specific IgG1 and IgE concentration. Penh index increased ∼9-fold after OVA challenge in adjuvant-sensitized animals as well as in S + PM2.5 group (∼6-fold), meanwhile NS + FA and S + FA lacked response. S + Adj + PM2.5 group showed an increase significantly of eosinophils and neutrophils in bronchoalveolar lavage. PM2.5 composition was made up of inorganic elements and Polycyclic Aromatic Hydrocarbons, as well as endotoxins and β-glucan, all these components could act as adjuvant. Our study demonstrated that acute inhalation of PM2.5 acted as an adjuvant, similar to the aluminum hydroxide effect, triggering allergic asthma in a guinea pig model. Furthermore, in sensitized animals with aluminum hydroxide an enhancing influence of PM2.5 exposure was observed as specific-hyperresponsiveness to OVA challenge (quickly response) and eosinophilic and neutrophilic airway inflammation. Fine particles from Mexico City is a complex mix, which play a significant role as adjuvant in allergic asthma.

Topics: Aerosols; Air Pollutants; Allergens; Animals; Asthma; Bronchoalveolar Lavage Fluid; Guinea Pigs; Immunoglobulin E; Mexico; Mice; Mice, Inbred BALB C; Models, Animal; Ovalbumin; Particulate Matter

Lipopolysaccharide (LPS) contributes to asthma exacerbations and development of inhaled corticosteroid insensitivity. Complete resistance to systemic corticosteroids is rare, and most patients lie on a continuum of steroid responsiveness. This study aimed to examine the sensitivity of combined ovalbumin- (Ova) and LPS-induced functional and inflammatory responses to inhaled and systemic corticosteroid in conscious guinea pigs to test the hypothesis that the route of administration affects sensitivity. Guinea pigs were sensitized to Ova and challenged with inhaled Ova alone or combined with LPS. Airway function was determined by measuring specific airway conductance via whole-body plethysmography. Airway hyper-responsiveness to histamine was determined before and 24 hours post-Ova challenge. Airway inflammation and underlying mechanisms were determined from bronchoalveolar lavage cell counts and lung tissue cytokines. Vehicle or dexamethasone was administered by once-daily i.p. injection (5, 10, or 20 mg/kg) or twice-daily inhalation (4 or 20 mg/ml) for 6 days before Ova challenge or Ova with LPS. LPS exacerbated Ova-induced responses, elongating early asthmatic responses (EAR), prolonging histamine bronchoconstriction, and further elevating airway inflammation. Intraperitoneal dexamethasone (20 mg/kg) significantly reduced the elongated EAR and airway inflammation but not the increased bronchoconstriction to histamine. In contrast, inhaled dexamethasone (20 mg/ml), which inhibited responses to Ova alone, did not significantly reduce functional and inflammatory responses to combined Ova and LPS. Combined Ova and LPS-induced functional and inflammatory responses are insensitive to inhaled, but they are only partially sensitive to systemic, dexamethasone. This finding suggests that the route of corticosteroid administration may be important in determining corticosteroid sensitivity of asthmatic responses.

Topics: Administration, Inhalation; Adrenal Cortex Hormones; Animals; Asthma; Bronchial Hyperreactivity; Drug Administration Routes; Drug Combinations; Guinea Pigs; Injections, Intraperitoneal; Lipopolysaccharides; Male; Ovalbumin; Respiratory Hypersensitivity

Epidemiological evidence suggests that formaldehyde (FA) exposure may influence the prevalence and severity of allergic asthma. However, the role of genetic background in FA-induced asthma-like responses is poorly understood. In the present study, we investigated the nature and severity of asthma-like responses triggered by exposure to different doses of FA together with or without ovalbumin (OVA) in two genetically different mouse strains-BALB/c and C57BL/6. Both mouse strains were divided into two main groups: the non-sensitized group and the OVA-sensitized group. All the groups were exposed to 0, 0.5 or 3.0 mg/m3 FA for 6 h/day over 25 consecutive days. At 24 h after the final FA exposure, the pulmonary parameters were evaluated. We found that FA exposure induced Th2-type allergic responses in non-sensitized BALB/c and C57BL/6 mice. In addition, FA-induced allergic responses were significantly more prominent in BALB/c mice than in C57BL/6 mice. In sensitized BALB/c mice, however, FA exposure suppressed the development of OVA-induced allergic responses. Exposure to 3.0 mg/m3 FA in sensitized C57BL/6 mice also led to suppressed allergic responses, whereas exposure to 0.5 mg/m3 FA resulted in exacerbated allergic responses to OVA. Our findings suggest that FA exposure can induce differential airway inflammation and bronchial hyperresponsiveness in BALB/c and C57BL/6 mice.

Topics: Allergens; Animals; Asthma; Bronchial Hyperreactivity; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Formaldehyde; Humans; Inflammation; Inhalation Exposure; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Respiratory Hypersensitivity; Th2 Cells

Asthma tolerance can be induced by breast-feeding or oral feeding with ovalbumin (OVA). Anergy or deletion of specific T cells and generation of T regulatory cells might contribute to this process. However, whether respiratory syncytial virus (RSV) infection would affect asthma tolerance is not very clear. Here, we first established asthma and oral tolerance mouse models and then analyzed airway hypersensitivity and asthma-related genes in the lung, CCR6-expressing IL-17A

Topics: Animals; Asthma; Bronchoalveolar Lavage Fluid; CD3 Complex; CD4 Antigens; Cell Line, Tumor; Chemokine CCL20; Chloride Channels; Female; Immune Tolerance; Immunoglobulin E; Interleukin-17; Lung; Mice; Mice, Inbred BALB C; Mucoproteins; Ovalbumin; Receptors, CCR6; Respiratory Syncytial Virus Infections; Respiratory Syncytial Viruses

The neutrophil-activating protein of Helicobacter pylori (HP-NAP) has been identified as a modulator with anti-Th2 inflammation activity, and cholera toxin B (CTB) is a mucosal adjuvant that can also induce antigen tolerance. In this study, we constructed a CTB-NAP fusion protein on the surface of Bacillus subtilis spore and evaluate the efficiency of oral administration of the recombinant CTB-NAP spores in preventing asthma in mice. Oral administration of recombinant CTB or CTB-NAP spores significantly decreased serum ovalbumin (OVA)-specific IgE (p < 0.001) and increased fecal IgA (p < 0.01) compared to the treatment with non-recombinant spores. Oral administration of recombinant CTB or CTB-NAP spores induced IL-10 and IFN-γ expression and reduced IL-4 levels in bronchoalveolar lavage fluid (BALF). Moreover, CTB and CTB-NAP spores reduced the eosinophils in BALF and inflammatory cell infiltration in the lungs. Furthermore, CD4

Topics: Adjuvants, Immunologic; Administration, Oral; Animals; Asthma; Bacillus subtilis; Bacterial Proteins; Cholera Toxin; Hypersensitivity; Immunoglobulin E; Inflammation; Interferon-gamma; Interleukin-10; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Recombinant Fusion Proteins; Spores, Bacterial; T-Lymphocytes, Regulatory

We aimed to investigate the effect of early-life diverse microbial exposures on gut microbial colonization in an OVA-induced asthma model in BALB/c mice.. BALB/c mice were divided into 4 groups: A, offsprings were kept in a SPF environment during fetal, lactation, and childhood periods; B, offsprings were kept in the SPF environment during fetal and lactation periods, and kept in the general environment during childhood; C, offsprings were kept in the SPF environment only during fetal period, and then kept in the general environment; and D, offsprings were kept in the general environment during whole periods. The diversity of intestinal flora was analyzed using denaturing gradient gel electrophoresis. Mice were sensitized with OVA to establish an animal model of asthma. Then asthma-related inflammatory cytokines and histological analysis were performed.. The diversity of intestinal microflora in group D was significantly higher than groups A, B and C at three days and three weeks after birth, and the diversity of intestinal microflora in groups C and D were significantly higher than groups A and B at five weeks after birth. The pathologic scores of OVA-induced asthmatic mice in group D were significantly lower than group A, and serum IFN-γ levels and the IFN-γ/IL-4 ratio in group D were significantly higher than group A.. Exposure to diverse microbial environments in early life affects gut microbial colonization in BALB/c mice. The diversity of the intestinal flora in early life may prevent airway inflammation in asthma via regulating the Th1/Th2 balance.

Topics: Animals; Asthma; Bacteria; Cytokines; Disease Models, Animal; Female; Gastrointestinal Microbiome; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Phylogeny; Th1 Cells; Th2 Cells

Pulvis Fellis Suis (PFS), named with "Zhu Danfen" in China, has been extensively used for the therapy of enteritis, acute pharyngitis, whooping cough and asthma in folk medicine. Although PFS shows anti-inflammatory activities, its effect on airway inflammation in asthma has not been studied.. To explore the protective effect of PFS ethanol extract against airway inflammation in asthmatic mice.. Allergic asthma in mice was sensitized and challenged by OVA. Mice were administered in oral with PFS daily at doses of 100, 200 and 400mg/kg on days 21-27. Inflammatory cell counts and classification in bronchoalveolar lavage fluid (BALF) were analyzed. Histopathological evaluation of the lung tissue was performed by hematoxylin and eosin (H&E) and periodic acid-schiff (PAS) staining. The IgE level in serum was measured by using enzyme-linked immunoassay (ELISA). ELISA was also used to detect the levels of Th1/Th2 cytokine and eotaxin in BALF.. Histological results revealed that PFS could ameliorate OVA-induced histological changes by attenuating inflammatory cell infiltration, mucus hypersecretion and goblet cell hyperplasia in the lung. Treatment with different doses of PFS significantly decreased the elevated inflammatory cell numbers in BALF and IgE production in serum. PFS treatment reduced the production of Th2 cytokine IL-4, IL-5, IL-13, and promoted Th1 cytokine IFN-γ production in BALF. In addition, PFS also decreased the levels of eotaxin and TNF-α in BALF.. These findings suggest that PFS has a markedly anti-inflammatory effect on OVA-induced allergic asthma in mice, and could be a promising protective agent recommended for allergic asthma patients.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Female; Immunoglobulin E; Inflammation; Medicine, Chinese Traditional; Mice; Mice, Inbred ICR; Ovalbumin; Sus scrofa; Th1 Cells; Th2 Cells

Millions of people are regularly exposed to ozone, a gas known to contribute significantly to worsening the symptoms of patients with asthma. However, the mechanisms underlying these ozone exacerbation effects are not fully understood. In this study, we examined the exacerbation effect of ozone in OVA-induced asthma mice and tried to demonstrate the protective mechanism of vitamin E (VE). An asthma mouse model was established, and used to identify the exacerbating effects of ozone by assessing cytokine and serum immunoglobulin concentrations, airway leukocyte infiltration, histopathological changes in lung tissues, and airway hyper-responsiveness. We then determined the amount of reactive oxygen species (ROS) accumulated, the extent to which VE induced ROS elimination, and examined the antagonistic effects of VE on the ozone-induced exacerbating effects. This study showed that 1-ppm ozone exposure could exacerbate OVA-induced asthma in mice. More importantly we found that ozone induced oxidative stress in asthmatic airways may lead to the inhibition of Nuclear factor-erythroid 2-related factor 2 (Nrf2), and may subsequently induce even more exaggerated oxidative stress associated with asthma exacerbation. Through VE induced Nrf2 activation and the subsequent increase in Nrf2 target protein expression, this study suggests a novel mechanism for alleviating ozone exacerbated asthma symptoms.

Topics: Animals; Asthma; Humans; Male; Mice; Mice, Inbred BALB C; NF-E2-Related Factor 2; Ovalbumin; Oxidative Stress; Ozone; Reactive Oxygen Species; Vitamin E

Adenosine triphosphate (ATP) is a key mediator to alert the immune dysfunction by acting on P2 receptors. Here, we found that allergen challenge caused an increase of ATP secretion in a murine model of neutrophilic asthma, which correlated well with neutrophil counts and interleukin-17 production. When ATP signaling was blocked by intratracheal administration of the ATP receptor antagonist suramin before challenge, neutrophilic airway inflammation, airway hyperresponsiveness, and Th17-type responses were reduced significantly. Also, neutrophilic inflammation was abrogated when airway ATP levels were locally neutralized using apyrase. Furthermore, ATP promoted the Th17 polarization of splenic CD4

Topics: Adenosine Triphosphate; Allergens; Animals; Apyrase; Asthma; Cell Differentiation; Cells, Cultured; Disease Models, Animal; Female; Humans; Mice; Mice, Inbred C57BL; Mice, Transgenic; Neutrophils; Ovalbumin; Pneumonia; Purinergic P2 Receptor Antagonists; Receptors, Antigen, T-Cell, alpha-beta; Suramin; T-Lymphocyte Subsets; Th17 Cells

Viral infections are the most common triggers of asthma exacerbation, but the key molecules involved in this process have not been fully identified. Although cysteinyl leukotrienes (cysLTs) have been postulated as the key mediators, their precise roles remain largely unclear. To investigate the roles of cysLTs in virus-induced asthma exacerbation, we developed a murine model using a viral double-stranded RNA analog, polyinosinic-polycytidylic acid (poly I:C), and analyzed the effect of leukotriene receptor antagonist (LTRA) administration.. A/J mice were immunized with ovalbumin (OVA) + alum (days 0, 28, 42, and 49), followed by intranasal challenge with OVA (phase 1: days 50-52) and poly I:C (phase 2: days 53-55). Montelukast was administered during poly I:C challenge (phase 2) in the reliever model or throughout the OVA and poly I:C challenges (phases 1 and 2) in the controller model. Airway responsiveness to acetylcholine chloride was assessed, and bronchoalveolar lavage (BAL) was performed on day 56.. Administration of poly I:C to OVA-sensitized and -challenged mice increased the number of eosinophils and levels of IL-13, IL-9, CCL3, and CXCL1 in BAL fluid (BALF) and tended to increase airway responsiveness. Montelukast significantly attenuated the poly I:C-induced increase in the number of eosinophils and levels of IL-13, IL-9, and CCL3 in BALF and airway hyperresponsiveness in both the reliever and controller models.. This is the first report showing that LTRA functionally suppressed the pathophysiology of a virus-induced asthma exacerbation model, suggesting the importance of cysLTs as a potential treatment target.

Topics: Acetates; Alum Compounds; Animals; Anti-Asthmatic Agents; Asthma; Bronchoalveolar Lavage Fluid; Cyclopropanes; Cysteine; Cytokines; Disease Models, Animal; Disease Progression; Eosinophils; Immunization; Inflammation Mediators; Leukotriene Antagonists; Leukotrienes; Male; Mice; Ovalbumin; Poly I-C; Quinolines; Respiratory Hypersensitivity; RNA, Double-Stranded; RNA, Viral; Sulfides

Interleukin 33 (IL-33) represents a potential link between the airway epithelium and induction of Th2-type inflammatory responses associated with the development of asthma. This study investigated the potential of IL-33 to exacerbate antigen driven asthma responses. An ovalbumin (OVA) asthma model was used in which sensitized C57BL/6 mice were exposed to IL-33 before each OVA challenge. IL-33 given to sensitized mice acted synergistically with antigen and aggravated airway inflammation, hyperresponsiveness and remodeling compared with mice that were only OVA sensitized and challenged and mice that were only exposed to IL-33. Elevated levels of local and systemic mast cell protease mMCP-1, as well as antigen-specific IgE production, were observed following IL-33 administration to sensitized mice. Similarly, exposing OVA-sensitized mice to IL-33 increased the Th2 cytokine levels, including IL-4, IL-5 and IL-13. Furthermore, IL-33 and OVA administration to OVA-sensitized mice increased ILC2s in the lung, suggesting a role for ILC2s in IL-33-mediated exacerbation of OVA-induced airway responses. Collectively, these findings show that IL-33 aggravates important features of antigen-driven asthma, which may have implications for asthma exacerbations.

Topics: Airway Remodeling; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Inflammation; Interleukin-33; Lung; Male; Mice, Inbred C57BL; Ovalbumin; Respiratory Hypersensitivity

Asthma is an airway epithelium disorder involving allergic lung inflammation. IL-1 receptor-associated kinase M (IRAK-M) is a negative regulator of Toll-like receptor (TLR) signaling on airway epithelial cells and macrophages, and it is known to limit the overproduction of cytokines during the inflammatory process. However, the direct role of IRAK-M in asthma pathogenesis is unclear. In the present study, we found a significant elevation of IRAK-M expression in mouse lungs after ovalbumin (OVA) exposure. Compared with wild-type mice, IRAK-M knockout (KO) mice responded to OVA challenge with significantly worse infiltration of airway inflammatory cells, greater airway responsiveness, higher proinflammatory cytokine levels in lung homogenates, and more prominent T-helper cell type 2 (Th2) and Th17 deviation. OVA exposure also induced higher activities of dendritic cells (DCs) and macrophages from IRAK-M KO mouse lungs. Furthermore, adoptive transfer of either IRAK-M KO bone-marrow-derived DCs or macrophages into wild-type mice aggravated OVA-induced airway inflammation. In vitro experiments showed that IRAK-M KO naive CD4

Topics: Adoptive Transfer; Animals; Asthma; Dendritic Cells; Inflammation; Interleukin-1 Receptor-Associated Kinases; Lung; Mice; Mice, Knockout; Ovalbumin; Th2 Cells

Anti-inflammatory pharmacotherapy for asthma has mainly depended on the inhalation of glucocorticoids, which non-specifically suppress immune responses. If the anti-inflammatory cytokine interleukin (IL)-10 can be induced by a specific antigen, asthmatic airway inflammation could be suppressed when individuals are exposed to the antigen. The purpose of this study was to develop cellular immunotherapeutics for atopic diseases using IL-10-producing CD4

Topics: Adoptive Transfer; Animals; Asthma; CD4-Positive T-Lymphocytes; Cell Proliferation; Immunoglobulin E; Interleukin-10; Mice; Ovalbumin; Phenotype

Sea hare has a variety of biological activities. However, little is known regarding the anti-asthmatic effects of sea hare. This study was performed to identify the effect of sea hare hydrolysates (SHH) on an ovalbumin (OVA)-induced allergic asthma model. The experimental asthma model was sensitized and challenged with OVA. We found that a high-dose of SHH (HSHH) significantly inhibited OVA-induced airway inflammation and mucus production around the airway in lung sections, while low- and medium-dose SHH showed an insignificant effect. In addition, HSHH highly reduced OVA-induced production of interleukin-4, -5, -13, leukotriene D4, E4, and histamine in bronchoalveolar lavage fluid. HSHH decreased the histamine-induced increase in the intracellular Ca

Topics: Animal Feed; Animals; Anti-Asthmatic Agents; Asthma; Body Weight; Bronchoalveolar Lavage Fluid; Cell Survival; Dexamethasone; Female; Invertebrates; Mice; Mice, Inbred C57BL; Ovalbumin; Protein Hydrolysates; Random Allocation