epidermal-growth-factor and Inflammation

This review article intends to critically review the available literature relating to the behavior of tear-borne inflammatory biomarkers during contact lens wear.. The workflow protocol followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement recommendations. An exhaustive search was carried out using the PubMed database. The analysis included a list of 34 eligible clinical trials: Thirty addressed the use of soft contact lenses, three focused on rigid gas permeable lenses; and one on scleral lenses. The biomarkers' presence was described as changes in the molecular concentration compared to control groups - non-contact lens wearers - or baseline measurements.. Contact lens wear inflates the concentration of several inflammatory molecules in tears. Most relevant changes were found for IL-1β, IL-6, IL-8, IL-17, LTB. Mechanical trauma, hypoxia, and wearing schedules may be associated with a distinct sub-clinical inflammatory response in contact lens wearers. The relationship between these responses and contact lens-induced discomfort remains unclear, as the existing scientific evidence is still scarce. More clinical studies are still needed to prove the impact of reverse geometry and scleral lens wear on the behavior of tear-borne biomarkers.

Topics: Biomarkers; Contact Lenses, Hydrophilic; Epidermal Growth Factor; Humans; Inflammation; Interleukin-17; Interleukin-6; Interleukin-8; Matrix Metalloproteinase 9; Tears

Epithelial to mesencyhmal transition (EMT) has a central role in tumor metastasis and progression. EMT is regulated by several growth factors and pro-inflammatory cytokines. The most important role in this regulation could be attributed to transforming growth factor-β (TGF-β). In breast cancer, TGF-β effect on EMT could be potentiated by Fos-related antigen, oncogene HER2, epidermal growth factor, or mitogen-activated protein kinase kinase 5 - extracellular-regulated kinase signaling. Several microRNAs in breast cancer have a considerable role either in potentiation or in suppression of EMT thus acting as oncogenic or tumor suppressive modulators. At present, possibilities to target EMT are discussed but the results of clinical translation are still limited. In prostate cancer, many cellular events are regulated by androgenic hormones. Different experimental results on androgenic stimulation or inhibition of EMT have been reported in the literature. Thus, a possibility that androgen ablation therapy leads to EMT thus facilitating tumor progression has to be discussed. Novel therapy agents, such as the anti-diabetic drug metformin or selective estrogen receptor modulator ormeloxifene were used in pre-clinical studies to inhibit EMT in prostate cancer. Taken together, the results of pre-clinical and clinical studies in breast cancer may be helpful in the process of drug development and identify potential risk during the early stage of that process.

Topics: Benzopyrans; Breast Neoplasms; Cadherins; Cell Plasticity; Cytokines; Disease Progression; Epidermal Growth Factor; Epithelial-Mesenchymal Transition; Female; Humans; Inflammation; Male; MAP Kinase Kinase 5; Metformin; MicroRNAs; Prostatic Neoplasms; Proto-Oncogene Proteins c-fos; Receptor, ErbB-2; Signal Transduction; Transforming Growth Factor beta

Amphiregulin is a member of epidermal growth factor family, and is also one of the ligand of epidermal growth factor receptor, it participates in many physiological and pathological process by combining with EGFR. Researches have proved that AREG participates in asthma and airway inflammatory diseases caused by smoking and PM 2.5, and AREG plays an important role in the process of airway remodeling and inflammation. This paper mainly reviews the expression and function of AREG, and focus on it's research status in airway inflammatory disease.

Topics: Airway Remodeling; Amphiregulin; Asthma; Epidermal Growth Factor; ErbB Receptors; Glycoproteins; Humans; Inflammation; Intercellular Signaling Peptides and Proteins; Smoking

Pyoderma gangrenosum (PG) is a rare autoinflammatory condition in which the alteration of neutrophil function and the innate immune response play key roles in its pathogenesis. Cases of PG have been reported in patients being treated with certain medications, which may help us to understand some of the possible pathways involved in the aetiology of PG. The aim of this review is to review the cases of PG triggered by certain drugs and try to thoroughly understand the pathogenesis of the disease. To accomplish this, a PubMed search was completed using the following words: pyoderma gangrenosum, neutrophilic dermatosis, pathophysiology, drug-induced pyoderma gangrenosum. In total, we found 43 cases of drug-induced PG. Most of them were caused by colony-stimulating factors and small-molecule tyrosine kinase inhibitors. We propose that drugs induce PG through various mechanisms such as dysfunctional neutrophil migration and function, dysregulated inflammatory response, promotion of keratinocyte apoptosis and alteration of epigenetic mechanisms. PG is a rare condition with complex pathophysiology and drug-induced cases are even more scarce; this is the main limitation of this review. Understanding the possible mechanisms of drug-induced PG, via abnormal neutrophil migration and function, abnormal inflammation, keratinocyte apoptosis and alteration of epigenetic mechanisms would help to better understand the pathogenesis of PG and ultimately to optimize targeted therapy.

Topics: Apoptosis; Cell Movement; Colony-Stimulating Factors; Dermatologic Agents; Drug Eruptions; Epidermal Growth Factor; Epigenesis, Genetic; Humans; Inflammation; Keratinocytes; Neutrophils; Off-Label Use; Protein-Tyrosine Kinases; Pyoderma Gangrenosum; Tumor Necrosis Factor-alpha

Inflammation involves a series of complex biological processes mediated by innate immunity for host defense against pathogen infection. Chronic inflammation is considered to be one of the major causes of serious diseases, including a number of autoimmune/inflammatory diseases, cancers, cardiovascular diseases, and neurological diseases. Milk fat globule-epidermal growth factor 8 (MFG-E8) is a secreted protein found in vertebrates and was initially discovered as a critical component of the milk fat globule. Previously, a number of studies have reported that MFG-E8 contributes to various biological functions including the phagocytic removal of damaged and apoptotic cells from tissues, the induction of VEGF-mediated neovascularization, the maintenance of intestinal epithelial homeostasis, and the promotion of mucosal healing. Recently, emerging studies have reported that MFG-E8 plays a role in inflammatory responses and inflammatory/autoimmune diseases. This review describes the characteristics of MFG-E8-mediated signaling pathways, summarizes recent findings supporting the roles of MFG-E8 in inflammatory responses and inflammatory/autoimmune diseases, and discusses MFG-E8 targeting as a potential therapeutic strategy for the development of anti-inflammatory/autoimmune disease drugs.

Topics: Animals; Autoimmune Diseases; Epidermal Growth Factor; Glycolipids; Glycoproteins; Humans; Inflammation; Lipid Droplets; Macrophages

This review focuses on the mechanisms by which the expression of specific genes is regulated by two proteins that are important in inflammation and cancer, namely the pro-inflammatory cytokine interleukin (IL)-1β and epidermal growth factor (EGF). In the review the receptors that recognize factors that cause inflammation are described with main focus on the receptors associated with activation of IL-1β. The function of IL-1β and pathways leading to activation of transcription factors, particularly NFκB and Elk-1 are analyzed. Then the mechanisms of EGF action, with particular emphasis of the activation of Elk-1 are illustrated. The link between aberrant signaling of EGF receptor family members and cancer development is explained. The relationship between inflammation and tumorigenesis is discussed.

Topics: Animals; Epidermal Growth Factor; ErbB Receptors; Gene Expression Regulation; Humans; Inflammation; Interleukin-1; Neoplasms; Receptors, Interleukin

Mammographic density reflects variation in breast tissue composition as detected on mammogram. It is associated with a number of well-known breast cancer risk factors and itself is considered one of the strongest risk factors for breast cancer. If the expression of several proteins and genes within the breast tissue influences mammographic density in the same way as it influences breast cancer risk, then mammographic density might serve as an intermediate biomarker in future epidemiological studies on breast cancer. This has the potential to provide a quick means for predicting the effect of changes in the breast microenvironment on breast cancer risk without having to wait for an eventual development of breast cancer. In this review, the expression of several proteins and genes (growth factors, enzymes, proteoglycans and pro-inflammatory markers) within the breast tissue is shown to be associated with mammographic density. These proteins and genes are suspected to play a role in breast carcinogenesis. More studies assessing differential expression of proteins and genes in mammary epithelium and stroma and their association with mammographic density among premenopausal and postmenopausal women are required. Identification of proteins and genes influencing mammographic density may provide further insight on the molecular causes of breast cancer.

Topics: Biomarkers, Tumor; Body Mass Index; Breast Density; Breast Neoplasms; Carcinogenesis; Epidermal Growth Factor; Female; Gene Expression; Humans; Inflammation; Insulin-Like Growth Factor I; Mammary Glands, Human; Proteoglycans; Receptors, Estrogen; Receptors, Progesterone; Risk Factors

Arterial inflammation and remodeling, important sequellae of advancing age, are linked to the pathogenesis of age-associated arterial diseases e.g. hypertension, atherosclerosis, and metabolic disorders. Recently, high-throughput proteomic screening has identified milk fat globule epidermal growth factor VIII (MFG-E8) as a novel local biomarker for aging arterial walls. Additional studies have shown that MFG-E8 is also an element of the arterial inflammatory signaling network. The transcription, translation, and signaling levels of MFG-E8 are increased in aged, atherosclerotic, hypertensive, and diabetic arterial walls in vivo as well as activated vascular smooth muscle cells (VSMC) and a subset of macrophages in vitro. In VSMC, MFG-E8 increases proliferation and invasion as well as the secretion of inflammatory molecules. In endothelial cells (EC), MFG-E8 facilitates apoptosis. In addition, MFG-E8 has been found to be an essential component of the endothelial-derived microparticles that relay biosignals and modulate arterial wall phenotypes. This review mainly focuses upon the landscape of MFG-E8 expression and signaling in adverse arterial remodeling. Recent discoveries have suggested that MFG-E8 associated interventions are novel approaches for the retardation of the enhanced rates of VSMC proliferation and EC apoptosis that accompany arterial wall inflammation and remodeling during aging and age-associated arterial disease.

Topics: Animals; Apoptosis; Arteries; Endothelial Cells; Epidermal Growth Factor; Glycolipids; Glycoproteins; Humans; Inflammation; Lipid Droplets; Milk Proteins; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Signal Transduction

Wound healing is a sophisticated response ubiquitous to various traumatic stimuli leading to an anatomical/functional disruption. The aim of present article was to review the current evidence regarding the effects of microgravity on wound healing dynamics. Modulation of haemostatic phase because of alteration of platelet quantity and function seems probable. Furthermore, production of growth factors that are released from activated platelets and infiltration/function of inflammatory cells seem to be impaired by microgravity. Proliferation of damaged structures is dependent on orchestrated function of various growth factors, for example transforming growth factors, platelet-derived growth factor and epidermal growth factor, all of which are affected by microgravitational status. Moreover, gravity-induced alterations of gap junction, neural inputs, and cell populations have been reported. It may be concluded that different cellular and extracellular element involved in the healing response are modified through effect of microgravity which may lead to impairment in healing dynamics.

Topics: Epidermal Growth Factor; Evidence-Based Practice; Granulation Tissue; Hemostasis; Humans; Inflammation; Intercellular Signaling Peptides and Proteins; Platelet Activation; Platelet-Derived Growth Factor; Weightlessness; Wound Healing

Obstructive nephropathy is a major cause of renal failure, particularly in infants and children. Cellular and molecular mechanisms responsible for the progression of the tubular atrophy and interstitial fibrosis-processes that lead to nephron loss-have been elucidated in the past 5 years. Following urinary tract obstruction and tubular dilatation, a cascade of events results in upregulation of the intrarenal renin-angiotensin system, tubular apoptosis and macrophage infiltration of the interstitium. This is followed by accumulation of interstitial fibroblasts through proliferation of resident fibroblasts and epithelial-mesenchymal transformation of renal tubular cells. Under the influence of cytokines, chemokines and other signaling molecules produced by tubular and interstitial cells, fibroblasts undergo transformation to myofibroblasts that induce expansion of the extracellular matrix. The cellular interactions that regulate development of interstitial inflammation, tubular apoptosis and interstitial fibrosis are complex. Changes in renal gene expression and protein production afford many potential biomarkers of disease progression and targets for therapeutic manipulation. These include signaling molecules and receptors involved in macrophage recruitment and proliferation, tubular death signals and survival factors, and modulators of epithelial-mesenchymal transformation. Targeted gene deletion and various forms of gene therapy have been used in experimental obstructive nephropathy, mostly rodent models of unilateral ureteral obstruction or cell culture techniques. Further refinement of these models is needed to develop a matrix of biomarkers with clinical predictive value, as well as molecular therapies that will prevent or reverse the renal structural and functional consequences of obstructive nephropathy.

Topics: Apoptosis; Biomarkers; Chemokine CCL7; Epidermal Growth Factor; Fibrosis; Genetic Therapy; Humans; Inflammation; Kidney; Monocyte Chemoattractant Proteins; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Ureteral Obstruction

The integrity of oral and digestive mucosa depend on many salivary components like the Epidermal Growth Factor (EGF). Sometimes indicative, sometimes stimulated or modulated factor of oral and digestive health, EGF appears as a clinical marker in neoplastic and inflammatory diseases. As cellular growth factor, it protects the digestive mucosa with stimulation of mucus production and with inhibition of gastric secretion. Equally implicated in healing process, it enhances this one, and determines, in patients, more or less sensibility to inflammatory damages. Its strategic place in various pathologies, as stomach ulcer and tumoral process, open research prospects with a real potential of repair and pronostic.

Topics: Biomarkers; Digestive System Physiological Phenomena; Epidermal Growth Factor; Gastric Mucosa; Humans; Inflammation; Intestinal Mucosa; Mouth Mucosa; Neoplasms

Chronic obstructive pulmonary disease (COPD) is characterized by chronic obstruction of expiratory flow affecting peripheral airways, associated with chronic bronchitis (mucus hypersecretion with goblet cell and submucosal gland hyperplasia) and emphysema (destruction of airway parenchyma), together with fibrosis and tissue damage, and inflammation of the small airways. Cytokines are extracellular signalling proteins. Increased levels of interleukin (IL)-6, IL-1beta, tumour necrosis factor-alpha (TNF-alpha) and IL-8 have been measured in sputum, with further increases during exacerbations, and the bronchiolar epithelium over-expresses monocyte chemotactic protein (MCP)-1 and IL-8. IL-8 can account for some chemotactic activity of sputum, and sputum IL-8 levels correlate with airway bacterial load and blood myeloperoxidase levels. The expression of chemokines such as regulated on activation, normal T-cell expressed and secreted (RANTES) may underlie the airway eosinophilia observed in some COPD patients. Cytokines may be involved in tissue remodelling. TNF-alpha and IL-1beta stimulate macrophages to produced matrix metalloproteinase-9 (MMP-9), and bronchial epithelial cells to produce extracellular matrix glycoproteins such as tenascin. Increased expression of transforming growth factor-beta (TGFbeta) and of epidermal growth factor (EGF) occurs in the epithelium and submucosal cells of patients with chronic bronchitis. TGFbeta and EGF activate proliferation of fibroblasts, while activation of the EGF receptor leads to mucin gene expression. The cytokine profile seen in chronic obstructive pulmonary disease is different from that observed in asthma. The role of these cytokines needs to be defined and there is a potential for anticytokine therapy in chronic obstructive pulmonary disease.

Topics: Cytokines; Eosinophils; Epidermal Growth Factor; Female; Humans; Inflammation; Interleukin-1; Interleukin-8; Male; Prognosis; Pulmonary Disease, Chronic Obstructive; Sensitivity and Specificity; Severity of Illness Index; Transforming Growth Factor beta

Topics: Cell Division; Conjunctiva; Dry Eye Syndromes; Epidermal Growth Factor; Epithelial Cells; Epithelium; Humans; Inflammation; Lacrimal Apparatus; Metaplasia; Models, Biological; Surface Properties; Tears; Transforming Growth Factor beta

The healing of an adult skin lesion is a well studied but complex affair of some considerable clinical interest. Endogenous growth factors, including the EGF, FGF, PDGF and TGF beta families, are released at the wound site and presumed to be a necessary part of the natural wound healing machinery. Moreover, members of each of these families have been shown to enhance healing if added exogenously to a wound site. In this review we shall briefly discuss what is known about the mechanics and cell biology of adult wound healing, describe the normal cellular source of growth factors during the healing process and, with reference to their known capacities in tissue culture, speculate as to how particular growth factors might be able to enhance healing.

Topics: Animals; Cells, Cultured; Cicatrix; Connective Tissue; Epidermal Growth Factor; Fibrin; Fibroblast Growth Factors; Gene Expression Regulation; Growth Substances; Humans; Inflammation; Keratinocytes; Male; Mice; Platelet-Derived Growth Factor; Receptors, Cell Surface; Salivary Proteins and Peptides; Skin; Transforming Growth Factors; Wound Healing

The means by which leukocytes, including lymphocytes, monocytes, and neutrophils, migrate from the circulation to sites of acute and chronic inflammation is an area of intense research interest. Although a number of soluble mediators of these important cellular interactions have been identified, a major site of great importance to the inflammatory response is the physical interface between the white cell and the endothelium. This critical association is mediated by an array of cell surface adhesion molecules. Previous data have demonstrated that the integrin subfamily of heterotypic adhesion molecules was a major component of these adhesive interactions, although it was clear that other, non-integrin-like molecules of unknown identity also seemed to be involved during the inflammatory process. A number of these other cell-surface glycoproteins which may be involved with inflammation have recently been characterized by molecular cloning. These glycoproteins, including the peripheral lymph node homing receptor (pln HR), the endothelial cell adhesion molecule (ELAM), and PADGEM/gmp140, are all members of a family of proteins which are unified by the inclusion of three characteristic protein motifs: a lectin or carbohydrate recognition domain, an epidermal growth factor (egf) domain, and a variable number of short consensus repeats (scr) which are also found in members of the complement regulatory proteins. The appearance of lectin domains in all of these adhesion molecules is consistent with the possibility that these glycoproteins function by binding to carbohydrates which are expressed in a cell and/or region specific manner, and the members of this adhesion family have been given the generic name LEC-CAM (lectin cell adhesion molecules).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Amino Acid Sequence; Animals; Blood Platelets; Cell Adhesion Molecules; Consensus Sequence; DNA; Endothelium, Vascular; Epidermal Growth Factor; Humans; Inflammation; Lectins; Leukocytes; Membrane Glycoproteins; Mice; Molecular Sequence Data; P-Selectin; Platelet Membrane Glycoproteins; Receptors, Lymphocyte Homing; Repetitive Sequences, Nucleic Acid; Sequence Homology, Nucleic Acid

Topics: Biological Assay; Blood Platelets; Cell Differentiation; Connective Tissue Cells; Epidermal Growth Factor; Extracellular Matrix; Growth Substances; Humans; Inflammation; Insulin; Interleukin-1; Neoplasms; Nerve Growth Factors; Peptides; Platelet-Derived Growth Factor; Somatomedins

Epidermal growth factor (EGF) is one of the important peptides in wound healing process. The effects of EGF have been increasingly studied in various types of ulcers. However, data on postablative laser resurfacing wound is still limited.. To evaluate the effects of the topical EGF ointment on wound healing process and postinflammatory hyperpigmentation (PIH) prevention after fractional ablative laser resurfacing.. This is a randomized split-face study. Nineteen healthy subjects were enrolled and completed follow up protocol. Patients received single treatment of fractional carbon dioxide laser on both cheeks. After randomization, each patient was assigned to apply one side of the face with topical EGF ointment and another side with petrolatum. Wound healing was evaluated by duration of scab shedding, duration of postlaser erythema, erythema index, and transepidermal water loss on the daily follow up period of seven days after treatment. PIH was evaluated at 2, 3 weeks and 1, 2 months follow up by photographs and melanin index.. Most of patients were female with Fitzpatrick skin phototype III to V. Comparing with control (petrolatum), EGF treated side showed no significant difference in duration of scab shedding, duration of postlaser erythema, erythema index, and transepidermal water loss (P-value = .58, .22, .78, and .51, respectively). Incidence of PIH was 52.6% on EGF side and 57.9% on petrolatum side, however, it was not statistically different (P = .56). The melanin index was also not different as well (P = .96).. Topical EGF might provide significant wound healing stimulation for chronic wound more than acute wound. Further studies, especially in post laser wound or other cosmetic purposes are needed.

Topics: Adult; Cheek; Cosmetic Techniques; Epidermal Growth Factor; Erythema; Female; Humans; Hyperpigmentation; Inflammation; Lasers, Gas; Male; Ointments; Recombinant Proteins; Water Loss, Insensible; Wound Healing

Novel direct-acting antivirals (DAAs) have completely changed the panorama of hepatitis C due to their high efficacy and optimal safety profile. Unfortunately, an unexpectedly high rate of early recurrence of hepatocellular carcinoma has been reported within weeks of starting treatment, but the mechanism is not known.. We monitored the serum level of vascular endothelial growth factor (VEGF) and changes in the pattern of circulating interleukins in 103 chronic hepatitis C patients during antiviral treatment with DAA-regimens. VEGF, epidermal growth factor (EGF), and several interleukins were assessed at baseline, during treatment, and after treatment. The biological effect of DAA-treated patient serum on human umbilical vein endothelial cell (HUVEC) proliferation was also confirmed.. After 4 weeks of therapy, VEGF increased approximately 4-fold compared to baseline, remained elevated up to the end of treatment, and returned to the pre-treatment level after the end of therapy. In contrast, interleukin-10 and tumor necrosis factor-alpha significantly decreased during therapy, which was coincident with HCV clearance. The levels of both remained low after treatment. The addition of serum from patients collected during therapy induced HUVEC proliferation; however, this disappeared after the end of therapy.. DAA administration induces an early increase in serum VEGF and a change in the inflammatory pattern, coinciding with HCV clearance. This may alter the balance between inflammatory and anti-inflammatory processes and modify the antitumor surveillance of the host. Fortunately, such modifications return reverse to normal after the end of treatment.

Topics: Adult; Aged; Aged, 80 and over; Antiviral Agents; Cell Proliferation; Epidermal Growth Factor; Female; Hepacivirus; Hepatitis C; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Interleukin-10; Male; Middle Aged; Neoplasms; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

A limited number of studies have demonstrated that some modulators of inflammation can be altered by the consumption of sweet cherries. We have taken a proteomics approach to determine the effects of dietary cherries on targeted gene expression. The purpose was then to determine changes caused by cherry consumption in the plasma concentrations of multiple biomarkers for several chronic inflammatory diseases in healthy humans with modestly elevated C-reactive protein (CRP; range, 1-14 mg/L; mean, 3.5 mg/L; normal, <1.0 mg/L). Eighteen men and women (45-61 y) supplemented their diets with Bing sweet cherries (280 g/d) for 28 d. Fasting blood samples were taken before the start of consuming the cherries (study d 7), 28 d after the initiation of cherry supplementation (d 35), and 28 d after the discontinuation (d 63). Of the 89 biomarkers assessed, cherry consumption for 28 d altered concentrations of 9, did not change those of 67, and the other 13 were below the detection limits. Cherry consumption decreased (P < 0.05) plasma concentrations of extracellular newly identified ligand for the receptor for advanced glycation end products (29.0%), CRP (20.1%), ferritin (20.3%), plasminogen activator inhibitor-1 (19.9%), endothelin-1 (13.7%), epidermal growth factor (13.2%), and IL-18 (8.1%) and increased that of IL-1 receptor antagonist (27.9%) compared with corresponding values on study d 7. The ferritin concentration continued to decrease between d 35 and 63 and it was significantly lower on d 63 than on d 7. Because the participants in this study were healthy, no clinical pathology end points were measured. However, results from the present study demonstrate that cherry consumption selectively reduced several biomarkers associated with inflammatory diseases.

Topics: Biomarkers; C-Reactive Protein; Chronic Disease; Diet; Dietary Supplements; Endothelin-1; Epidermal Growth Factor; Female; Ferritins; Fruit; Humans; Inflammation; Inflammation Mediators; Interleukin-18; Male; Middle Aged; Phytotherapy; Plant Preparations; Plasminogen Activator Inhibitor 1; Proteomics; Prunus; Receptors, Interleukin-1; Reference Values

Chronic obstructive pulmonary disease (COPD) is characterized by progressive worsening of airflow limitation associated with abnormally inflamed airways in older smokers. Despite correlative evidence for a role for tumor necrosis factor-alpha in the pathogenesis of COPD, the anti-tumor necrosis factor-alpha, infliximab did not show clinical efficacy in a double-blind, placebo-controlled, phase II clinical trial. This study sought to evaluate the systemic inflammatory profile associated with COPD and to assess the impact of tumor necrosis factor neutralization on systemic inflammation.. Serum samples (n = 234) from the phase II trial were collected at baseline and after 24 weeks of placebo or infliximab. Additionally, baseline serum samples were obtained from an independent COPD cohort (n = 160) and 2 healthy control cohorts (n = 50; n = 109). Serum concentrations of a broad panel of inflammation-associated analytes were measured using a 92-analyte multiplex assay.. Twenty-five proteins were significantly elevated and 2 were decreased in COPD, including highly elevated CD40 ligand, brain-derived neurotrophic factor, epidermal growth factor, acute-phase proteins, and neutrophil-associated proteins. This profile was largely independent of smoking status, age, and clinical phenotype. The majority of these associations of serum analytes with COPD are novel findings. Increased serum creatine kinase-muscle/brain and myoglobin correlated modestly with decreased forced expiratory volume at 1 second, suggesting cardiac involvement. Infliximab did not affect this systemic inflammatory profile.. A robust systemic inflammatory profile was associated with COPD. This profile was generally independent of disease severity. Because anti-tumor necrosis factor-alpha did not influence systemic inflammation, how to control the underlying pathology beyond symptom suppression remains unclear.

Topics: Acute-Phase Proteins; Adult; Aged; Anti-Inflammatory Agents; Antibodies, Monoclonal; Brain-Derived Neurotrophic Factor; C-Reactive Protein; CD40 Antigens; Creatine Kinase; Epidermal Growth Factor; Female; Humans; Inflammation; Infliximab; Male; Middle Aged; Pulmonary Disease, Chronic Obstructive; Severity of Illness Index; Tumor Necrosis Factor-alpha

OBJECTIVE-To characterize healing of corneal epithelial defects in horses and to evaluate the ability of epidermal growth factor (EGF) to modulate rate of corneal epithelial healing in horses.. 20 eyes in 12 adult horses.. Corneal epithelial wounds were created by mechanically debriding the limbus. Corneal healing was recorded for 3 treatment groups: 50 microg of EGF/ml (n = 5 eyes), 5 microg of EGF/ml (7), and PBS solution (8). Corneal healing was recorded once daily after instillation of fluorescein stain by use of photography and calculating the area of the wound, using imaging software.. After corneal debridement, re-epithelialization was rapid and progressed in a linear fashion for the first 5 to 7 days after surgery in all groups. After that period, rates of healing decreased. A profound increase in the degree of inflammation, neovascularization, melanosis, and scarring was observed in eyes treated with the high dose of EGF (50 microg/ml), but there was not a statistical difference in mean healing time or in mean decrease in radius during the linear phase between the control and either EGF treatment groups. However, for all 8 horses in which both eyes were debrided, the first eye healed significantly faster than the second eye, regardless of treatment.. Beneficial effects of topical administration of a high dose of EGF for acceleration of healing of corneal defects in eyes of horses are outweighed by the intensity of the associated inflammatory response.

Topics: Administration, Topical; Animals; Corneal Diseases; Epidermal Growth Factor; Epithelial Cells; Epithelium, Corneal; Female; Horse Diseases; Horses; Humans; Inflammation; Male; Wound Healing

Treatment of hypertrophic scars can be difficult for both patients and physicians. Silicone-containing gel dressings have been reported to be an effective alternative treatment for hypertrophic scars, yet the mechanism of action of these dressings is unknown.. To determine whether silicone is an essential factor in the treatment of hypertrophic scars and investigate the effects of occlusive dressing therapy on the expression of key wound healing mediators.. A pilot paired comparison, nonrandomized study was conducted comparing a silicone gel sheeting (Silastic [SGS]) with a hydrogel dressing (ClearSite). The effects of the dressings were compared side by side in the treatment of 15 hypertrophic scars at both the clinical and molecular levels through the use of reverse transcriptase/polymerase chain reaction to evaluate effects on the expression of interleukin 8 (IL-8), basic fibroblast growth factor (bFGF), granulocyte-macrophage colony-stimulating factor (GMCSF), epidermal growth factor (EGF), transforming growth factor beta (TGF-beta), and fibronectin.. Comparable clinical improvement of the hypertrophic scars was obtained with both dressings. Treatment of hypertrophic scars resulted in increased mean levels of IL-8, bFGF, and GMCSF mRNA; while mean TGF beta and fibronectin mRNAs decreased after treatment with both dressings. Comparison between the two dressings revealed significant changes in IL-8 and fibronectin mRNA levels after treatment with ClearSite, while only fibronectin changes were significant after treatment with SGS with respect to normal skin. Only ClearSite induced significant changes in IL-8 and bFGF levels when untreated scars were compared with posttreatment lesions, suggesting that the hydrogel augments collagenolysis via promotion of inflammation.. This study demonstrates that silicone is not a necessary component of occlusive dressings in the treatment of hypertrophic scars. The pathogenesis of hypertrophic scars is further elucidated by demonstrating that there is molecular evidence for extensive connective tissue remodeling occurring during occlusive dressing therapy.

Topics: Adult; Aged; Cicatrix, Hypertrophic; Collagen; Connective Tissue; Cytokines; Epidermal Growth Factor; Fibroblast Growth Factor 2; Fibronectins; Gels; Gene Expression Regulation; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation; Interleukin-8; Middle Aged; Occlusive Dressings; Pilot Projects; Polyethylene Glycols; Polymerase Chain Reaction; Polyurethanes; RNA, Messenger; Silicone Elastomers; Silicones; Skin; Transforming Growth Factor beta; Wound Healing

The Notch signaling pathway is an important therapeutic target for the treatment of inflammatory diseases and cancer. We previously created ligand-specific inhibitors of Notch signaling comprised of Fc fusions to specific EGF-like repeats of the Notch1 extracellular domain, called Notch decoys, which bound ligands, blocked Notch signaling, and showed anti-tumor activity with low toxicity. However, the study of their function depended on virally mediated expression, which precluded dosage control and limited clinical applicability. We have refined the decoy design to create peptibody-based Notch inhibitors comprising the core binding domains, EGF-like repeats 10-14, of either Notch1 or Notch4. These Notch peptibodies showed high secretion properties and production yields that were improved by nearly 100-fold compared to previous Notch decoys. Using surface plasmon resonance spectroscopy coupled with co-immunoprecipitation assays, we observed that Notch1 and Notch4 peptibodies demonstrate strong but distinct binding properties to Notch ligands DLL4 and JAG1. Both Notch1 and Notch4 peptibodies interfere with Notch signaling in endothelial cells and reduce expression of canonical Notch targets after treatment. While prior DLL4 inhibitors cause hyper-sprouting, the Notch1 peptibody reduced angiogenesis in a 3-dimensional in vitro sprouting assay. Administration of Notch1 peptibodies to neonate mice resulted in reduced radial outgrowth of retinal vasculature, confirming anti-angiogenic properties. We conclude that purified Notch peptibodies comprising EGF-like repeats 10-14 bind to both DLL4 and JAG1 ligands and exhibit anti-angiogenic properties. Based on their secretion profile, unique Notch inhibitory activities, and anti-angiogenic properties, Notch peptibodies present new opportunities for therapeutic Notch inhibition.

Topics: Angiogenesis Inhibitors; Animals; Endothelial Cells; Epidermal Growth Factor; Immunoprecipitation; Inflammation; Ligands; Mice; Neoplasms; Receptor, Notch1; Receptor, Notch4; Recombinant Fusion Proteins; Retinal Vessels; Surface Plasmon Resonance

This experimental study evaluated the anti-asthmatic potential of the Rho-kinase inhibitor hydroxyfasudil in the settings of allergen-induced allergen-induced experimental asthma.. Chronic allergic airway inflammation was caused by 28 days-sensitisation of guinea pigs with ovalbumin (OVA). Hydroxyfasudil was administered intraperitoneally in two doses for the last two weeks (1 mg/kg b.w.; 10 mg/kg b.w.). The degree of allergic inflammation was determined based on concentrations of inflammatory Th2 cytokines (IL-4, IL-13), Th1 cytokines (TNF-α and IFN-γ) in the lung homogenate and leukocyte count in the bronchoalveolar lavage fluid (BALF). The markers of remodelling and fibrosis, the growth factors (TGF-β1, EGF), EGF receptor, collagen type III and V were estimated in lung homogenate. The changes in specific airway resistance (sRaw) were used as an in vivo bronchial hyperreactivity parameter.. Hydroxyfasudil administration at both doses significantly reduced sRaw after a week of therapy. We observed a decline of IL-13, TNF-α and IFN-γ in lung homogenate and a lower presence of lymphocytes in BALF after 14 days of hydroxyfasudil administration at both tested doses. Hydroxyfasudil 14 days-treatment at both doses effectively reduced the concentrations of TGF-β1, EGF receptors, collagen type III and V in BALF and modulated EGF levels.. These findings indicate that RhoA/Rho-kinase is involved in the pathophysiology of allergic airway inflammation and suggest that Rho-kinase inhibitor hydroxyfasudil has therapeutic potential for asthma management.

Topics: Allergens; Animals; Anti-Asthmatic Agents; Bronchoalveolar Lavage Fluid; Cytokines; Disease Models, Animal; Epidermal Growth Factor; Guinea Pigs; Inflammation; Interleukin-13; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; rho-Associated Kinases; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha

Activity and chronicity of kidney involvement in ANCA-associated vasculitis (AAV) can be currently reliably evaluated only by kidney biopsy. In this study, we measured a panel of serum and urinary biomarkers collected at the time of kidney biopsy and hypothesized that they could reflect specific histopathological parameters in the biopsy and help to predict prognosis.. We examined a cohort of 45 patients with AAV and 10 healthy controls. Biomarker levels (DKK-3, CD163, EGF, PRO-C6 and C3M) were measured in this study by ELISA. Biopsies were scored with a scoring system for AAV (focal x crescentic x sclerotic x mixed class) and interstitial fibrosis was quantified.. Levels of urinary DKK-3, CD163, EGF, PRO-C6 and C3M significantly differed among biopsy classes in AAV, with urinary DKK-3 and PRO-C6 levels being highest in the sclerotic class and lowest in the focal class, urinary CD163 levels highest in the crescentic class and urinary C3M levels highest in the focal class. Moreover, the urinary biomarkers were able to discriminate focal biopsy class from the other classes. Urinary DKK-3, EGF, PRO-C6 and C3M levels measured at the time of biopsy were also significantly related to the extent of fibrosis and to the final kidney function at the end of follow-up.. This small pilot study suggests that selected urinary biomarkers of fibrosis and inflammation may reflect changes in the kidney biopsy and be prognostic of kidney outcome in patients with AAV.

Topics: Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis; Antibodies, Antineutrophil Cytoplasmic; Biomarkers; Epidermal Growth Factor; Fibrosis; Humans; Inflammation; Kidney; Pilot Projects

Chronic nonhealing diabetic wounds are a serious complication of diabetes, with a high morbidity rate that can cause disability or death. The long period of inflammation and dysfunctional angiogenesis are the main reasons for wound-healing difficulty in diabetes. In this study, a multifunctional double-layer microneedle (DMN) is constructed to control infection and promote angiogenesis, meeting the multiple demands of the healing process of a diabetic wound. The double-layer microneedle is consisted in a hyaluronic acid substrate and a mixture of carboxymethyl chitosan and gelatin as the tip. The antibacterial drug tetracycline hydrochloride (TH) is loaded into the substrate of the microneedle to achieve rapid sterilization and promote resistance to external bacterial infections. The microneedle tip loaded with recombinant human epidermal growth factor (rh-EGF) is inserted into the skin, in response to gelatinase produced by resident microbe and disassociate to achieve the enzymatic response release. The double-layer drug-loaded microneedles (DMN@TH/rh-EGF) have antibacterial and antioxidant effects, and promote cell migration and angiogenesis in vitro. In an in vivo diabetic wound model, using rats, the DMN@TH/rh-EGF patch is able to inhibit inflammation, promote angiogenesis, collagen deposition, and tissue regeneration during the wound healing process, promoting its healing.

Topics: Animals; Anti-Bacterial Agents; Diabetes Mellitus; Epidermal Growth Factor; Humans; Inflammation; Rats; Wound Healing

Salidroside has been widely used in anti-tumor, cardiovascular, and cerebrovascular protection. However, there are few reports of its use for wound repair. Herein, salidroside inflammation-targeted emulsion gel and non-targeted emulsion gel were developed for wound repair. The inflammation-targeted emulsion gels showed an overall trend of better transdermal penetration and lower potential than non-targeted emulsion gels (-58.7 mV and -1.6 mV, respectively). The apparent improvement of the trauma surface was significant in each administration group. There was a significant difference in the rate of wound healing of the rats between each administration group and the model group at days 7 and 14. Pathological tissue sections showed that inflammatory cells in the epidermis, dermis, and basal layer were significantly reduced, and the granulation tissue was proliferated in the inflammation-targeted emulsion gel group and the non-targeted emulsion gel group. Regarding the expressions of EGF and bFGF, the expressions of bFGF and EGF in the tissues of the inflammation-targeted group at days 7, 14, or 21 were significantly higher than that of the non-targeted emulsion gel group and the model group, both of which were statistically significant compared with the model group (

Topics: Animals; Emulsions; Epidermal Growth Factor; Gels; Inflammation; Rats; Wound Healing

Psoriasis is a chronic inflammatory skin disease in which growth activity is more prominent than inflammatory activity at the centre of lesional skin (CE skin). This growth activity is partly influenced by growth factors (GFs) that play an important role in cell growth and inflammation during the plaque development. In this study, we identified potential GFs in CE skin and predicted their regulatory functions and biological activity in mediating transcripts in the plaques. Samples of uninvolved skin (UN skin) and CE skin were biopsied from patients with psoriasis vulgaris for RNA-sequencing analysis in order to identify differentially expressed genes (DEGs). Our finding revealed that epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF) and hepatocyte growth factor (HGF) signalling were enriched by CE/UN skin-derived DEGs. Additionally, several EGFR ligands, namely EGF, heparin-binding EGF like growth factor (HB-EGF), amphiregulin (AREG) and transforming growth factor (TGF)-α, as well as TGF-β1, TGF-β2, vascular endothelial growth factor-A, FGFs, PDGF-B and HGF, were predicted to be GF regulators. The regulatory pattern and biological activity of these GF regulators on mediating the CE/UN skin-derived DEGs was demonstrated. This study provides a novel hypothesis regarding the overall regulatory function of GFs, which appear to modulate the expression of the transcripts involved in inflammation and growth in the CE skin. In addition, some GFs may exert anti-inflammatory effects. Further investigations on the mechanisms underlying this regulation may contribute to a deeper understanding of psoriasis and the identification of potential therapeutic targets for patients with psoriasis.

Topics: Epidermal Growth Factor; Fibroblast Growth Factors; Humans; Inflammation; Psoriasis; Skin; Vascular Endothelial Growth Factor A

Polyphenols have been widely used to treat various chronic skin diseases because they are beneficial in wound healing and show anti-inflammatory effects, however, the mechanism of action remains ambiguous. Previously, we reported the wound healing capability of tea polyphenols (TPP), the major functional component of tea, in vivo. The current study aimed to address the mechanisms of TPP in wound healing during different phases (inflammation, proliferation and remodeling). During the inflammation phase, TPP reduced the production of proinflammatory cytokines (IL-1β, IL-6 and TNF-α) and inhibited infiltration of neutrophils; during the proliferation phase, TPP promoted the expression of growth factor VEGF-A, which can promote vascular endothelial cell division and induce angiogenesis; TPP improved the morphology of the wound and restored the ratio of type III/I collagens during the remodeling phase, as determined by Masson-trichrome staining and Sirius red staining assays. By tracking the changes in the wound area, TPP and recombinant human epidermal growth factor (rhEGF), rather than povidone-iodine (PVP-I), were able to promote wound healing. These results suggest that TPP plays a pivotal role in all the key stages of wound healing and displays distinct mechanisms from rhEGF, suggesting clinical significance for the future application of TPP as a natural wound healing agent.

Topics: Animals; Biological Assay; Clinical Relevance; Collagen Type I; Collagen Type III; Disease Models, Animal; Epidermal Growth Factor; Humans; Inflammation; Mice; Tea

Cardiovascular (CV) morbidity and mortality, and perpetuated synovial angiogenesis have been associated with RA. In our study we evaluated angiogenic factors in relation to vascular inflammation and function, and clinical markers in RA patients undergoing 1-year tofacitinib therapy.. Thirty RA patients treated with either 5 mg or 10 mg twice daily tofacitinib were included in a 12-month follow-up study. Eventually, 26 patients completed the study and were included in data analysis. Levels of various angiogenic cytokines (TNF-α, IL-6), growth factors [VEGF, basic fibroblast (bFGF), epidermal (EGF), placental (PlGF)], cathepsin K (CathK), CXC chemokine ligand 8 (CXCL8), galectin-3 (Gal-3) and N-terminal prohormone brain natriuretic peptide (NT-proBNP) were determined at baseline, and at 6 and 12 months after initiating tofacitinib treatment. In order to assess flow-mediated vasodilation, common carotid intima-media thickness (ccIMT) and carotid-femoral pulse-wave velocity, ultrasonography was performed. Synovial and aortic inflammation was also assessed by 18F-fluorodeoxyglucose-PET/CT.. One-year tofacitinib therapy significantly decreased IL-6, VEGF, bFGF, EGF, PlGF and CathK, while it increased Gal-3 production (P < 0.05). bFGF, PlGF and NT-proBNP levels were higher, while platelet-endothelial cell adhesion molecule 1 (PECAM-1) levels were lower in RF-seropositive patients (P < 0.05). TNF-α, bFGF and PlGF correlated with post-treatment synovial inflammation, while aortic inflammation was rather dependent on IL-6 and PECAM-1 as determined by PET/CT (P < 0.05). In the correlation analyses, NT-proBNP, CXCL8 and Cath variables correlated with ccIMT (P < 0.05).. Decreasing production of bFGF, PlGF or IL-6 by 1-year tofacitinib therapy potentially inhibits synovial and aortic inflammation. Although NT-proBNP, CXCL8 and CathK were associated with ccIMT, their role in RA-associated atherosclerosis needs to be further evaluated.

Topics: Arthritis, Rheumatoid; Biomarkers; Carotid Intima-Media Thickness; Epidermal Growth Factor; Female; Follow-Up Studies; Humans; Inflammation; Interleukin-6; Placenta; Platelet Endothelial Cell Adhesion Molecule-1; Positron Emission Tomography Computed Tomography; Pregnancy; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Topics: Animals; Collagen; Collagen Type I; Eels; Epidermal Growth Factor; Inflammation; Interleukins; Mice; Pepsin A; Skin; Tumor Necrosis Factor-alpha; Urinary Bladder; Wound Healing

To determine the effect of sodium hyaluronate combined with recombinant human epidermal growth factor (rhEGF) on clinical symptoms and inflammation in patients with newly diagnosed xerophthalmia after cataract surgery.. A total of 106 patients who underwent cataract surgery and were newly diagnosed with xerophthalmia in our hospital between June 2018 and August 2019 were enrolled. Of these, 50 patients who were treated with sodium hyaluronate (0.1%) were assigned to the monotherapy group (MG) and the remaining 56 patients who were treated with sodium hyaluronate (0.1%) combined with rhEGF (20 μg/ml) were assigned to the combination group (CG). The 2 groups were compared based on ocular surface disease index (OSDI) score, break-up time (BUT), fluorescein corneal staining level, Schirmer I test (SI) level, clinical efficacy (disappearance of typical symptoms, including eyes drying, burning sensation, foreign body sensation, etc), and interleukin (IL)-1, IL-6, and tumor necrosis factor-α (TNF-α) levels. Spearman correlation analysis was conducted to analyze the relationship between IL-1, IL-6, TNF-α and clinical efficacy. In addition, receiver operating characteristic curves were drawn to analyze the predictive value of IL-1, IL-6, and TNF-α in efficacy on xerophthalmia.. After treatment, the CG showed reduced OSDI score compared with the MG. The CG showed increased BUT (s) and SI (mm) levels compared with MG. After treatment, the CG exhibited decreased levels of IL-1(ng/mL), IL-6 (ng/mL), and TNF-α (ng/mL) compared with the MG. Spearman correlation analysis revealed that IL-1, IL-6, and TNF-α were negatively correlated with clinical efficacy. The areas under the curves of IL-1, IL-6, and TNF-α were 0.801, 0.800, and 0.736 respectively.. Sodium hyaluronate combined with rhEGF is helpful to alleviate clinical symptoms and inflammation in patients with xerophthalmia undergoing cataract surgery.

Topics: Cataract; Cataract Extraction; Epidermal Growth Factor; Humans; Hyaluronic Acid; Inflammation; Ophthalmic Solutions; Recombinant Proteins; Xerophthalmia

The present study evaluated the antiparasitic effect of curcumin extract on Schistosoma mansoni in Swiss albino mice. The experimental design included four groups of S. mansoni - infected mice; without treatment (controls), curcumin-treated, Praziquantel (PZQ)-treated, and PZQ +curcumin treated mice. The results showed that curcumin improved ISHAK confluent necrosis score up to zero. PZQ +curcumin showed a significant reduction in portal inflammation. Both activity and fibrosis demonstrated lower scores in all treated groups, however, PZQ revealed a marked increase in confluent necrosis and interface hepatitis. Besides, the lobular inflammation revealed worsening in the overall ISHAK score in all treated groups compared with the control. Few periocular granulomas were recovered by PZQ +curcumin treatment at day 35 post-treatment (6±1.2), P-value <0.05. Curcumin revealed a mild reduction (60±7.376). Curcumin-treated groups, with and without PZQ, resulted in higher significant Immunoreactivity score (IRS) for Bcl-2-associated X (BAX) and lower Interleukine- 17A (IL-17A), and Human epidermal growth factor (EGF), compared to the control. However, PZQ revealed a lower mean IRS value in BAX, higher IL-17A and EGF in the periovulatory granuloma. It was concluded that PZQ +curcumin treatment had a potent synergistic outcome through lessening the number of granulomas, the inflammatory events, and the expression of EGF, and amelioration of apoptosis in the periovulatory granulomas if compared with either PZQ or curcumin alone.

Topics: Animals; Anthelmintics; bcl-2-Associated X Protein; Curcumin; Epidermal Growth Factor; Granuloma; Inflammation; Interleukin-17; Mice; Necrosis; Praziquantel; Schistosomiasis mansoni

A high concentration of blood ammonia can lead to failure of early embryo implantation in dairy cows. Progesterone (P4) is an important hormone that regulates the changes of uterine receptivity and participates in pregnancy during early implantation. However, whether P4 alleviates the harmful effects of blood ammonia is unknown. Therefore, this experiment treated bovine endometrial epithelial cells (BEEC) with ammonia chloride and P4. The results showed that ammonia decreased the viability of BEEC, upregulated the ROS level and apoptosis rate, and upregulated the expressions of Bax, Bcl-2, P53, Cyt-c, caspase-3, caspase-8 and caspase-9. It also upregulated the expressions of TLR4, NF-κB, and in turn, produced a large amount of IL-6 and downregulated the expressions of EGF and VEGF. After co-treating BEEC with ammonia and P4, P4 had no alleviating effect on cell viability, ROS and apoptosis, but downregulated the expressions of TLR4, NF-κB, IL-6, and upregulated the expressions of LIF, EGF and VEGF. Inhibition of STAT3 signaling pathway, LIF expression was not affected, and VEGF expression was down-regulated. Using LIF treated BEEC, VEGF expression was upregulated, but after the addition of inhibitors, the expression of VEGF was not upregulated by LIF. Therefore, progesterone could not alleviate the oxidative stress and apoptosis induced by ammonia in BEEC. However, progesterone ameliorated the inflammation and receptivity genes of BEEC induced by ammonia via the NF-κB and LIF/STAT3 signaling pathways.

Topics: Ammonia; Animals; Cattle; Cattle Diseases; Embryo Implantation; Epidermal Growth Factor; Female; Inflammation; Interleukin-6; NF-kappa B; Pregnancy; Progesterone; Reactive Oxygen Species; Toll-Like Receptor 4; Vascular Endothelial Growth Factor A

Epidermal growth factor (EGF) has many important biological functions. It plays an important role in regulating the growth, survival, migration, apoptosis, proliferation, and differentiation of intestinal tissues and cells. However, until now, the effect of inflammation on the biological activity of EGF in intestinal cells or tissues is still unclear. For this reason, in the current research, we have conducted a detailed study on this issue. Using the rat small intestinal crypt epithelial cell line (IEC6) was used as an in vitro model, and Confocal laser scanning microscope (CLSM), Flow cytometry (FCM), Indirect immunofluorescence assay (IFA), Western-blotting (WB), and Quantitative real-time RT-PCR (QRT-PCR) methods were used to explore the effects of inflammation on EGF/EGFR biological activity and signal transduction profiles. We found that the EGF/EGFR nuclear signal almost disappeared in the inflammatory state, and the phosphorylation levels of EGFR, AKT, and STAT3 were all significantly down-regulated. In addition, we also studied the effect of β-carotene on the biological activity of EGF, and found that when cells were pretreated with β-carotene, the cellular behavior, biological activity, and nuclear signal of EGF/EGFR under inflammation stimulation were partially restored. In summary, the current study shows that inflammation can disrupt EGF/EGFR-mediated signaling in IEC6 cells, suggesting that inflammation negatively regulates the biological activity of EGF/EGFR. Furthermore, we found that β-carotene not only attenuated lipopolysaccharide (LPS)-induced inflammation but also partially restored the biological activity of EGF in IEC6 cells, laying a solid foundation for studying the biological functions of EGF and β-carotene.

Topics: Animals; beta Carotene; Cell Line, Tumor; Epidermal Growth Factor; ErbB Receptors; Inflammation; Phosphorylation; Rats; Signal Transduction

To investigate the effects of local epidermal growth factor (EGF) use on anastomotic healing during primary repair of anastomosis in rats with anastomotic leaks (AL).. Thirty albino Wistar rats were divided into three groups. Anastomoses were performed in group 1 after colon transection. In groups 2 and 3, ALs were created with an incomplete colon anastomosis model. Relaparotomy was conducted on rats in groups 2 and 3 72 h after the first procedure. ALs of the rats were repaired with a primary suture in group 2 and with a primary suture and the application of submucosal EGF in group 3. All rats were sacrificed through cervical dislocation on the 6th day after the first procedure. Four-centimeter colonic segments containing 2-cm distal and proximal parts of the anastomotic lines of the subjects were resected. The primary outcome was anastomotic burst pressure (ABP). The secondary outcomes included limitation in inflammation, increased neovascularization, increased fibroblast activation and increased collagen synthesis.. The ABP value of group 2 was significantly lower than that of group 3 (P < 0.05). No significant difference was detected in the ABP value between group 3 and group 1 (P > 0.05). There was significantly less inflammatory cell infiltration in group 3 than in group 2 (P < 0.05). Collagen synthesis and neovascularization were significantly higher in group 3 than in group 2 (P < 0.05).. A single-dose of submucosal EGF applied to the AL line limited inflammation and stimulated neovascularization. It also had a positive effect on the strength of the anastomosis.

Topics: Anastomosis, Surgical; Anastomotic Leak; Animals; Collagen; Colon; Epidermal Growth Factor; Humans; Inflammation; Rats; Rats, Wistar

Idiopathic pulmonary fibrosis (IPF) is a poorly understood, progressive lethal lung disease with no known cure. In addition to alveolar epithelial cell (AEC) injury and excessive deposition of extracellular matrix proteins, chronic inflammation is a hallmark of IPF. Literature suggests that the persistent inflammation seen in IPF primarily consists of monocytes and macrophages. Recent work demonstrates that monocyte-derived alveolar macrophages (moAMs) drive lung fibrosis, but further characterization of critical moAM cell attributes is necessary. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is an important epidermal growth factor receptor ligand that has essential roles in angiogenesis, wound healing, keratinocyte migration, and epithelial-mesenchymal transition. Our past work has shown HB-EGF is a primary marker of profibrotic M2 macrophages, and this study seeks to characterize myeloid-derived HB-EGF and its primary mechanism of action in bleomycin-induced lung fibrosis using

Topics: Animals; Bleomycin; Epidermal Growth Factor; Heparin; Heparin-binding EGF-like Growth Factor; Humans; Idiopathic Pulmonary Fibrosis; Inflammation; Mice

To determine the clinical efficacy of recombinant human epidermal growth factor (rh-EGF) combined with povidone-iodine (PVI) on patients with pressure ulcers (PUs).. One hundred and five PU patients treated between January 2018 and January 2021 were enrolled and retrospectively analyzed. Of them, 50 patients who received conventional treatment were assigned to the control group (Con group), while 55 patients treated with rh-EGF combined with PVI were assigned to the observation group (Obs group). The two groups were compared in clinical efficacy, PU alleviation (total area reduction rate, total depth reduction rate, and total volume reduction rate), healing time, pain degree (Visual Analog Scale [VAS] score), inflammatory indexes (interleukin-8 [IL-8], tumor necrosis factor-. The Obs group yielded a higher total effective rate than the Con group (. All in all, rh-EGF combined with PVI has a definite curative effect on patients with PUs. It can promote PU alleviation and hydroxyproline secretion in the wound and inhibit pain and inflammatory reactions, which is worthy of clinical promotion.

Topics: Cytokines; Epidermal Growth Factor; Humans; Hydroxyproline; Immunologic Factors; Inflammation; Pain; Povidone-Iodine; Pressure Ulcer; Retrospective Studies; Suppuration

Peptic ulcer episodes cause damage to the stomach and intestine, with inflammatory cell infiltration and oxidative stress as the main players. In this study, we investigated the potential of anthocyanidin malvidin for preventive and curative peptic ulcer treatment. The anthocyanidin effects were examined in gastric ulcer mouse models induced by ethanol, non-steroidal anti-inflammatory drugs (NSAIDs), ischemia-reperfusion (IR), acetic acid and duodenal ulcer induced by polypharmacy. Expression levels of oxidative and inflammatory genes were measured to investigate the mechanism of anthocyanin activity. At a dose of 5 mg·kg

Topics: Acetic Acid; Animals; Anthocyanins; Antioxidants; Biomarkers; Cyclooxygenase 1; Disease Models, Animal; Duodenum; Epidermal Growth Factor; Ethanol; Gastric Mucosa; Gene Expression Regulation; Indomethacin; Inflammation; Male; Matrix Metalloproteinase 9; Mice; Oxidative Stress; Peptic Ulcer; Polypharmacy; Protective Agents; Reperfusion Injury; Stomach Ulcer; Tight Junctions; Wound Healing

BACKGROUND The aim of this study was to determine the effect of kangfuxin liquid (KFXL) on inflammatory response, and its underlying mechanism in treating acute ulcerative colitis (UC) in mice induced by dextran sulfate sodium (DSS). MATERIAL AND METHODS Mice were provided drinking water containing DSS (3%) for 7 days to induce acute enteritis. The mice were divided into 6 groups: a control group, a DSS-induced (vehicle) group, a sulfasalazine (SASP) group, and low-, medium-, and high-dose kangfuxin liquid groups. Disease activity index (DAI), colon mucosa damage index (CMDI), histopathological score (HS), and organ index were monitored daily. The levels of interleukin-1ß (IL-1ß), interleukin-10 (IL-10) in serum and interleukin-17 (IL-17) and epidermal growth factor (EGF) in colon tissue were assessed by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was used to assess the changes of T lymphocyte subsets in spleens of mice to evaluate the therapeutic effect of drugs on acute UC in mice. RESULTS Different doses of kangfuxin liquid reduced the DAI, CMDI, and HS scores (P<0.01 or P<0.05) of acute UC mice, reduced the level of IL-1ß and IL-17 in serum, increased the expression of IL-10 in serum and EGF in colon tissue, increased the number of CD3⁺ T cells, and decreased the level of CD4⁺ T cells and the ratio of CD4⁺/CD8⁺. CONCLUSIONS Kangfuxin liquid has a therapeutic effect on DSS-induced acute UC in mice, and its mechanism of action may be associated with regulating immune function and reducing intestinal inflammatory response.

Topics: Animals; Colitis, Ulcerative; Dextran Sulfate; Disease Models, Animal; Epidermal Growth Factor; Immunity; Inflammation; Interleukin-10; Interleukin-17; Materia Medica; Mice; Protective Agents; Signal Transduction

Topics: Cell Line; Epidermal Growth Factor; Epithelial Cells; Humans; Inflammation; Interleukin-6; Palmitic Acid; Sebaceous Glands; Signal Transduction

Partial burn injury in older patients is associated with higher rates of morbidity, mortality, and conversion to full thickness burn (Finnerty et al., 2009; Pham et al., 2009). Both human and mouse models demonstrate an altered systemic immune response in older subjects, however less is known about the localized response (Jeschke et al., 2016; Farinas et al., 2018; Mohs et al., 2017). We hypothesized that a mouse model could demonstrate differences in the localized inflammatory response of the old.. Six old (66 weeks) and young (8 weeks) mice received partial thickness thermal burns. Localized and systemic expression of nine chemokines (TNFalpha, MCP-1, MIP-2, S100A9, EGF, IL-10, RANTES, G-CSF, and EOTAXIN) were evaluated at day 3 after burn using Luminex analysis. Vimentin immunostaining was used to evaluate injury depth.. Vimentin staining demonstrated increased burn depth in old mice (449±38μm) as compared to young (166±18μm) (p<0.05). Both groups exhibited increased localized expression of EOTAXIN after burn (p<0.05), however expression in old mice (83.6±6.1pg/ml) was lower than that of young (126.8±18.7pg/ml) (p<0.05). Systemically, however, old mice had increased baseline EOTAXIN expression (1332.40±110.78pg/ml) compared to young (666.12±45.8pg/ml) (p<0.005).. EOTAXIN is one of the primary chemoattractants for selective eosinophilic recruitment and activation. While eosinophils are important for wound healing, a hyperactive eosinophilic response can result in tissue damage. We hypothesize that the increased baseline serum EOTAXIN in the old may prime their hyperactive response, and may contribute to their worse clinical outcomes. Long-term eosinophil activation requires further study, however our findings indicate a role for EOTAXIN and eosinophils in burn response.

Topics: Aging; Animals; Burns; Calgranulin B; Chemokine CCL11; Chemokine CCL2; Chemokine CCL24; Chemokine CCL26; Chemokine CCL5; Chemokine CXCL2; Eosinophils; Epidermal Growth Factor; Granulocyte Colony-Stimulating Factor; Inflammation; Interleukin-10; Mice; Tumor Necrosis Factor-alpha

To investigate the function and expression of the PGE2 receptors EP1-4 in rat retinal ischemia-reperfusion (I/R) injury and to determine the regulatory role of resveratrol (RES) in this process.. In vitro, we stimulated primary astrocytes extracted from the optic disc of rats with epidermal growth factor (EGF) and RES, and detected the location of EP1-4 expression with immunofluorescence. The expression of antiglial fibrillary acidic protein (GFAP), EGF receptor (EGFR), inducible NOS (iNOS), and EP1-4 in astrocytes was detected with western blotting. In vivo, we established an I/R injury model and RES treatment model with Sprague-Dawley rats. Changes in the thickness of the inner retina were observed with hematoxylin and eosin (H&E) staining. EP1-4 localization in the retina was observed with immunohistochemistry. The expression of COX-2, iNOS, and EP1-4 in the control and model groups was detected with western blotting.. In this study, immunofluorescence and immunohistochemistry showed that EP1-4 are expressed in astrocytes and the rat retina. EGF stimulation increased the expression of EGFR, iNOS, EP1, EP2, and EP4 in astrocytes. The expression of EP1-4 was statistically significantly increased on the third day after model induction, and EP1-4 expression decreased to normal levels on day 7. EGF and RES mediated the decrease in the expression of EP2. RES treatment significantly reduced retinal damage and RGC loss, as demonstrated by the relatively intact tissue structure on day 7 observed with H&E staining. Moreover, inflammation was associated with this I/R injury model, as demonstrated by the early induction of proinflammatory mediators, and this inflammation was significantly attenuated after RES treatment.. These results indicate that the COX-2/PGE2/EPs pathway is involved in retinal damage and astrocyte inflammation. In addition, the results suggest that the neuroprotective effects of RES may be associated with decreased production of inflammatory mediators. These results suggest that the PGE2 receptor may be a key factor in the treatment of neurodegenerative diseases, and that RES may be used as a possible therapeutic strategy for glaucoma.

Topics: Animals; Astrocytes; Cyclooxygenase 2; Disease Models, Animal; Epidermal Growth Factor; ErbB Receptors; Glial Fibrillary Acidic Protein; Humans; Immunohistochemistry; Inflammation; Male; Mice, Inbred C57BL; Nitric Oxide Synthase Type II; Optic Disk; Rats; Rats, Sprague-Dawley; Receptors, Prostaglandin E; Reperfusion Injury; Resveratrol; Retina; Signal Transduction

The purpose of this study is to reveal the participation of different regulatory cytokines within the process of pseudoexfoliation (PEX).. Our study included 140 patients referred to cataract surgery with early and late stage of pseudoexfoliation syndrome (XFS) or pseudoexfoliation glaucoma (XFG). Humor and serum levels of cytokines: transforming growth factor beta (TGF-β), platelet-derived growth factor (PDGF), epidermal growth factor (EGF), insulin-like growth factor (IGF), IL-8 and interferon-inducible T cell alpha chemoattractant (ITAC) were measured in a sample using high sensitivity enzyme-linked immunoabsorbent assay (ELISA) kit.. Our results indicate that profibrotic action induced by increasing TGF-β and PDGF locally activates fibrous tissue production in the early XFS with a prolonged effect of PDGF (late XFS) and finally (XFG stage) it is dominantly controlled by EGF and IGF. ITAC overrides angiogenetic effects of IL-8 in XFG.. Based on our findings, local chronic inflammation in the eye is accompanied by the secretion of different profibrotic cytokines (TGF-β, PDGF, EGF, IGF, IL-8) without angiogenesis due to effects of ITAC.

Topics: Cytokines; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Exfoliation Syndrome; Humans; Inflammation; Neovascularization, Physiologic; Platelet-Derived Growth Factor; Transforming Growth Factor beta

To investigate the effects of desmoglein 3 (DSG3) gene mediating epidermal growth factor/epidermal growth factor receptor (EGF/EGFR) signaling pathway on inflammatory response and immune function of anaphylactic rhinitis (AR).. Ten of the seventy male BALB/c mice were randomly selected as the normal control group, and the remaining 60 were used to construct the AR mice model. AR model mice were divided into 6 groups: model group (instilled with 5 μL saline), empty vector group (instilled with 5 μL of liposome and empty vector mixture), siRNA-DSG3 group (instilled with 5 μL of liposome and siRNA-DSG3 carrier mixture), AG1478 group (instilled with 5 μL of EGF/EGFR inhibitor AG1478), siRNA-DSG3+AG1478 group (instilled with 5 μL of liposome and siRNA-DSG3 carrier and EGF/EGFR inhibitor AG1478 mixture) and oe-DSG3 group, 10 in each group. After taking serum, each group of mice was sacrificed to get nasal mucosa tissues. HE staining was used to observe the pathological changes of nasal mucosa tissues in each group. The expression levels of DSG3, EGF and EGFR in nasal mucosa tissues of mice in each group were detected by qRT-PCR and western blot methods respectively. TUNEL staining was used to observe the apoptosis of nasal mucosa cells in mice. The expression of IgE, INF-γ, TNF-α, IL-2, IL-4 and IL-6 in serum of mice was determined by ELISA method. The immune adhesion function of red blood cells was detected by complement sensitization yeast hemagglutination method.. All the mice with AR showed different degrees of nasal mucosa injury and inflammatory cell infiltration, and silencing DSG3 or inhibiting the activity of EGF signaling pathway could alleviate the nasal mucosa injury. Compared with control group, the INF-γ and IL-2 levels of serum in AR model mice were significantly decreased; IgE, TNF-α, IL-4 and IL-6 levels were significantly increased (all P < 0.05); the mRNA expression levels and protein levels of DSG3, EGF and EGFR were significantly increased (all P < 0.05); C. Silencing DSG3 gene can inhibit the activation of EGF signaling pathway, alleviate the inflammation of AR nasal mucosa, and enhance red blood cells immune adherence function.

Topics: Anaphylaxis; Animals; Desmoglein 3; Disease Models, Animal; Epidermal Growth Factor; ErbB Receptors; Gene Expression Regulation; Inflammation; Mice, Inbred BALB C; Nasal Mucosa; Rhinitis, Allergic; Signal Transduction

In the present study, the effects of erlotinib on mouse tear function and corneal epithelial tissue structure were investigated. Throughout the 3 weeks of treatment, no notable differences were observed in the body, eye or lacrimal gland weights of the control and experimental mice. However, in the experimental group, the tear volume and break‑up times of tear film were significantly lower following treatment with erlotinib compared with the control group. Corneal fluorescein staining in the experimental group revealed patchy staining, and the Lissamine green staining and inflammatory index were significantly higher in the experimental group at 3 weeks than in the control group. In the experimental group, the number of corneal epithelium layers increased significantly following treatment with erlotinib for 3 weeks and a significant increase in the number of vacuoles was observed compared with the control group. Treatment with erlotinib significantly increased the corneal epithelial cell apoptosis, and led to a significantly increased number of epithelial cell layers and increased keratin 10 expression. It also significantly reduced the number of conjunctival goblet cells. Transmission electron microscopy and scanning electron microscopy revealed that the corneal epithelial surface was irregular and there was a substantial reduction and partial loss of the microvilli in the experimental group. Mice treated with erlotinib also exhibited an increased protein expression of tumor necrosis factor‑α and decreased protein expression of phosphorylated‑epidermal growth factor receptor in the corneal epithelial cells. The topical application of erlotinib eye drops was revealed to induce dry eyes in mice. This is a novel method of developing a model of dry eyes in mice.

Topics: Administration, Topical; Animals; Cell Count; Cornea; Disease Models, Animal; Dry Eye Syndromes; Epidermal Growth Factor; Epithelium; Erlotinib Hydrochloride; Goblet Cells; Inflammation; Male; Mice, Inbred BALB C; Ophthalmic Solutions; Phosphorylation; Tears; Tumor Necrosis Factor-alpha

To affect immune response and inflammation, the hepatitis C virus (HCV) substantially influences intercellular communication pathways that are decisive for immune cell recruitment. The present study investigates mechanisms by which HCV modulates chemokine-mediated intercellular communication from infected cells.. Chemokine expression was studied in HCV. HCV enhances expression of CXCR2 ligands in its host cell via the induction of epidermal growth factor (EGF) production. Knockdown of EGF or of the p65 subunit of the NF-κB complex results in a substantial downregulation of HCV-induced CXCR2 ligand expression, supporting the involvement of an EGF-dependent mechanism as well as activation of NF-κB. Furthermore, HCV upregulates expression of CXCR2 ligands in response to EGF stimulation via downregulation of the T-cell protein tyrosine phosphatase (TC-PTP [PTPN2]), activation of NF-κB, and enhancement of EGF-inducible signal transduction via MEK1 (MAP2K1). This results in the production of a cytokine/chemokine pattern by the HCV-infected cell that can recruit neutrophils but not monocytes.. These data reveal a novel EGF-dependent mechanism by which HCV influences chemokine-mediated intercellular communication. We propose that this mechanism contributes to modulation of the HCV-induced inflammation and the antiviral immune response.. In most cases hepatitis C virus (HCV) results in chronic infection and persistent viral replication, taking decades until development of overt disease. To achieve such a course, the respective virus must have developed mechanisms to circumvent antiviral response, to modulate the inflammatory response and to utilise the infrastructure of its host with moderate effect on its viability. The present study provides novel data indicating that HCV induces epidermal growth factor production in its host cell, enhancing epidermal growth factor-inducible expression of chemokines that bind to the CXCR2 receptor and recruit neutrophile granulocytes. Importantly, chemokines are critical mediators determining the pattern of immune cells recruited to the site of injury and thereby the local inflammatory and immunological milieu. These data strongly suggest that HCV triggers mechanisms that enable the virus to influence the inflammatory and immunological processes of its host.

Topics: Cell Communication; Cell Line; Epidermal Growth Factor; Hepacivirus; Hepatitis C, Chronic; Host Microbial Interactions; Humans; Immunity, Cellular; Inflammation; Receptors, Interleukin-8B; Signal Transduction; Up-Regulation; Virus Replication

Development of advanced wound dressing materials with rapid healing rates is in urgent demand for wound cares. A suitable microenvironment will promote cell proliferation and migration, which benefits to early wound healing and prevents inflammations and scars. In this work, N-carboxymethyl chitosan- and alginate-based hydrogels are prepared via both electrostatic interaction and divalent chelation with epidermal growth factor (EGF) payload to promote the cell proliferation and wound healing. The dual-crosslinked hydrogels are investigated in terms of rheology, water retention ability, and the release rate of EGF. Moreover, such amorphous hydrogel can promote cell proliferation and accelerate wound healing. The present study demonstrates that dual-crosslinked polysaccharide hydrogels are promising in wound care management.

Topics: Alginates; Animals; Bandages; Cell Movement; Cell Proliferation; Cellular Microenvironment; Chitosan; Cicatrix; Cross-Linking Reagents; Epidermal Growth Factor; Humans; Hydrogels; Inflammation; Mice; Polysaccharides; Wound Healing

Topics: Antiviral Agents; Epidermal Growth Factor; Hepacivirus; Hepatitis C; Humans; Inflammation; Ligands

Immune dysregulation plays a role in the vulnerability for mood disorders. Immune growth factors, such as Stem Cell Factor (SCF), Insulin-like Growth Factor-Binding Protein-2 (IGF-BP2), Epidermal Growth Factor (EGF), IL-7 and sCD25 have repeatedly been reported altered in patients with mood disorders. The aim of this study was to investigate levels of these factors in serum of adolescent bipolar offspring, who have a heightened risk for mood disorder development and to also analyze the data combined with previously published data. Growth factors were assessed by CBA/ELISA in adolescent bipolar offspring (n=96, mean age=16years) and in age- and gender-matched healthy controls (n=50). EGF belonged to a mutually correlating cluster of mainly neurotrophic compounds including S100B and BDNF, which were in general decreased in serum. IL-7, SCF, IGF-BP2 and sCD25, belonged to a different mutually correlating cluster of immune growth factors, which were in general increased: IGF-BP2 significantly in serum of offspring without a mood disorder, IL-7 and SCF in serum of offspring who had experienced a mood episode. This pattern of de- and increases was not different between bipolar offspring that developed or did not develop a mood disorder over time, apart from the IGF-BP2 level, which was near significantly higher in offspring later developing a mood disorder. Correlations with the previously published immune-cellular abnormalities were not found. In conclusion non-affected adolescents at familial mood disorder development risk were characterized by a distinct pattern of a series of compounds operating in a network of hematopoiesis, neurogenesis and inflammation.

Topics: Adolescent; Bipolar Disorder; Brain-Derived Neurotrophic Factor; Child of Impaired Parents; Epidermal Growth Factor; Female; Humans; Inflammation; Insulin-Like Growth Factor Binding Protein 2; Intercellular Signaling Peptides and Proteins; Interleukin-7; Male; S100 Calcium Binding Protein beta Subunit; Stem Cell Factor

To investigate the effect of debridement combined with recombinant human epidermal growth factor (rhEGF) on wound healing, scar formation, serum epidermal growth factor and inflammatory factors in patients with oral and maxillofacial trauma.. Eight-two cases with oral and maxillofacial trauma treated in our hospital (January 2015 to December 2016) were retrospectively analyzed. All patients received routine treatment, 41 received rhEGF besides routine treatment (experimental group), 41 patients received only cosmetic debridement and suturing (control group). The effect, wound healing time of 2 groups were recorded and compared. The serum epidermal growth factor, interleukin -6 (IL-6), tumor necrosis factor alpha (TNF-α) and interleukin -1 (Interleukin-1, IL-1) before and after operation were compared between 2 groups. SPSS16.0 software package was used to analyze the data.. The average wound healing time of the patients in the experimental group was 5.1±1.3 days, and the average wound healing time of patients in the control group was 7.4±1.9 days, the difference between 2 groups was statistically significant (t=6.397, P<0.01). The effective rate of the experimental group was significantly higher than the control group (P<0.05). Before operation, the serum epidermal growth factor, IL-6, TNF-α and IL-1 in 2 groups had no significant differences (P>0.05); 2 days and 5 days after operation, the serum epidermal growth factor level of the experimental group was significantly higher than the control group(P<0.05). The serum IL-6, TNF-α, IL-1 level of the experimental group were significantly lower than the control group(P<0.05).The side effects happened in the experimental group were significantly lower than control group (P<0.05).. Patients with oral and maxillofacial trauma treated by debridement combined with rhEGF can promote wound healing and reduce the degree of inflammation.

Topics: Cytokines; Debridement; Epidermal Growth Factor; Humans; Inflammation; Mouth; Recombinant Proteins; Retrospective Studies; Wound Healing

The pathogenesis of chronic rhinosinusitis (CRS) remains unclear to date. The tissue remodeling in nasal polyps may be the result of inflammatory mediators and may involve epithelial-mesenchymal transition (EMT) and EMT-associated features such as cell motility in nasal epithelial cells (NECs). We determined whether NEC in nasal polyps of CRS already display features of EMT in vivo or respond with EMT to growth factor stimulation in vitro. Nasal polyp tissues expressed both epithelial and mesenchymal markers. Primary NEC from inferior turbinates and nasal polyps responded to the EMT-inducing agents transforming growth factor (TGF)-β1 and epidermal growth factor (EGF) with different expression patterns of EMT markers (E-cadherin, N-cadherin, Snail, Slug, Twist), however, only NEC from nasal polyps were susceptible to TGF-β1 and EGF-dependent cell migration. Our data suggest that a partial EMT is associated with the pathogenesis of nasal polyps in CRS patients. Furthermore, we show for the first time that epithelial cells from both nasal polyps and inferior turbinates were able to undergo an EMT-like process following exposure to TGF-β1 or EGF in vitro but that only NEC from nasal polyps responded with enhanced cell motility. Our data suggest that NEC from CRS patients have undergo partial EMT and that this process may be involved in the pathogenesis of CRS.

Topics: Adult; Aged; Aged, 80 and over; Cell Movement; Epidermal Growth Factor; Epithelial Cells; Epithelial-Mesenchymal Transition; Female; Gene Expression Profiling; Humans; Inflammation; Male; Middle Aged; Nasal Polyps; Oligonucleotide Array Sequence Analysis; Sinusitis; Transforming Growth Factor beta1; Turbinates; Young Adult

There is a growing interest in low-grade inflammatory and metabolic alterations in patients with chronic schizophrenia (SCH).. Inflammatory (tumor-necrosis factor-α [TNF-α], interferon-γ [IFN-γ], interleukins [IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10], monocyte chemo-attractant protein-1 [MCP-1]) and growth factors (vascular endothelial growth factor [VEGF], epidermal growth factor [EGF]) were measured in blood serum samples of 105 SCH patients and 148 control subjects (CS). Simultaneously the clinical biomarkers (C-reactive protein [CRP], triglycerides [TG], low-density lipoprotein [LDL-c] and high-density lipoprotein [HDL-c] cholesterol, glycated hemoglobin [HbA1c]) were measured, and body mass index (BMI) was calculated for patients.. Several cyto-/chemokines (IFN-γ, MCP-1, IL-2, IL-6, IL-8 and IL-10) were significantly (P<0.0000001) elevated in SCH patients compared to CS. Odds ratios, obtained from logistic regression analyses, were significantly elevated for IL-2, IL-6, IL-10, INF-γ, and decreased for TNF-α in SCH group. Among the patients, higher IL-2, IL-6, INF-γ and lower MCP-1 levels as well as male gender were together significant (P<0.000001) predictors of higher HbA1c levels, and TG/HDL-c parameter was associated with ratios of INF-γ/IL-10 (P=0.004), and INF-γ/IL-4 (P=0.049), HbA1c (P=0.005), INF-γ (P=0.009), as well as LDL-c (P=0.02) levels.. IL-2, IL-6, IL-10 and IFN-γ were the most significant SCH-related markers among the measured cytokines in our patient group. Furthermore, significant associations between pro-/anti-inflammatory imbalance and HbA1c as well as cardio-metabolic risk marker (TG/HDL-c) were observed, indicating higher risks of diabetes and cardiovascular diseases among SCH patients.

Topics: Adult; Cytokines; Diabetes Mellitus, Type 2; Epidermal Growth Factor; Female; Humans; Inflammation; Male; Middle Aged; Obesity; Schizophrenia; Vascular Endothelial Growth Factor A

Thrombomodulin (TM) exerts anti-inflammatory functions. We previously found that recombinant human soluble TM alleviated murine graft-versus-host disease (GVHD). Nevertheless, it is unclear how TM mediates its anti-inflammatory functions in GVHD. Here, we identified G-protein coupled receptor 15 (GPR15) expressed on T cells as a receptor/sensor of TM. The fifth region of epidermal growth factor-like domain of TM (TME5) bound GPR15 in vitro. TME5 prolonged survival of mice undergoing acute GVHD after allogeneic hematopoietic stem cell transplantation (allo-HSCT). TME5 increased regulatory T cells (Tregs) but decreased Th 1 proportions in targeted organs. TME5 suppressed allo-reaction in vitro in association with an increase in the number of induced Tregs. However, the anti-inflammatory function of TME5 was abolished when GPR15 knockout T cells were used as donor T cells. We further found that TME5 suppressed production of IL-6 in T cells, which probably facilitated differentiation of Tregs. Moreover, TME5 reduced activation of bone marrow-derived dendritic cells (BMDCs) and hampered function of BMDCs in inducing allo-reaction in vivo and in vitro. Our findings suggested that inducing Tregs as well as blocking activation of DCs in vivo by using TME5 is a potential therapeutic option for preventing GVHD in allo-HSCT.

Topics: Animals; Dendritic Cells; Epidermal Growth Factor; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Humans; Inflammation; Leukocyte Reduction Procedures; Mice; Peptide Fragments; Receptors, G-Protein-Coupled; T-Lymphocytes, Regulatory; Thrombomodulin; Transplantation, Homologous

Epidermal growth factor (EGF)/EFG receptor (EGFR) signaling plays an important role in the osteoblastogenesis. The potential effects of pilose antler peptide (PAP) on osteoblast cell damages was investigated in our present study through EGF/EGFR signaling. In MC3T3-E1 osteoblastic cells, PAP treatment significantly inhibited the production of inflammatory cytokines by decreasing the levels of serum proinflammatory cytokines interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). PAP treatment also alleviated the oxidative responses as indicated by increased activities of catalase (SOD) and decreased levels of malondialdehyde (MDA). EGF inhibition, by siRNA knockdown, almost abolished PAP-induced osteoblast cytoprotection against inflammation and oxidant stress. Further, our results showed that PAP stimulated the nuclear erythroid factor 2-related factor 2 (Nrf2)2/heme oxygenase-1(HO-1) signaling, and inhibited the activation of uclear factor kappa B (NF-κB) pathway in MC3T3-E1 cells. On the other hand, EGF siRNA knockdown inhibited PAP-induced cytoprotection, which decreased the expression of Nrf-2, HO-1 and increased the level of p-NF-κBp65, p-IκBα in MC3T3-E1 cells. Thus, our research demonstrated that PAP protects osteoblasts from inflammatory and oxidative injury through EGF/EGFR signaling.

Topics: Animals; Antlers; Cell Line; Cell Survival; Cytokines; Cytoprotection; Deer; Epidermal Growth Factor; ErbB Receptors; Inflammation; Malondialdehyde; Mice; Osteoblasts; Oxidative Stress; Peptides; Signal Transduction; Superoxide Dismutase

Objective To observe the effects of Hedyotis diffusa extract (HDE) on the prolifera- tion, apoptosis, and inflammatory factors of HaCaT cells, and to explore its possible molecular mecha- nisms. Methods HaCaT cells were divided into 3 groups, the vehicle group, the control group, and 3 dose HDE groups. No epidermal growth factor (EGF) or HDE was added in the vehicle group. EGF was added with no HDE treatment in the control group. HDE (25, 50, 100 mg/mL) and EGF were added in the 3 HDE groups to co-culture HaCaT cells. The effects of HDE on EGF-induced proliferation of HaCaT cells were de- tected using CCK-8 assay. The effects of HDE on the growth cycle and apoptosis rate of HaCaT cells were measured using flow cytometry. Moreover, protein expression levels of Ki67, Bcl-xL, clAP1 , procaspase- 3, and cleaved caspase-3 were determined using Western blot. In addition, levels of IL-6, IL-8, and IL-10 in the supernatant were detected using ELISA. The level of phosphoration of NF-κB p65 (p-p65) was meas- ured using Western blot. Results Compared with the vehicle group, the absorbance of HaCaT cells and the expression level of Ki67 increased (P <0. 05, P <0. 01) ; levels of p-p65, IL-6, and IL-8 were elevated (P <0. 05, P <0. 01); IL-10 level was lowered (P <0.01) in the control group. Compared with the control group, the absorbance of HaCaT cells and the expression level of Ki67 decreased (P <0.05, P <0.01) ; levels of p-p65, IL-6, and IL-8 were reduced (P <0. 05, P <0. 01); IL-10 level was elevated (P <0. 05, P < 0.01) in the 3 dose HDE groups. Meanwhile, the apoptosis rate of HaCaT cells increased more in the 3 dose HDE groups than in the control group (P <0. 05, P <0. 01). The percentage of HaCaT cells at G1 phase was 58. 51 %, 73.12%, and 79. 95% in 25, 50, and 100 mg/mL HDE groups, respectively, showing statisti- cal difference when compared with that in the control group (52. 85%; P <0. 05, P <0. 01). The percentage and apoptosis rate of HaCaT cells at G1 phase were elevated more in 50 and 100 mg/mL HDE groups than in 25 mg/mL HDE group (P <0. 01). Besides, the percentage and apoptosis rate of HaCaT cells at G1 phase were elevated more in 100 mg/mL HDE group than in 50 mg/mL HDE group (P <0. 05). Compared with the control group, protein expression levels of Bcl-xL and cIAP1 were reduced in 100 mg/mL HDE group (P < 0. 01). There was no statistical difference in caspase-3 total amount (P >0. 05), but cleaved caspase-3 ex- pression increased with statistical

Topics: Apoptosis; Cell Proliferation; Cells, Cultured; Epidermal Growth Factor; Hedyotis; Inflammation; Plant Extracts; Tumor Necrosis Factor-alpha

Besides their importance for the vascular tone, vascular smooth muscle cells (VSMC) also contribute to pathophysiological vessel alterations. Various G-protein coupled receptor ligands involved in vascular dysfunction and remodeling can transactivate the epidermal growth factor receptor (EGFR) of VSMC, yet the importance of EGFR transactivation for the VSMC phenotype is incompletely understood. The aims of this study were (i) to characterize further the importance of the VSMC-EGFR for proliferation, migration and marker gene expression for inflammation, fibrosis and reactive oxygen species (ROS) homeostasis and (ii) to test the hypothesis that vasoactive substances (endothelin-1, phenylephrine, thrombin, vasopressin and ATP) rely differentially on the EGFR with respect to the abovementioned phenotypic alterations. In primary, aortic VSMC from mice without conditional deletion of the EGFR, proliferation, migration, marker gene expression (inflammation, fibrosis and ROS homeostasis) and cell signaling (ERK 1/2, intracellular calcium) were analyzed. VSMC-EGFR loss reduced collective cell migration and single cell migration probability, while no difference between the genotypes in single cell velocity, chemotaxis or marker gene expression could be observed under control conditions. EGF promoted proliferation, collective cell migration, chemokinesis and chemotaxis and leads to a proinflammatory gene expression profile in wildtype but not in knockout VSMC. Comparing the impact of five vasoactive substances (all reported to transactivate EGFR and all leading to an EGFR dependent increase in ERK1/2 phosphorylation), we demonstrate that the importance of EGFR for their action is substance-dependent and most apparent for crowd migration but plays a minor role for gene expression regulation.

Topics: Adenosine Triphosphate; Animals; Cell Movement; Cell Proliferation; Cells, Cultured; Endothelin-1; Enzyme Activation; Epidermal Growth Factor; ErbB Receptors; Extracellular Signal-Regulated MAP Kinases; Fibrosis; Gene Expression Regulation; Genotype; Inflammation; Ligands; Mice, Knockout; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Oxidative Stress; Phenotype; Phenylephrine; Primary Cell Culture; Signal Transduction; Thrombin; Time Factors; Vasopressins

The study monitored in vitro early response of connective tissue cells and immunocompetent cells to enosseal implant materials coated by different blood components (serum, activated plasma, and plasma/platelets) to evaluate human osteoblast proliferation and synthetic activity and inflammatory response presented as a cytokine profile of peripheral blood mononuclear cells (PBMCs) under conditions imitating the situation upon implantation. The cells were cultivated on coated Ti-plasma-sprayed (Ti-PS), Ti-etched (Ti-Etch), Ti-hydroxyapatite (Ti-HA), and ZrO2 surfaces. The plasma/platelets coating supported osteoblast proliferation only on osteoconductive Ti-HA and Ti-Etch whereas activated plasma enhanced proliferation on all surfaces. Differentiation (BAP) and IL-8 production remained unchanged or decreased irrespective of the coating and surface; only the serum and plasma/platelets-coated ZrO2 exhibited higher BAP and IL-8 expression. RANKL production increased on serum and activated plasma coatings. PBMCs produced especially cytokines playing role in inflammatory phase of wound healing, that is, IL-6, GRO-α, GRO, ENA-78, IL-8, GM-CSF, EGF, and MCP-1. Cytokine profiles were comparable for all tested surfaces; only ENA-78, IL-8, GM-CSF, and MCP-1 expression depended on materials and coatings. The activated plasma coating led to uniformed surfaces and represented a favorable treatment especially for bioinert Ti-PS and ZrO2 whereas all coatings had no distinctive effect on bioactive Ti-HA and Ti-Etch.

Topics: Cell Line; Cell Proliferation; Chemokine CCL2; Chemokine CXCL1; Chemokine CXCL5; Coated Materials, Biocompatible; Cytokines; Epidermal Growth Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation; Interleukin-6; Interleukin-8; Leukocytes, Mononuclear; Osteoblasts; Titanium; Wound Healing

The potential influence of leukocyte incorporation in the kinetic release of growth factors from platelet-rich plasma (PRP) may explain the conflicting efficiency of leukocyte platelet-rich plasma (L-PRP) scaffolds in tissue regeneration. To assess this hypothesis, leukocyte-free (PRGF-Endoret) and L-PRP fibrin scaffolds were prepared, and both morphogen and proinflammatory cytokine release kinetics were analyzed. Clots were incubated with culture medium to monitor protein release over 8 days. Furthermore, the different fibrin scaffolds were morphologically characterized. Results show that leukocyte-free fibrin matrices were homogenous while leukocyte-containing ones were heterogeneous, loose and cellular. Leukocyte incorporation produced a significant increase in the contents of proinflammatory cytokines interleukin (IL)-1β and IL-16 but not in the platelet-derived growth factors release (<1.5-fold). Surprisingly, the availability of vascular endothelial growth factor suffered an important decrease after 3 days of incubation in the case of L-PRP matrices. While the release of proinflammatory cytokines was almost absent or very low from PRGF-Endoret, the inclusion of leukocytes induced a major increase in these cytokines, which was characterized by the presence of a latent period. The PRGF-Endoret matrices were stable during the 8 days of incubation. The inclusion of leukocytes alters the growth factors release profile and also increased the dose of proinflammatory cytokines.

Topics: Cell Adhesion; Cytokines; Epidermal Growth Factor; Fibrin; Hepatocyte Growth Factor; Humans; Hydrogels; Inflammation; Insulin; Insulin-Like Growth Factor I; Intercellular Signaling Peptides and Proteins; Interleukin-16; Interleukin-1beta; Leukocytes; Optics and Photonics; Platelet-Derived Growth Factor; Platelet-Rich Plasma; Tissue Engineering; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A

Non-alcoholic fatty liver disease (NAFLD) defines a wide spectrum of liver diseases that extends from simple steatosis to non-alcoholic steatohepatitis. Although the pathogenesis of NAFLD remains undefined, it is recognized that insulin resistance is present in almost all patients who develop this disease. Thiazolidinediones (TZDs) act as an insulin sensitizer and have been used in the treatment of patients with type 2 diabetes and other insulin-resistant conditions, including NAFLD. Hence, therapy of NAFLD with insulin-sensitizing drugs should ideally improve the key hepatic histological changes, while also reducing cardiometabolic and cancer risks. Controversially, TZDs are associated with the development of cardiovascular events and liver problems. Therefore, there is a need for the development of new therapeutic strategies to improve liver function in patients with chronic liver diseases. The aim of the present study was to assess the therapeutic effects of LPSF/GQ-02 on the liver of LDLR-/- mice after a high-fat diet. Eighty male mice were divided into 4 groups and two different experiments: 1-received a standard diet; 2-fed with a high-fat diet (HFD); 3-HFD+pioglitazone; 4-HFD+LPSF/GQ-02. The experiments were conducted for 10 or 12 weeks and in the last two or four weeks respectively, the drugs were administered daily by gavage. The results obtained with an NAFLD murine model indicated that LPSF/GQ-02 was effective in improving the hepatic architecture, decreasing fat accumulation, reducing the amount of collagen, decreasing inflammation by reducing IL-6, iNOS, COX-2 and F4 / 80, and increasing the protein expression of IκBα, cytoplasmic NFκB-65, eNOS and IRS-1 in mice LDLR -/-. These results suggest a direct action by LPSF/GQ-02 on the factors that affect inflammation, insulin resistance and fat accumulation in the liver of these animals. Further studies are being conducted in our laboratory to investigate the possible mechanism of action of LPSF/GQ-02 on hepatic lipid metabolism.

Topics: Animals; Cyclooxygenase 2; Diet, High-Fat; Disease Models, Animal; Epidermal Growth Factor; I-kappa B Proteins; Inflammation; Interleukin-6; Liver; Male; Mice; Mice, Inbred C57BL; Mice, Knockout; NF-KappaB Inhibitor alpha; Nitric Oxide Synthase Type II; Non-alcoholic Fatty Liver Disease; Pioglitazone; Receptors, LDL; Thiazolidinediones; Triglycerides

We examined the effect of epidermal growth factor (EGF) and bone marrow transplantation (BMT) on gastrointestinal damage after high-dose irradiation of mice.. C57Black/6 mice were used. Two survival experiments were performed (12 and 13 Gy; (60)Co, 0.59-0.57 Gy/min). To evaluate BMT and EGF action, five groups were established - 0 Gy, 13 Gy, 13 Gy + EGF (at 2 mg/kg, first dose 24 h after irradiation and then every 48 h), 13 Gy + BMT (5 × 10(6) cells from green fluorescent protein [GFP] syngenic mice, 4 h after irradiation), and 13 Gy + BMT + EGF. Survival data, blood cell counts, gastrointestine and liver parameters and GFP positive cell migration were measured.. BMT and EGF (three doses, at 2 mg/kg, administered 1, 3 and 5 days after irradiation) significantly increased survival (13 Gy). In blood, progressive cytopenia was observed with BMT, EGF or their combination having no improving effect early after irradiation. In gastrointestinal system, BMT, EGF and their combination attenuated radiation-induced atrophy and increased regeneration during first week after irradiation with the combination being most effective. Signs of systemic inflammatory reaction were observed 30 days after irradiation.. Our data indicate that BMT together with EGF is a promising strategy in the treatment of high-dose whole-body irradiation damage.

Topics: Animals; Apoptosis; Bone Marrow Transplantation; Combined Modality Therapy; Epidermal Growth Factor; Female; Gastrointestinal Tract; Inflammation; Lithostathine; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mitosis; Radiation Injuries, Experimental; Whole-Body Irradiation

Amphiregulin (Areg) participates in tissue repair and inflammation regulation. As important effector cells in inflammation, macrophages can be polarized to classically (M1) or alternatively (M2) activated phenotype with diverse functions in immunity. However, the relationship between Areg expression and macrophage activation is poorly understood. Here we report that Areg was significantly expressed in M1 but not in M2 macrophages. This was confirmed by analyses of RT-PCR and ELISA in peritoneal macrophages, and by evaluating protein expression in alveolar macrophages and RAW264.7 cells. Selective inhibitors of TLR4 (CLI-095) and MAP kinase, including Erk1/2 (PD98059), JNK (SP600125) and p38 (SB203580), significantly reduced Areg expression in M1 macrophages, suggesting that M1 macrophages produce Areg mainly through the TLR4-MAPK pathway, which is involved in the mechanism of M1 activation. When compared with productions of classical biomarkers of M1 macrophages, Areg expression was highly consistent in time series. Taken together, Areg may be an effective new biomarker of M1 macrophages.

Topics: Amphiregulin; Animals; Biomarkers; Cell Differentiation; Cell Line; Cytokines; Epidermal Growth Factor; Gene Expression Regulation; Inflammation; Macrophage Activation; Macrophages; Macrophages, Alveolar; Macrophages, Peritoneal; Male; Mice; Mice, Inbred C57BL; Phenotype; Signal Transduction; Th2 Cells; Time Factors; Toll-Like Receptor 4

Epidermal growth factor (EGF) and their receptor (EGFR) play an important role in the development of cancer proliferation, and metastasis, although the mechanism remains unclear. The present study aimed at investigating the role of EGF-EGFR signalling pathway in the development of human hepatocellular carcinoma (HCC) inflammatory environment. Gene profiles of inflammatory cytokines from HCC were measured. Cell bio-behaviours of HCC with low or high metastasis were detected by the live cell monitoring system. Cell proliferation was measured by CCK8. The protein level of CXCL5 and CXCL8 was measured by ELISA. The phosphorylation of PI3K, ERK, MAPK was measured by western blot. EGF significantly induced cell proliferation in HepG2 cells, but not in HCCLM3 cells. EGF prompted the cell movement in both HepG2 and HCCLM3 and regulated the production of CXCL5 and CXCL8 from HCC, which were inhibited by EGFR inhibitor, Erk inhibitor (U0126), or PI3K inhibitors (BEZ-235 and SHBM1009). HCC proliferation, metastasis and production of inflammatory cytokines were regulated via EGF-EGFR signal pathways. CXCL5 could interact with CXCL8, possibly by CXCR2 or the cross-talk between CXCR2 and EGFR. EGF-EGFR signaling pathway can be the potential target of therapies for HCC.

Topics: Butadienes; Carcinoma, Hepatocellular; Cell Line, Tumor; Cell Movement; Cell Proliferation; Chemokine CXCL5; Enzyme Inhibitors; Epidermal Growth Factor; ErbB Receptors; Extracellular Signal-Regulated MAP Kinases; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Imidazoles; Inflammation; Interleukin-8; Liver Neoplasms; Nitriles; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Phosphorylation; Quinolines; Receptor Cross-Talk; Receptors, Interleukin-8B; Signal Transduction; Tumor Microenvironment

Monocyte/macrophage accumulation plays a critical role during progression of cardiovascular diseases, such as atherosclerosis. Our previous studies demonstrated that retrovirally mediated expression of the versican V3 splice variant (V3) by arterial smooth muscle cells (ASMCs) decreases monocyte adhesion in vitro and macrophage accumulation in a model of lipid-induced neointimal formation in vivo. We now demonstrate that V3-expressing ASMCs resist monocyte adhesion by altering the composition of the microenvironment surrounding the cells by affecting multiple signaling pathways. Reduction of monocyte adhesion to V3-expressing ASMCs is due to the generation of an extracellular matrix enriched in elastic fibers and depleted in hyaluronan, and reduction of the proinflammatory cell surface vascular cell adhesion molecule 1 (VCAM1). Blocking these changes reverses the protective effect of V3 on monocyte adhesion. The enhanced elastogenesis induced by V3 expression is mediated by TGFβ signaling, whereas the reduction in hyaluronan cable formation induced by V3 expression is mediated by the blockade of epidermal growth factor receptor and NFκB activation pathways. In addition, expression of V3 by ASMCs induced a marked decrease in NFκB-responsive proinflammatory cell surface molecules that mediate monocyte adhesion, such as VCAM1. Overall, these results indicate that V3 expression by ASMCs creates a microenvironment resistant to monocyte adhesion via differentially regulating multiple signaling pathways.

Topics: Animals; Cell Adhesion; Cells, Cultured; Cellular Microenvironment; Epidermal Growth Factor; Inflammation; Monocytes; Muscle, Smooth, Vascular; NF-kappa B; Rats; Rats, Inbred F344; Signal Transduction; Transforming Growth Factor beta1; Versicans

Growth factors are important for regulating a variety of cellular processes and typically act as signaling molecules between cells. In the present study, we examined the mechanisms underlying the inhibitory effects of mixtures of recombinant growth factors (MRGFs) on nitric oxide (NO) and pro-inflammatory cytokine production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. We also examined whether these effects are mediated through the mitogen‑activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) signal transduction pathways. NO production was assessed by measuring nitrite acucmulation using the Greiss reaction. Cytokine concentrations were measured using respective ELISA kits for each cytokine. Our results revealed that the MRGFs significantly attenuated the LPS-induced production of pro-inflammatory cytokines and NO in a dose-dependent manner. To elucidate the mechanisms underlying the inhibitory effects of MRGFs, we examined the effects of the LPS-induced phosphorylation of MAPKs and the activation of the NF-κB signaling pathway on the stabilization of NF-κB nuclear translocation and inhibitory factor-κB (IκB) degradation. Western blot analysis was performed to determine the total and phosphorylated levels of ERK, as well as the nuclear translocation of NF-κB, and IκB phosphorylation and degradation. Our results demonstrated that treatment with MRGFs resulted in a reduction in the phosphorylation of the ERK and NF-κB signaling pathways, whereas the phosphorylation of JNK and p38 was not affected. Taken together, our results suggest that MRGFs inhibit the production of pro-inflammatory cytokines and NO by downregulating inducible NO synthase gene expression and blocking the phosphorylation of the ERK and NF-κB signaling pathways. These findings may provide direct evidence of the potential application of MRGFs in the prevention and treatment of inflammatory diseases.

Topics: Animals; Cytokines; Epidermal Growth Factor; Inflammation; Insulin; Insulin-Like Growth Factor I; Lipopolysaccharides; Metabolism, Inborn Errors; Mice; Mitogen-Activated Protein Kinases; NF-kappa B; Nitric Oxide; Nitric Oxide Synthase Type II; Signal Transduction

Otitis media is one of the most common pediatric infections. While it is usually treated without difficulty, up to 20% of children may progress to long-term complications that include hearing loss, impaired speech and language development, academic underachievement, and irreversible disease. Hyperplasia of middle ear mucosa contributes to the sequelae of acute otitis media and is of important clinical significance. Understanding the role of growth factors in the mediation of mucosal hyperplasia could lead to the development of new therapeutic interventions for this disease and its sequelae.. From a whole genome gene array analysis of mRNA expression during acute otitis media, we identified growth factors with expression kinetics temporally related to hyperplasia. We then tested these factors for their ability to stimulate mucosal epithelial growth in vitro, and determined protein levels and histological distribution in vivo for active factors.. From the gene array, we identified seven candidate growth factors with upregulation of mRNA expression kinetics related to mucosal hyperplasia. Of the seven, only HB-EGF (heparin-binding-epidermal growth factor) induced significant mucosal epithelial hyperplasia in vitro. Subsequent quantification of HB-EGF protein expression in vivo via Western blot analysis confirmed that the protein is highly expressed from 6 hours to 24 hours after bacterial inoculation, while immunohistochemistry revealed production by middle ear epithelial cells and infiltrating lymphocytes.. Our data suggest an active role for HB-EGF in the hyperplasia of the middle ear mucosal epithelium during otitis media. These results imply that therapies targeting HB-EGF could ameliorate mucosal growth during otitis media, and thereby reduce detrimental sequelae of this childhood disease.

Topics: Animals; Disease Models, Animal; Ear, Middle; Epidermal Growth Factor; Epithelium; Heparin-binding EGF-like Growth Factor; Hyperplasia; Inflammation; Mice; Mice, Inbred C57BL; Mucous Membrane; Otitis Media; Rats, Sprague-Dawley; RNA, Messenger

The purpose of this study was to investigate the effects of an epidermal growth factor (EGF) intervention on improving the inflammatory response of rats fed an ethanol-containing diet. Eight-week-old male Wistar rats were divided into ethanol (E) and control (C) groups. Rats in the E group were fed an ethanol liquid diet, while rats in the C group were pair-fed an isoenergetic diet without ethanol. After a 4-week ethanol-induction period, both the C and E group were respectively subdivided into 2 groups: a normal liquid diet without (C group, n = 8) or with EGF supplementation (C + EGF, n = 8), and the ethanol-containing diet without (E group, n = 8) or with EGF supplementation (E + EGF group, n = 8). The EGF (30 μg/kg body weight/day) intervention period was carried out for the following 8 weeks. At the end of the experiment, activity of aspartate transaminase (AST) and alanine transaminase (ALT) and hepatic levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-10 in group E were significantly higher than those in group C. In addition, alterations in the gut microbiota profile were found in group E. In contrast, activity of AST and ALT and levels of TNF-α, IL-1β, and IL-6 in group E + EGF were significantly lower than those in group E. Significantly lower intestinal permeability and lower numbers of Escherichia coli in the fecal microbial culture were also found in group E + EGF. These results suggest that EGF improved the intestinal integrity by decreasing E. coli colonization and lowering intestinal permeability, which then ameliorated the inflammatory response under chronic ethanol exposure.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Epidermal Growth Factor; Ethanol; Inflammation; Interleukin-10; Interleukin-1beta; Interleukin-6; Intestines; Liver; Male; Microbiota; Rats, Wistar; Tumor Necrosis Factor-alpha

Hypertension is an important risk factor for cardiovascular disease and there is increasing evidence that inflammation and abnormal immune responses are involved in the pathogenesis of hypertension. However, the data on the association between specific cytokine concentrations and hypertension are inconsistent. We have evaluated the association between 12 cytokines/growth factors and the presence of different degrees of hypertension, comparing these concentrations to values in a healthy group of subjects. The concentrations of interleukin (IL)-1α, -1β, -2, -4, -6, -8, -10, tumor necrosis factor (TNF-α), interferon-γ (IFN-γ), monocyte chemoattractant protein (MCP-1), epidermal growth factor, and vascular endothelial growth factor were measured in 155 hypertensive patients and 148 healthy subjects, using EV-3513 cytokine biochip arrays, a competitive chemiluminescence immunoassay. Univariate and multivariate analyses were used to evaluate the association of specific cytokines and growth factors with systolic blood pressure (SBP) and diastolic blood pressure (DBP). Hypertensive subjects had higher serum concentrations of IL-1α, -2, -8, vascular endothelial growth factor, IFN-γ, TNF-α, MCP-1, and epidermal growth factor; and lower concentrations of anti-inflammatory cytokine, IL-10 (P < .05), compared with the healthy individuals. The serum concentrations of IL-4, -6, and -1β did not differ between the hypertensive subjects and control group. Univariate and multivariate analyses revealed that IL-1α and IFN-γ were independent predictors of a high SBP, while IFN-γ, IL-1α, TNF-α, and MCP-1 remained statistically significant for DBP after correction for age, gender, Body mass index, smoking, fasting blood glucose, and triglycerides. There was a significant association between the concentrations of several cytokines and hypertension. These associations may either be related to common underlying factors that cause hypertension and may also be proinflammatory or because these inflammatory cytokines might directly be involved in the etiology of hypertension.

Topics: Biomarkers; Blood Pressure; C-Reactive Protein; Cytokines; Epidermal Growth Factor; Female; Humans; Hypertension; Immunoassay; Inflammation; Male; Middle Aged; Prognosis; Vascular Endothelial Growth Factor A

Histone deacetylases (HDACs) regulate gene transcription by modifying the acetylation of histone and nonhistone proteins. Deregulated expression of HDACs has been implicated in tumorigenesis and angiogenesis. In this study, we examined the effect of suberoylanilide hydroxamic acid (SAHA), a potent inhibitor of HDACs, on inflammatory corneal angiogenesis. In a mouse model of alkali-induced corneal neovascularization (CNV), topical application of SAHA to the injured corneas attenuated CNV. In addition, in vivo treatment with SAHA downregulated the expression of the pro-angiogenic factors vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), transforming growth factor beta 1 (TGFβ1), and epidermal growth factor (EGF), but upregulated the expression of the anti-angiogenic factors thrombospondin (TSP)-1, TSP-2, and ADAMTS-1 in the injured corneas. Furthermore, SAHA inhibited the expression of pro-angiogenic factors, migration, proliferation, and tube formation by human microvascular endothelial cells (HEMC-1) in vitro. These data indicate that SAHA has therapeutic potential for CNV.

Topics: ADAM Proteins; ADAMTS1 Protein; Administration, Ophthalmic; Animals; Apoptosis; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; Cell Movement; Cell Proliferation; Cornea; Epidermal Growth Factor; Female; Histone Deacetylases; Hydroxamic Acids; Inflammation; Mice, Inbred BALB C; Neovascularization, Pathologic; Thrombospondins; Transforming Growth Factor beta1; Vascular Endothelial Growth Factor A; Vorinostat

The IL-6-triggered positive feedback loop for NFκB signaling (or the IL-6 amplifier/Inflammation amplifier) was originally discovered as a synergistic-activation signal that follows IL-17/IL-6 stimulation in nonimmune cells. Subsequent results from animal models have shown that the amplifier is activated by stimulation of NFκB and STAT3 and induces chemokines and inflammation via an NFκB loop. However, its role in human diseases is unclear. Here, we combined two genome-wide mouse screens with SNP-based disease association studies, revealing 1,700 genes related to the IL-6 amplifier, 202 of which showed 492 indications of association with ailments beyond autoimmune diseases. We followed up on ErbB1 from our list. Blocking ErbB1 signaling suppressed the IL-6 amplifier, whereas the expression of epiregulin, an ErbB1 ligand, was higher in patients with inflammatory diseases. These results indicate that the IL-6 amplifier is indeed associated with human diseases and disorders and that the identified genes may make for potential therapeutic targets.

Topics: Animals; Arthritis, Rheumatoid; Cell Line; Epidermal Growth Factor; Epiregulin; Epistasis, Genetic; Feedback, Physiological; Genes, erbB-1; Genetic Association Studies; Genetic Loci; Genome; Humans; Inflammation; Interleukin-6; Mice; Mice, Inbred C57BL; Multiple Sclerosis; Polymorphism, Single Nucleotide; Signal Transduction; Transcription, Genetic

Previous epidemiological studies together with animal studies have suggested an association between adiponectin and oxidative stress and inflammation, but community-based studies are lacking. Our objective was to investigate the relative importance of oxidative stress and inflammatory markers, representing different pathways in relation to adiponectin.. In a cross-sectional sample of 929 70-year-old individuals (50% women) of the Prospective Investigation of the Vasculature in Uppsala Seniors study, relations between serum adiponectin and oxidative stress [conjugated dienes (CD), homocysteine, total antioxidant capacity, oxidized low-density lipoprotein (OxLDL), OxLDL antibodies, baseline CD of LDL, glutathione (GSH), total glutathione (TGSH), glutathione disulfide], circulation interleukins (IL-6, IL-8), other cytokines [tumor necrosis factor α, monocyte chemotactic protein-1 (MCP-1), epidermal growth factor (EGF), vascular endothelial growth factor], cell adhesion molecules (vascular cell adhesion molecule-1, intercellular adhesion molecule-1, E-selectin, P-selectin, L-selectin), and systemic inflammatory markers [C-reactive protein (CRP), leukocyte count] in separate models were investigated.. In age- and sex-adjusted, as well as multivariable-adjusted models, adiponectin was significantly and positively associated with GSH, log TGSH, whereas an inverse association was observed for CD and log EGF. An inverse association between adiponectin and MCP-1, log E-selectin, and log CRP was significant in age- and sex-adjusted models, but not in multivariable-adjusted models.. Our results imply that higher levels of adiponectin are associated with a more beneficial oxidative stress profile, with higher levels of principal anti-oxidative GSH and total GSH together with lower levels of lipid peroxidation, possibly through shared pathways. Further studies are needed to investigate whether changes in the oxidative stress profile may be a mechanism linking adiponectin with type 2 diabetes and/or cardiovascular disease.

Topics: Adiponectin; Aged; Biomarkers; C-Reactive Protein; Cell Adhesion Molecules; Chemokine CCL2; Cross-Sectional Studies; E-Selectin; Epidermal Growth Factor; Female; Glutathione; Humans; Inflammation; Intercellular Adhesion Molecule-1; Interleukin-6; Interleukin-8; L-Selectin; Linear Models; Lipoproteins, LDL; Male; Multivariate Analysis; Oxidative Stress; P-Selectin; Prospective Studies; Tumor Necrosis Factor-alpha; Vascular Cell Adhesion Molecule-1

Interactions between dendritic cells (DCs) and T cells play a critical role in the development of glomerulonephritis, which is a common cause of chronic kidney disease. DC-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN), an immune-regulating molecule of the C-type lectin family, is mainly expressed on DCs and mediates DC adhesion and migration, inflammation, activation of primary T cells. DC-SIGN triggers immune responses and is involved in the immune escape of pathogens and tumours. In addition, ligation of DC-SIGN on DCs actively primes DCs to induce Tregs. Under certain conditions, DC-SIGN signalling may result in inhibition of DC maturation, by promoting regulatory T cell (Treg) function and affecting Th1/Th2 bias.. A rat model of nephrotoxic nephritis was used to investigate the therapeutic effects of an anti-lectin-epidermal growth factor (EGF) antibody on glomerulonephritis. DCs were induced by human peripheral blood mononuclear cells in vitro. The expression of DC surface antigens were detected using flow cytometry; the levels of cytokines were detected by ELISA and qPCR, respectively; the capability of DCs to stimulate T cell proliferation was examined by mixed lymphocyte reaction; PsL-EGFmAb targeting to DC-SIGN on DCs was identified by immunoprecipitation.. Anti-Lectin-EGF antibody significantly reduced global crescent formation, tubulointerstitial injury and improved renal function impairment through inhibiting DC maturation and modulating Foxp3 expression and the Th1/Th2 cytokine balance in kidney. Binding of anti-Lectin-EGF antibody to DC-SIGN on human DCs inhibited DC maturation, increased IL-10 production from DCs and enhanced CD4+CD25+ Treg functions.. Our results suggest that treatment with anti-Lectin-EGF antibody modulates DCs to suppressive DCs and enhances Treg functions, contributing to the attenuation of renal injury in a rat model of nephrotoxic nephritis.

Topics: Animals; Antibodies; Antibodies, Monoclonal; Antigens, Surface; CD4-Positive T-Lymphocytes; Cell Adhesion Molecules; Dendritic Cells; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Humans; Inflammation; Interleukin-2 Receptor alpha Subunit; Kidney; Lectins, C-Type; Male; Nephritis; Protein Structure, Tertiary; Rats; Rats, Inbred WKY; Receptors, Cell Surface; Renal Insufficiency, Chronic; Signal Transduction; T-Lymphocytes, Regulatory; Th1 Cells; Th2 Cells

Selenium (Se) is an essential micronutrient that exerts its functions via selenoproteins. Little is known about the role of Se in inflammatory bowel disease (IBD). Epidemiological studies have inversely correlated nutritional Se status with IBD severity and colon cancer risk. Moreover, molecular studies have revealed that Se deficiency activates WNT signaling, a pathway essential to intestinal stem cell programs and pivotal to injury recovery processes in IBD that is also activated in inflammatory neoplastic transformation. In order to better understand the role of Se in epithelial injury and tumorigenesis resulting from inflammatory stimuli, we examined colonic phenotypes in Se-deficient or -sufficient mice in response to dextran sodium sulfate (DSS)-induced colitis, and azoxymethane (AOM) followed by cyclical administration of DSS, respectively. In response to DSS alone, Se-deficient mice demonstrated increased morbidity, weight loss, stool scores, and colonic injury with a concomitant increase in DNA damage and increases in inflammation-related cytokines. As there was an increase in DNA damage as well as expression of several EGF and TGF-β pathway genes in response to inflammatory injury, we sought to determine if tumorigenesis was altered in the setting of inflammatory carcinogenesis. Se-deficient mice subjected to AOM/DSS treatment to model colitis-associated cancer (CAC) had increased tumor number, though not size, as well as increased incidence of high grade dysplasia. This increase in tumor initiation was likely due to a general increase in colonic DNA damage, as increased 8-OHdG staining was seen in Se-deficient tumors and adjacent, non-tumor mucosa. Taken together, our results indicate that Se deficiency worsens experimental colitis and promotes tumor development and progression in inflammatory carcinogenesis.

Topics: 8-Hydroxy-2'-Deoxyguanosine; Animals; Azoxymethane; Carcinogenesis; Colitis; Colonic Neoplasms; Deoxyguanosine; Dextran Sulfate; Diet; DNA Damage; Epidermal Growth Factor; Gene Expression Regulation; Inflammation; Mice; Mice, Inbred C57BL; Selenium; Signal Transduction; Transforming Growth Factor beta; Weight Loss

Breast cancer progression is strongly linked to inflammatory processes, aggravating disease course. The impacts of the inflammatory cytokine TNF α on breast malignancy are not fully substantiated, and they may be affected by cooperativity between TNF α and other protumoral mediators. Here, we show that together with representatives of other important arms of the tumor microenvironment, estrogen (hormonal) and EGF (growth-supporting), TNF α potently induced metastasis-related properties and functions in luminal breast tumor cells, representing the most common type of breast cancer. Jointly, TNFα + Estrogen + EGF had a stronger effect on breast cancer cells than each element alone, leading to the following: (1) extensive cell spreading and formation of FAK/paxillin-enriched cellular protrusions; (2) elevated proportion of tumor cells coexpressing high levels of CD44 and β 1 and VLA6; (3) EMT and cell migration; (4) resistance to chemotherapy; (5) release of protumoral factors (CXCL8, CCL2, MMPs). Importantly, the tumor cells used in this study are known to be nonmetastatic under all conditions; nevertheless, they have acquired high metastasizing abilities in vivo in mice, following a brief stimulation by TNFα + Estrogen + EGF. These dramatic findings indicate that TNF α can turn into a strong prometastatic factor, suggesting a paradigm shift in which clinically approved inhibitors of TNFα would be applied in breast cancer therapy.

Topics: Animals; Antineoplastic Agents; Breast Neoplasms; Cell Survival; Disease Progression; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Estrogens; Female; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Inflammation; Integrin beta1; MCF-7 Cells; Mice; Mice, Nude; Microscopy, Confocal; Neoplasm Metastasis; Neoplasm Transplantation; Spheroids, Cellular; Tumor Microenvironment; Tumor Necrosis Factor-alpha

This study determined whether N-acetylcysteine (NAC) could affect intestinal redox status, proinflammatory cytokines, epidermal growth factor (EGF), EGF receptor (EGFR), Toll-like receptor-4 (TLR4), and aquaporin-8 in a lipopolysaccharide (LPS)-challenged piglet model. Eighteen piglets (35-day-old) were randomly allocated into one of the three treatments (control, LPS and NAC). The control and LPS groups were fed a basal diet, and the NAC group received the basal diet +500 mg/kg NAC. On days 10, 13, and 20 of the trial, the LPS- and NAC-treated piglets received intraperitoneal administration of LPS (100 μg/kg BW), whereas the control group received the same volume of saline. On days 10 and 20, venous blood samples were obtained at 3 h post LPS or saline injection. On day 21 of the trial, piglets were killed to obtain the intestinal mucosa for analysis. Compared with the control group, LPS challenge reduced (P < 0.05) the activities of superoxide dismutase, catalase, and glutathione peroxidase in jejunal mucosae, while increasing (P < 0.05) the concentrations of malondialdehyde, H2O2, O2 (·-) and the ratio of oxidized to reduced glutathione in jejunal mucosae, and concentrations of TNF-α, cortisol, interleukin-6, and prostaglandin E2 in both plasma and intestinal mucosae. These adverse effects of LPS were attenuated (P < 0.05) by NAC supplementation. Moreover, NAC prevented LPS-induced increases in abundances of intestinal HSP70 and NF-κB p65 proteins and TLR4 mRNA. NAC supplementation enhanced plasma EGF concentration and intestinal EGFR mRNA levels. Collectively, these results indicate that dietary NAC supplementation alleviates LPS-induced intestinal inflammation via regulating redox, EGF, and TLR4 signaling.

Topics: Acetylcysteine; Animals; Dietary Supplements; Epidermal Growth Factor; Female; Inflammation; Intestine, Small; Lipopolysaccharides; Oxidation-Reduction; Real-Time Polymerase Chain Reaction; RNA, Messenger; Signal Transduction; Swine; Toll-Like Receptor 4

Epidermal growth factor (EGF) is an attractive and promising therapeutic application for intestinal disorders. The current study examined its influence on proliferation and restoration after ischemia-reperfusion (I/R) injury in rat small intestine. Six groups were performed: sham operation (Con); ischemia for 30 min with subsequent reperfusion for 30 min (I/R); I/R injured with 500 μg/kg EGF injected 5 min before ischemia (Pre-l); I/R injured with 50 μg/kg EGF injected 5 min before ischemia (Pre-s); I/R injured with 500 μg/kg EGF injected 5 min after reperfusion (Post-l); and I/R injured with 50 μg/kg EGF injected 5 min after reperfusion (Post-s). Intestinal histological damage, crypt cell proliferation degree, mucosal permeability, tight junction proteins expression, and levels of inflammation factors were studied for each group. Compared with the I/R group, administration of EGF in the Pre-l, Pre-s, and Post-l groups all presented a significant proliferation effect. The levels of FD4, IL-6, and TNF-α were dramatically decreased in all EGF-treated groups. Histological destruction was improved and TJs recovery was notably accelerated in all EGF-treated groups except the Post-s group. D-lactate concentration was only diminished in the Pre-l group. These results suggest that mucosally applied EGF can promote intestinal proliferation and improve restoration after I/R injury. EGF intraluminal administration is an effective treatment against intestinal I/R injury.

Topics: Animals; Cell Proliferation; Epidermal Growth Factor; Inflammation; Interleukin-6; Intestinal Mucosa; Intestines; Male; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Tight Junction Proteins; Tight Junctions; Tumor Necrosis Factor-alpha

Extracts from Serenoa repens are widely used for the treatment of benign prostatic hyperplasia (BPH) and traditionally for prostatitis. In the present study we evaluated the biological effects of Serenoa repens extract (Prostasan®) on prostate cells beyond its known antiandrogenic actions. Prostasan® inhibited epidermal growth factor (EGF) and lipopolysaccharide (LPS) induced proliferation of the prostatic epithelial, androgen independent cell line PC-3. At effective concentrations of 50 µg/mL, Prostasan® partly displaced EGF from EGF receptor (EGFR) but fully blocked EGF-induced cell proliferation of PC-3 cells. Similarly, Prostasan® inhibited LPS-induced proliferation of PC-3 cells without affecting LPS activation of the NFĸB pathway via toll-like receptor-4 (TLR-4). Additionally, Prostasan® reduced the constitutive secretion of monocyte chemotactic protein-1 (MCP-1), the LPS-induced secretion of IL-12 and inhibited MCP-1 and granulocyte-macrophage colony-stimulating factor (GM-CSF) production in the presence of LPS on PC-3 cells. Taken together, our results suggest that S. repens extracts, in addition to other reported effects on BPH development and prostatitis, inhibits EGF-dependent growth and proinflammatory responses of the prostate epithelial cells.

Topics: Cell Line; Cell Proliferation; Cytokines; Epidermal Growth Factor; Epithelial Cells; ErbB Receptors; Humans; Inflammation; Lipopolysaccharides; Male; Plant Extracts; Prostate; Serenoa

Ischemia-reperfusion (IR) injury is a common cause of acute renal failure. Bone marrow (BM)-derived mesenchymal stromal cells (MSC) delivered after renal IR are renoprotective, but knowledge of the protective mechanism is still in development. This investigation analyzed the protective molecular mechanisms of MSC, in particular relating to modulated oxidative stress.. In vivo and in vitro models of renal IR were analyzed with and without MSC. In vivo, adult male Sprague-Dawley rats were subjected to 40-min unilateral renal IR. Rat BM-derived MSC were administered at 24 h post-IR (IR + MSC). Other groups had IR but no MSC, or MSC but no ischemia (all groups n = 4). Apoptosis, inflammation, oxidative stress and reparative signal transduction molecules or growth factors were studied 4 days post-IR. In vitro, protection by MSC against oxidative stress (0.4 mm hydrogen peroxide) was investigated using rat renal tubular epithelial cells (NRK52E) with or without MSC in co-culture (tissue culture trans-well inserts), followed by similar analyses to the in vivo investigation.. In vivo, kidneys of IR + MSC animals had significantly increased cell proliferation/regeneration (cells positive for proliferating cell nuclear antigen, expression of epidermal growth factor), increased heme-oxygenase-1 (improved cell survival, anti-oxidant) and decreased 8-OHdG (decreased oxidative stress). In vitro, MSC delivered with oxidative stress significantly decreased apoptosis and Bax (pro-apoptotic protein), and increased mitosis and phospho-ERK1/2, thereby minimizing the damaging outcome and maximizing the regenerative effect after oxidative stress.. The benefits of MSC, in IR, were primarily pro-regenerative, sometimes anti-apoptotic, and novel anti-oxidant mechanisms were identified.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Bone Marrow Cells; Cell Line; Cell Proliferation; Epidermal Growth Factor; Gene Expression; Heme Oxygenase-1; Inflammation; Male; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Oxidative Stress; Proliferating Cell Nuclear Antigen; Rats; Rats, Sprague-Dawley; Regeneration; Reperfusion Injury; Signal Transduction

The epidermal growth factor receptor (EGFr) regulates many cellular functions, such as proliferation, apoptosis, and ion transport. Our aim was to investigate whether long term treatment with interferon-γ (IFN-γ) modulates EGF activation of downstream signaling pathways in intestinal epithelial cells and if this contributes to dysregulation of epithelial ion transport in inflammation. Polarized monolayers of T(84) and HT29/cl.19A colonocytes were preincubated with IFN-γ prior to stimulation with EGF. Basolateral potassium transport was studied in Ussing chambers. We also studied inflamed colonic mucosae from C57BL/6 mice treated with dextran sulfate sodium or mdr1a knock-out mice and controls. IFN-γ increased intestinal epithelial EGFr expression without increasing its phosphorylation. Conversely, IFN-γ caused a significant decrease in EGF-stimulated phosphorylation of specific EGFr tyrosine residues and activation of ERK but not Akt-1. In IFNγ-pretreated cells, the inhibitory effect of EGF on carbachol-stimulated K(+) channel activity was lost. In inflamed colonic tissues, EGFr expression was significantly increased, whereas ERK phosphorylation was reduced. Thus, although it up-regulates EGFr expression, IFN-γ causes defective EGFr activation in colonic epithelial cells via reduced phosphorylation of specific EGFr tyrosine residues. This probably accounts for altered downstream signaling consequences. These observations were corroborated in the setting of colitis. IFN-γ also abrogates the ability of EGF to inhibit carbachol-stimulated basolateral K(+) currents. Our data suggest that, in the setting of inflammation, the biological effect of EGF, including the inhibitory effect of EGF on Ca(2+)-dependent ion transport, is altered, perhaps contributing to diarrheal and other symptoms in vivo.

Topics: Animals; Carbachol; Cell Line; Diarrhea; Epidermal Growth Factor; Epithelial Cells; ErbB Receptors; Gene Expression Regulation; Humans; Inflammation; Interferon-gamma; Intestinal Mucosa; Ion Transport; Mice; Mice, Knockout; Miotics; Potassium; Proto-Oncogene Proteins c-akt

Immune inflammatory processes in prenatal and perinatal stages are suggested to play crucial roles in the vulnerability to schizophrenia. Based upon this immune inflammatory hypothesis for schizophrenia, we have established animal models for this illness by subcutaneously administering cytokines or proinflammatory agents to rodent neonates. These models exhibit various schizophrenia-like behavioral abnormalities after puberty, most of which are sensitive to various antipsychotics. The experimental procedures are all simple and easily utilized by researchers unfamiliar with these models. The behavioral changes are reproducible and remarkable but do not accompany learning deficits. The molecular and cellular targets of these agents have also been investigated and partially characterized, such as the cortical GABAergic system, midbrain dopaminergic system and hippocampal glutamate system. In this chapter, we introduce the details of the procedure and discuss the potential application of these animal models to drug development for schizophrenia.

Topics: Animals; Behavior, Animal; Cytokines; Disease Models, Animal; Epidermal Growth Factor; Female; GABAergic Neurons; Inflammation; Inflammation Mediators; Mesencephalon; Mice; Neuregulin-1; Poly I-C; Pregnancy; Prenatal Exposure Delayed Effects; Rats; Schizophrenia

Despite the excellent anti-inflammatory and immunosuppressive action of glucocorticoids (GCs), their use for the treatment of inflammatory bowel disease (IBD) still carries significant risks in terms of frequently occurring severe side effects, such as the impairment of intestinal tissue repair. The recently-introduced selective glucocorticoid receptor (GR) agonists (SEGRAs) offer anti-inflammatory action comparable to that of common GCs, but with a reduced side effect profile.. The in vitro effects of the non-steroidal SEGRAs Compound A (CpdA) and ZK216348, were investigated in intestinal epithelial cells and compared to those of Dexamethasone (Dex). GR translocation was shown by immunfluorescence and Western blot analysis. Trans-repressive effects were studied by means of NF-κB/p65 activity and IL-8 levels, trans-activation potency by reporter gene assay. Flow cytometry was used to assess apoptosis of cells exposed to SEGRAs. The effects on IEC-6 and HaCaT cell restitution were determined using an in vitro wound healing model, cell proliferation by BrdU assay. In addition, influences on the TGF-β- or EGF/ERK1/2/MAPK-pathway were evaluated by reporter gene assay, Western blot and qPCR analysis.. Dex, CpdA and ZK216348 were found to be functional GR agonists. In terms of trans-repression, CpdA and ZK216348 effectively inhibited NF-κB activity and IL-8 secretion, but showed less trans-activation potency. Furthermore, unlike SEGRAs, Dex caused a dose-dependent inhibition of cell restitution with no effect on cell proliferation. These differences in epithelial restitution were TGF-β-independent but Dex inhibited the EGF/ERK1/2/MAPK-pathway important for intestinal epithelial wound healing by induction of MKP-1 and Annexin-1 which was not affected by CpdA or ZK216348.. Collectively, our results indicate that, while their anti-inflammatory activity is comparable to Dex, SEGRAs show fewer side effects with respect to wound healing. The fact that SEGRAs did not have a similar effect on cell restitution might be due to a different modulation of EGF/ERK1/2 MAPK signalling.

Topics: Active Transport, Cell Nucleus; Anti-Inflammatory Agents, Non-Steroidal; Apoptosis; Benzofurans; Benzoxazines; Cell Line; Cell Nucleus; Cell Proliferation; Epidermal Growth Factor; Gene Silencing; Humans; Inflammation; Intestinal Mucosa; MAP Kinase Signaling System; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Receptors, Glucocorticoid; Transcription, Genetic; Transcriptional Activation; Transforming Growth Factor beta; Wound Healing

In adults, circulating inflammatory mediators and activated CD14(++) monocytes link obesity to its metabolic and cardiovascular complications. However, it is largely unknown whether these inflammatory changes already occur in childhood obesity. To survey inflammatory changes during the early stages of obesity, we performed a comprehensive analysis of circulating inflammatory mediators, monocyte populations and their function in childhood obesity.. In lean and obese children aged 6 to 16 years (n = 96), 35 circulating inflammatory mediators including adipokines were measured. Hierarchical cluster analysis of the inflammatory mediator profiles was performed to investigate associations between inflammatory mediator clusters and clinical variables. Whole-blood monocyte phenotyping and functional testing with the toll-like receptor 4 ligand, lipopolysaccharide, were also executed.. First, next to leptin, the circulating mediators chemerin, tissue inhibitor of metalloproteinase 1, EGF and TNF receptor 2 were identified as novel inflammatory mediators that are increased in childhood obesity. Second, cluster analysis of the circulating mediators distinguished two obesity clusters, two leanness clusters and one mixed cluster. All clusters showed distinct inflammatory mediator profiles, together with differences in insulin sensitivity and other clinical variables. Third, childhood obesity was associated with increased CD14(++) monocyte numbers and an activated phenotype of the CD14(++) monocyte subsets.. Inflammatory mediator clusters were associated with insulin resistance in obese and lean children. The activation of CD14(++) monocyte subsets, which is associated with increased development of atherosclerosis in obese adults, was also readily detected in obese children. Our results indicate that inflammatory mechanisms linking obesity to its metabolic and cardiovascular complications are already activated in childhood obesity.

Topics: Adolescent; Case-Control Studies; Cell Count; Chemokines; Child; Cluster Analysis; Comorbidity; Epidermal Growth Factor; Female; Humans; Inflammation; Inflammation Mediators; Intercellular Signaling Peptides and Proteins; Leptin; Lipopolysaccharide Receptors; Male; Monocytes; Obesity; Receptors, Tumor Necrosis Factor, Type II; Regression Analysis; Tissue Inhibitor of Metalloproteinase-1; Vascular Endothelial Growth Factor A

Our goal was to assess the toxicity of two strengths (200 and 400 μg) of HER1 cancer vaccine (Center of Molecular Immunology, Cuba), presented in two different formulations, in Sprague Dawley rats after repeated intramuscular administration (14 days). Four groups (5 animals/sex) were established: Control, Placebo (adjuvant), and two Treated groups receiving a dose representing ten times of human total dose (10×), 28.6 and 57.1 μg/kg. Clinical observations, body weight and rectal temperature were measured during the study. Clinical pathology analysis was performed, besides gross necropsy and histological examination of tissues on animals at the end of the assay. The assay ended with a 100% survival. Injection site damage, with the presence of cysts and granulomas, was observed in adjuvant and vaccine treated groups, with most severe cases predominating at higher strength. Administration of Placebo and Her1 vaccine induced increase in polymorphonuclear cells, with relative lymphopenia conditioned by primary neutrophilia. In summary, results suggest that Her1 immunization was capable of inducing an inflammatory effect at the injection site, leading to systemic alterations, more significant at higher strength (400 μg, 57.1 μg/kg), probably affected by the immunizations' schedule used. The vaccine was shown to be well tolerated without any obvious signs of systemic toxicity, with findings largely attributable to the adjuvant used.

Topics: Animals; Cancer Vaccines; Dose-Response Relationship, Drug; Drug Administration Schedule; Epidermal Growth Factor; Female; Inflammation; Injections, Intramuscular; Male; Neutrophils; Rats; Rats, Sprague-Dawley

CD40, a co-stimulatory molecule, plays a critical role in coordinating enteric inflammatory immune responses. In necrotizing enterocolitis (NEC), upregulation of IL-10, a CD40-modulated cytokine, has been described, but the role of the IL-10 receptor (IL-10Rβ), CD40, and its ligand CD40L in disease pathogenesis is unknown. The study herein investigates ileal expression of CD40, CD40L, and IL-10R in a rat model of NEC.. NEC was induced in newborn rats using established methods of formula feeding, asphyxia, and cold stress. Expression of CD40, CD40L, IL-10Rβ, and other proinflammatory molecules, including Toll-like receptor-4 (TLR-4) and IL-18, was assessed by immunoblotting. Tissue infiltration by macrophages, monocytes, and T cells was examined by confocal immunohistochemistry.. Ileum from rat pups with NEC showed increased expression of TLR-4, IL-18, and IL-10Rβ. Sections from both NEC and control pups demonstrated preservation of ileal cells expressing CD40/CD40L. The distal ileum of controls expressed both CD40 and CD40L; conversely, neither molecule was detected in ileal tissue from NEC pups. Additional studies showed that treatment with epidermal growth factor (EGF), previously shown to ameliorate the severity of NEC in an animal model, did not restore CD40 expression.. Ileal cytokine dysregulation, manifested by decreased CD40/CD40L and increased IL-10Rβ expression, may be involved in the pathogenesis of NEC. Dampened CD40 signaling may be related to enhanced IL-10 expression and a suppressed T-cell response to injury. We speculate that augmenting CD40-CD40L interactions may achieve a protective effect in this NEC model.

Topics: Animals; Blotting, Western; CD40 Antigens; CD40 Ligand; Enterocolitis, Necrotizing; Epidermal Growth Factor; Ileum; Inflammation; Interleukin-10 Receptor beta Subunit; Interleukin-18; Macrophages; Models, Animal; Monocytes; Rats; Rats, Sprague-Dawley; T-Lymphocytes; Toll-Like Receptor 4

To investigate effects of hepatotropic growth factors on radical production in rat hepatocytes during sepsis.. Rat hepatocytes, isolated by collagenase perfusion, were incubated with a lipopolysaccharide (LPS)-containing cytokine mixture of interleukin-1β, tumor necrosis factor-α and interferon-γ to simulate sepsis and either co-incubated or pre-incubated with hepatotropic growth factors, e.g. hepatocyte growth factor, epidermal growth factor and/or transforming growth factor-α. Cells were analyzed for glutathione levels. Culture supernatants were assayed for production of reactive oxygen intermediates (ROIs) as well as NO(2) (-), NO(3) (-) and S-nitrosothiols. To determine cellular damage, release of aspartate aminotransferase (AST) into the culture medium was analyzed. Activation of nuclear factor (NF)-κB was measured by electrophoretic mobility shift assay.. Rat hepatocytes treated with the LPS-containing cytokine mixture showed a significant increase in ROI and nitrogen oxide intermediate formation. AST leakage was not significantly increased in cells treated with the LPS-containing cytokine mixture, independent of growth-factor co-stimulation. However, pretreatment with growth factors significantly reduced AST leakage and ROI formation while increasing cellular glutathione. Application of growth factors did not result in increased NF-κB activation. Pretreatment with growth factors further increased formation of NO(2) (-), NO(3) (-) and S-nitrosothiols in hepatocytes stimulated with LPS-containing cytokine mixture. Thus, we propose that, together with an increase in glutathione increased NO(2) (-), NO(3) (-) formation might shift their metabolism towards non-toxic products.. Our data suggest that hepatotropic growth factors positively influence sepsis-induced hepatocellular injury by reducing cytotoxic ROI formation via induction of the cellular protective antioxidative systems.

Topics: Animals; Antioxidants; Epidermal Growth Factor; Glutathione; Hepatocyte Growth Factor; Hepatocytes; Inflammation; NF-kappa B; Nitric Oxide; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Transforming Growth Factor alpha; Up-Regulation

iNKT cells are a unique T cell subset, which is CD1d-restricted and specific for glycolipid antigens. In advanced atherosclerotic plaques, focal collections of inflammatory cells correlate with areas of intraplaque neovascularization. We reported recently that iNKT cells might facilitate intraplaque neovascularization by enhancing EC migration and sprouting in an IL-8-dependent manner. This study investigated the participating effector mechanisms. In ECs, CM, derived from antigen-stimulated human iNKT cells (CM+), induced up-regulation of IL-8R CXCR2 and the phosphorylation of EGFR and of multiple intracellular signaling effectors, including FAK, Src, Erk, Jnk, p38-MAPK, and STAT1 and -3. We found that a cascade of events, which were IL-8-dependent and involved EGFR activation, was responsible for signaling through FAK and Src kinases and necessary for acquisition of angiogenic morphology, migration in a two-dimensional wound assay, and sprout outgrowth in a three-dimensional model of angiogenesis in vitro. The data support that IL-8-dependent activation of angiogenic behavior in ECs, in response to activated iNKT, involves CXCR2, transactivation of EGFR, and subsequent FAK/Src signaling. We found too that activated iNKT increased VEGFR2 expression in ECs. Functional studies confirmed that EGF is the motogenic-enhancing factor in CM+ and is necessary, together with an exogenous source of VEGF, for iNKT-promoted sprout formation. EGFR inhibition may represent a novel therapeutic modality aimed at plaque stabilization through control of neovascularization within developing atherosclerotic plaques.

Topics: Antigens; Atherosclerosis; Cell Communication; Cell Line; Cell Movement; Endothelial Cells; Epidermal Growth Factor; ErbB Receptors; Humans; Inflammation; Interleukin-8; Lipids; Lymphocyte Activation; Natural Killer T-Cells; Neovascularization, Pathologic; Phosphorylation; Receptors, Interleukin-8B; Signal Transduction; Up-Regulation

Topics: Animals; Cicatrix; Epidermal Growth Factor; Fibrosis; Humans; Inflammation; Interferon-gamma; Interleukin-10; Ligands; Mice; Models, Biological; Receptors, CXCR3; Signal Transduction; Stress, Mechanical; Wound Healing

A visible cutaneous scar develops from the excess formation of immature collagen in response to an inflammatory reaction. This study examined the role of epidermal growth factor (EGF) in the formation of cutaneous scars. Twenty Crl:CD-1 (ICR) mice were used and 2 full-thickness skin wounds were made on the dorsum of each mouse. One of the wounds was treated with recombinant human EGF by local application and the other was treated with saline for control until complete healing was achieved. The EGF-treated group's wounds healed faster than the control group's. The width of the scar was smaller by 30% and the area was smaller by 26% in the EGF-treated group. Inflammatory cell numbers were significantly lower in the EGF-treated group. The expression of transforming growth factor (TGF)-beta(1) in the EGF-treated group was increased. It was observed that the amount of collagen in the EGF-treated group was larger than the control group. In the EGF-treated group, the visible external scars were less noticeable than that in the control group. These results suggest that EGF can reduce cutaneous scars by suppressing inflammatory reactions, decreasing expression of TGF-beta(1), and mediating the formation of collagen.

Topics: Animals; Cicatrix; Collagen; Epidermal Growth Factor; Humans; Inflammation; Mice; Recombinant Proteins; Skin; Wound Healing

Inhibition of inflammatory responses, acceleration of basal cell growth and balanced synthesis of the extracellular matrix (ECM) are important in the healing of open cutaneous wounds. To evaluate the wound-healing effects of Astragali Radix (AR) (the root of Astragalus membranaceus [Fisch.]), experimental open wounds were made on the dorsal side of rats under anesthesia. Boiling water extracts of AR, soaked into a hydrophilic foam dressing, were topically applied to the wounds once a day for 11 consecutive days. The healing process was assessed by scoring macroscopic appearance and measuring the area of the open wounds. Molecular aspects of the healing skin area were also investigated via histological observation indicating cell density and linear alignment of the granulation tissue. The AR extracts significantly accelerated cutaneous wound healing by suppressing inflammation and stimulating basal cell growth in the wound area compared to epidermal growth factor as a positive control. Promotion of basal cell proliferation and angiogenesis by the AR extracts was remarkable in the early stages of wound healing, resulting in a significant reduction in the duration of the wound-healing process. We conclude that extracts of AR could be useful in enhancing cutaneous wound healing.

Topics: Administration, Cutaneous; Animals; Astragalus propinquus; Cell Proliferation; Disease Models, Animal; Epidermal Growth Factor; Inflammation; Male; Neovascularization, Physiologic; Plant Extracts; Plant Roots; Rats; Rats, Sprague-Dawley; Skin; Wound Healing

Psychosocial stress is becoming a major contributor to increased mental ill-health and sick leave in many countries. Valid markers of chronic stress would be valuable for diagnostic and prognostic purposes. A recent study suggested monocyte chemotactic protein-1 (MCP-1), epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) as markers of chronic stress. We aimed to confirm these potential biomarkers of prolonged psychosocial stress in female patients.. Circulating levels of MCP-1, EGF and VEGF, along with several other cytokines, were measured in plasma from 42 female patients suffering from exhaustion due to prolonged psychosocial stress and 42 control subjects, using a protein biochip immunoassay. There were no significant differences between patients and controls in any of the cytokines or growth factors analyzed. Furthermore, when using a different protein bioassay and reanalyzing MCP-1 and VEGF in the same samples, markedly different levels were obtained. To further explore if inflammation is present in patients with exhaustion, the classical inflammatory marker C-reactive protein (CRP) was measured in another group of patients (n=89) and controls (n=88) showing a small but significant increase of CRP levels in the patients.. MCP-1, EGF and VEGF may not be suitable markers of prolonged psychosocial stress as previously suggested. Furthermore, significant differences were obtained when using two different protein assays measuring the same samples, indicating that comparing studies where different analytic techniques have been used might be difficult. Increased levels of CRP indicate that low-grade inflammation might be present in patients with exhaustion due to prolonged stress exposure but this inflammation does not seem to be reflected by increase in circulating MCP-1 or other cytokines measured.

Topics: Adult; Biological Assay; Biomarkers; C-Reactive Protein; Case-Control Studies; Chemokine CCL2; Epidermal Growth Factor; Female; Humans; Inflammation; Middle Aged; Protein Array Analysis; Stress, Psychological; Surveys and Questionnaires; Vascular Endothelial Growth Factor A

Exercise is thought to stimulate the release of hematopoietic and endothelial progenitor cells (EPC) from the bone marrow. Little is known about the influence of strenuous exercise on the content of circulating progenitor cells. The aim of this study was to investigate the influence of a marathon race on the amount of circulating progenitor cells immediately after the race in advanced-aged runners.. Sixty-eight healthy marathon runners (age: 57+/-6 years) were included in this study. Blood cell counts were evaluated by standard methods, and circulating progenitor cells before and immediately after the race were quantified by fluorescence-activated cell sorter (FACS). Vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) was quantified by enzyme-linked immunosorbent assay.. A marathon race led to a significant increase in white blood cell count (5283+/-155 vs. 13706+/-373 cells/mul; P<0.001). Fluorescence-activated cell sorter analysis revealed a significant decrease of CD34 cells (1829+/-115 vs. 1175+/-75 cells/ml blood; P<0.0001), CD117 cells (2478+/-245 vs. 2193+/-85 cells/ml blood; P<0.05), and CD133 cells (3505+/-286 vs. 2239+/-163 cells/ml blood; P<0.001). No significant change was observed for EPCs defined as CD34/VEGF-R2 cells (117+/-8 vs. 128+/-9 cells/ml blood; P=0.33). With respect to VEGF a significant downregulation was evident directly after the race (48.9+/-8.0 vs. 34.0+/-7.5 pg/ml; P<0.05), whereas no change was obvious in EGF levels.. The results of our study suggest that finishing a marathon race will lead to an inflammatory response and downregulation of circulating hematopoietic stem cells. With respect to EPCs no change is observed, which may be because of a greater differentiation of the remaining CD34 cells towards EPCs.

Topics: AC133 Antigen; Age Factors; Antigens, CD; Antigens, CD34; Down-Regulation; Endothelial Cells; Epidermal Growth Factor; Glycoproteins; Hematopoietic Stem Cells; Humans; Inflammation; Leukocyte Count; Middle Aged; Peptides; Physical Endurance; Proto-Oncogene Proteins c-kit; Running; Stem Cells; Vascular Endothelial Growth Factor A

Acidic mammalian chitinase (AMCase) is expressed in an exaggerated fashion in epithelial cells at sites of pulmonary T helper cell type 2 inflammation and plays important roles in the pathogenesis of anti-parasite and asthma-like responses. However, the mechanisms that control epithelial cell AMCase secretion and its effector responses have not been adequately defined. To address these issues, we used in vivo and in vitro experimental systems to define the pathways of epithelial AMCase secretion and its epithelial regulatory effects. Here we demonstrate that, in murine T helper cell type 2 modeling systems, AMCase colocalizes with the epidermal growth factor receptor (EGFR) and ADAM17 (a membrane disintegrin and metallopeptidase 17) in lung epithelial cells. In vitro cotransfection experiments in A549 cells demonstrated that AMCase and EGFR physically interact with each other. Cotransfection of AMCase and EGFR also increased, whereas EGFR inhibition decreased AMCase secretion. Interestingly, AMCase secretion was not significantly altered by treatment with EGF but was significantly decreased when the upstream EGFR transactivator ADAM17 was inhibited. AMCase secretion was also decreased when the EGFR-downstream Ras was blocked. Transfected and recombinant AMCase induced epithelial cell production of CCL2, CCL17, and CXCL8. These studies demonstrate that lung epithelial cells secrete AMCase via an EGFR-dependent pathway that is activated by ADAM17 and mediates its effects via Ras. They also demonstrate that the AMCase that is secreted feeds back in an autocrine and/or paracrine fashion to stimulate pulmonary epithelial cell chemokine production.

Topics: ADAM Proteins; ADAM17 Protein; Animals; Cell Line; Chemokines; Chitinases; Epidermal Growth Factor; Epithelial Cells; ErbB Receptors; Humans; Inflammation; Lung; Mice; Mice, Transgenic; ras Proteins; Respiratory Mucosa; Th2 Cells; Transfection

The bronchial epithelium is an important physical barrier that regulates physiological processes including leukocyte trafficking. The aim of the present study was to elucidate the mechanisms whereby the bronchial epithelium, stimulated by epidermal growth factor (EGF) as part of a response to acute or chronic injury, could activate and chemoattract human neutrophils. Subconfluent human bronchial epithelial (16HBE) cells were stimulated with EGF to mimic the in vivo events after injury. The effect of the resulting EGF-conditioned media (CM) was compared with that of basal-CM with respect to neutrophil activation and chemotaxis. Such findings were then confirmed using primary bronchial epithelial cells (PBECs) from healthy volunteers. EGF-CM from 16HBE cells caused increased expression of CD11b/CD66b and CD62L loss on neutrophils when compared with basal-CM. EGF-CM contained significant neutrophil chemotactic activity involving granulocyte-macrophage colony-stimulating factor and interleukin-8 that was potentiated by leukotriene B(4). This was dependent on neutrophil phosphatidylinositol-3-kinase activation and Akt phosphorylation, with partial regulation by phospholipase D, but not mammalian target of rapamycin. Consistent with these observations, EGF-CM derived from PBECs displayed increased chemotactic activity. The present results suggest that the enhanced chemotactic activity of the epidermal growth factor-conditioned epithelium can enhance neutrophil-mediated immunity during acute injury, while during continued injury and repair, as in chronic asthma, this could contribute to persistent neutrophilic inflammation.

Topics: Bronchi; Cell Line; Chemotaxis, Leukocyte; Culture Media, Conditioned; Epidermal Growth Factor; Epithelial Cells; Humans; Inflammation; Neutrophils; Respiratory Distress Syndrome; Signal Transduction

Exposure of adult humans to manganese (Mn) has long been known to cause neurotoxicity. Recent evidence also suggests that exposure of children to Mn is associated with developmental neurotoxicity. Astrocytes are critical for the proper functioning of the nervous system, and they play active roles in neurogenesis, synaptogenesis and synaptic neurotransmission. In this report, to help elucidate the molecular events underlying Mn neurotoxicity, we systematically identified the molecular targets of Mn in primary human astrocytes at a genome-wide level, by using microarray gene expression profiling and computational data analysis algorithms. We found that Mn altered the expression of diverse genes ranging from those encoding cytokines and transporters to signal transducers and transcriptional regulators. Particularly, 28 genes encoding proinflammatory chemokines, cytokines and related functions were up-regulated, whereas 15 genes encoding functions involved in DNA replication and repair and cell cycle checkpoint control were down-regulated. Consistent with the increased expression of proinflammatory factors, analysis of common regulators revealed that 16 targets known to be positively affected by the interferon-gamma signaling pathway were up-regulated by Mn(2+). In addition, 68 genes were found to be similarly up- or down-regulated by both Mn(2+) and hypoxia. These results from genomic analysis are further supported by data from real-time RT-PCR, Western blotting, flow cytometric and toxicological analyses. Together, these analyses show that Mn(2+) selectively affects cell cycle progression, the expression of hypoxia-responsive genes, and the expression of proinflammatory factors in primary human astrocytes. These results provide important insights into the molecular mechanisms underlying Mn neurotoxicity.

Topics: Astrocytes; Blotting, Western; Carrier Proteins; Cell Cycle; Cell Hypoxia; Cells, Cultured; Chlorides; DNA Repair; DNA Replication; Epidermal Growth Factor; Flow Cytometry; Gene Expression; Gene Expression Profiling; Humans; Immunity, Cellular; Inflammation; Interferon-gamma; L-Lactate Dehydrogenase; Manganese Compounds; Oligonucleotide Array Sequence Analysis; Reverse Transcriptase Polymerase Chain Reaction; RNA; Signal Transduction; Transcription, Genetic; Up-Regulation

During vascular disease and following injury, vascular smooth muscle cells (VSMC) proliferate and produce inflammation-promoting cytokines and chemokines. Similar phenotypic changes can be elicited in vitro by activation of Toll-like receptors (TLR) within VSMC. TLR-activated VSMC also produce IL-1 alpha, but it is unknown whether endogenous IL-1 alpha stimulates VSMC in an autocrine manner. Here we tested the hypothesis that endogenous IL-1 alpha contributes to TLR-induced proliferation and chemokine release in human VSMC by using RNA interference to knock down IL-1 alpha expression. Knockdown of IL-1 alpha abolished TLR-induced proliferation and suppressed TLR4-induced release of monocyte chemoattractant protein-1 (MCP-1) by VSMC, indicating that endogenous IL-1 alpha plays a crucial role in both responses. Serum, PDGF, FGF-2, and EGF each increased cellular IL-1 alpha concentrations, and IL-1 alpha knockdown inhibited serum- and PDGF-induced DNA synthesis, further indicating that endogenous IL-1 alpha also contributed to VSMC responses to growth factors. IL-1 receptor antagonist, a competitive inhibitor of IL-1 receptor I (IL-1RI), also attenuated TLR-induced proliferation and both basal and TLR-induced MCP-1 expression, indicating at least a partial role of the IL-1RI in mediating these responses. The results support the hypothesis that autocrine actions of endogenous IL-1 alpha, mediated at least in part via IL-1RI signaling, contribute to a proproliferative and proinflammatory phenotypic shift in TLR-activated human VSMC, which might play a pathogenic role in vascular disorders.

Topics: Autocrine Communication; Cell Proliferation; Cells, Cultured; Chemokine CCL2; Coronary Vessels; Epidermal Growth Factor; Fibroblast Growth Factor 2; Humans; Inflammation; Interleukin 1 Receptor Antagonist Protein; Interleukin-1alpha; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Phenotype; Platelet-Derived Growth Factor; Receptors, Interleukin-1 Type I; RNA Interference; RNA, Small Interfering; Serum; Toll-Like Receptor 3; Toll-Like Receptor 4; Toll-Like Receptors; Transfection

Although high-sensitivity C-reactive protein (hs-CRP) has emerged as a cardiovascular marker, questions arise regarding the relative information provided by other inflammatory molecules. Therefore, as a first step, we examined interrelationships between serum hs-CRP concentrations and inflammatory, adhesion and growth factors in healthy adults.. Circulating concentrations of hs-CRP, haptoglobin, orosomucoid, interleukin-6 (IL-6), IL-8, IL-18, tumor necrosis factor-alpha (TNF-alpha), TNF-receptor II (TNF-RII), E-, P-, and L-selectins, intercellular adhesion molecule-1 (ICAM-1), monocyte chemoattractant protein-1, endothelial growth factor (EGF), vascular EGF (VEGF), insulin-like growth factor-1 (IGF-1) and IGF-binding protein (IGFBP-3) were measured in 154 men and 161 women of the Stanislas cohort. Leukocyte and platelet counts were also determined.. Correlations were significant between hs-CRP concentrations and leukocyte and platelet counts, as well as haptoglobin, orosomucoid, IL-6, and ICAM-1 concentrations (p< or =0.001). Correlation coefficients for ICAM-1 were higher in men than in women (p< or =0.05). When stratifying subjects according to hs-CRP levels, the group with high hs-CRP levels had significantly higher haptoglobin and orosomucoid concentrations than the others, in addition to higher leukocyte counts and IL-6 concentrations in women, and platelet counts and ICAM-1 concentrations in men.. Further studies are warranted to explain the association pattern for hs-CRP. Partition of these factors according to their association with hs-CRP concentration opens a new perspective for choice of the best factors in terms of cardiovascular risk in relation to hs-CRP, while non-associated markers could be used to give additional information.

Topics: Adult; Biomarkers; Blood Cell Count; C-Reactive Protein; Child, Preschool; Cohort Studies; Epidermal Growth Factor; Female; Haptoglobins; Humans; Inflammation; Insulin-Like Growth Factor Binding Protein 3; Insulin-Like Growth Factor I; Intercellular Adhesion Molecule-1; Interleukins; Male; Middle Aged; Orosomucoid; Selectins; Tumor Necrosis Factors; Vascular Endothelial Growth Factor A

A complex network of regulatory neuropeptides controls airway inflammation reaction, in which airway epithelial cells adhering to and activating leukocytes is a critical step. To study the effect of intrapulmonary regulatory peptides on adhesion of polymorphonuclear leukocytes (PMNs) to bronchial epithelial cells (BECs) and its mechanism, several regulatory peptides including vasoactive intestinal peptide (VIP), epidermal growth factor (EGF), endothelin-1 (ET-1) and calcitonin gene-related peptide (CGRP), were investigated. The results demonstrated that VIP and EGF showed inhibitory effects both on the secretion of IL-1, IL-8 and the adhesion of PMNs to BECs, whereas ET-1 and CGRP had the opposite effect. Anti-intercellular adhesion molecule-1 (ICAM-1) antibody could block the adhesion of PMNs to ozone-stressed BECs. Using immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR), it was shown that VIP and EGF down-regulated the expression of ICAM-1 in BECs, while ET-1 and CGRP up-regulated ICAM-1 expression. NF-kappaB inhibitor MG132 blocked ICAM-1 expression induced by ET-1 and CGRP. Furthermore, in electric mobility shift assay (EMSA), VIP and EGF restrained the binding activity of NF-kappaB to the NF-kappaB binding site within the ICAM-1 promoter in ozone-stressed BECs, while CGRP and ET-1 promoted this binding activity. IkappaB degradation was consistent with NF-kappaB activation. These observations indicate that VIP and EGF inhibit inflammation, while ET-1 and CGRP enhance the inflammation reaction.

Topics: Animals; Base Sequence; Bronchi; Calcitonin Gene-Related Peptide; Cell Adhesion; Cells, Cultured; Endothelin-1; Epidermal Growth Factor; Epithelial Cells; I-kappa B Kinase; Inflammation; Intercellular Adhesion Molecule-1; Interleukin-1; Interleukin-8; Interleukins; Neutrophils; NF-kappa B; Peptides; Rabbits; Time Factors; Vasoactive Intestinal Peptide

The intensity of neutrophil inflammatory response could be rapidly amplified by priming with pro-inflammatory mediators such as TNF-alpha, GM-CSF or LPS at low concentrations prior to stimuli. We proposed that epidermal growth factor (EGF) increases TNF-alpha-induced priming of human neutrophils. This study showed that EGF enhanced TNF-alpha-induced activation of neutrophils functions. The addition of EGF to neutrophils cultured with TNF-alpha resulted in increased respiratory burst and phagocytic activity of polymorphonuclear leukocytes (PMN) and up-regulation of adhesion molecule CD11b. Moreover, EGF enhanced IL-8 production by TNF-alpha-primed PMN. EGF alone was able to prime CD11b expression and IL-8 production by PMN. EGF receptor selective tyrosine kinase inhibitor, tyrphostin AG-1517, blocked the effect of priming with EGF, whereas the status of non-primed and TNF-alpha-primed neutrophils remained unaffected. EGFR expression on neutrophils was confirmed by flow cytometry and CELISA methods. These data provide the original evidence that EGF significantly enhances TNF-alpha-induced priming of human neutrophils acting through EGFR tyrosine kinase pathway. The observed effect may be a result of co-operative action of EGF, TNF-alpha and reactive oxygen intermediates (ROI).

Topics: CD11b Antigen; Cells, Cultured; Epidermal Growth Factor; ErbB Receptors; Humans; Inflammation; Interleukin-8; Neutrophils; Phagocytosis; Protein Kinase Inhibitors; Quinazolines; Reactive Oxygen Species; Respiratory Burst; Tumor Necrosis Factor-alpha; Tyrphostins

Cardiac hypertrophy is a complex and nonhomogenous response to various stimuli. In this study, we used high-density oligonucleotide microarray to examine gene expression profiles during physiological hypertrophy, pathological hypertrophy, and heart failure in Dahl salt-sensitive rats. There were changes in 404/3,160 and 874/3,160 genes between physiological and pathological hypertrophy and the transition from hypertrophy to heart failure, respectively. There were increases in stress response genes (e.g., heat shock proteins) and inflammation-related genes (e.g., pancreatitis-associated protein and arachidonate 12-lipoxygenase) in pathological processes but not in physiological hypertrophy. Furthermore, atrial natriuretic factor and brain natriuretic protein showed distinctive changes that are very specific to different conditions. In addition, we used a resampling-based gene score-calculating method to define significantly altered gene clusters, based on Gene Ontology classification. It revealed significant alterations in genes involved in the apoptosis pathway during pathological hypertrophy, suggesting that the apoptosis pathway may play a role during the transition to heart failure. In addition, there were significant changes in glucose/insulin signaling, protein biosynthesis, and epidermal growth factor signaling during physiological hypertrophy but not during pathological hypertrophy.

Topics: Animals; Apoptosis; Atrial Natriuretic Factor; Blotting, Northern; Cardiomegaly; Echocardiography; Epidermal Growth Factor; Gene Expression Profiling; Gene Expression Regulation; Heart Failure; Hypertrophy; Inflammation; Insulin; Natriuretic Peptide, Brain; Oligonucleotide Array Sequence Analysis; Pancreatitis-Associated Proteins; Physical Conditioning, Animal; Rats; Rats, Inbred Dahl; RNA; Signal Transduction

Hepatic ischemia-reperfusion injury is an inevitable consequence during liver surgery. The outcome is particularly poor in cirrhotic livers, which are more prone to hepatic ischemia-reperfusion injury. We aim to study whether FTY720 could attenuate hepatic ischemia-reperfusion injury both in normal and in cirrhotic livers. We applied a 70% liver-ischemia (60 min) model in rats with normal or cirrhotic livers. FTY720 was given 20 min before ischemia and 10 min before reperfusion (1 mg/kg, i.v.). Liver tissues and blood were sampled at 20 min, 60 min, 90 min, 6 h and 24 h after reperfusion for detection of MAPK-Egr-1, Akt pathways and caspase cascade. Hepatic ultrastructure and apoptosis were also compared. FTY720 significantly improved liver function in the rats with normal and cirrhotic livers. Akt pathway was activated at 6 and 24 h after reperfusion. FTY720 significantly down-regulated Egr-1, ET-1, iNOS and MIP-2 accompanied with up-regulation of A20, IL-10, HO-1 and Hsp70. MAPK (Raf-MEK-Erk) pathway was down-regulated. Hepatic ultrastructure was well maintained and fewer apoptotic liver cells were found in the FTY720 groups. In conclusion, FTY720 attenuates ischemia-reperfusion injury in both normal and cirrhotic livers by activation of cell survival Akt signaling and down-regulation of Egr-1 via Raf-MEK-Erk pathway.

Topics: Animals; Apoptosis; Blotting, Western; Chemokine CXCL2; Chemokines, CXC; DNA Primers; Down-Regulation; Endothelin-1; Epidermal Growth Factor; Extracellular Signal-Regulated MAP Kinases; Fibrosis; Fingolimod Hydrochloride; Gene Expression Regulation; Heme Oxygenase (Decyclizing); Heme Oxygenase-1; Hepatocytes; HSP70 Heat-Shock Proteins; Immunosuppressive Agents; In Situ Nick-End Labeling; Inflammation; Intercellular Signaling Peptides and Proteins; Interleukin-10; Liver; Male; MAP Kinase Signaling System; Microscopy, Electron; Nitric Oxide Synthase; Nitric Oxide Synthase Type II; p38 Mitogen-Activated Protein Kinases; Propylene Glycols; Proteins; raf Kinases; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Sphingosine; Time Factors; Up-Regulation

Because matrix metalloproteinases (MMPs) play roles in inflammatory tissue injury, we asked whether MMP secretion by gastric epithelial cells may contribute to gastric injury in response to signals involved in Helicobacter pylori-induced inflammation and/or cyclooxygenase inhibition. Tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and epidermal growth factor (EGF) stimulated gastric cell MMP-1 secretion, indicating that MMP-1 secretion occurs in inflammatory as well as non-inflammatory situations. MMP-1 secretion required activation of the MAPK Erk and subsequent protein synthesis but was down-regulated by the alternate MAPK, p38. In contrast, secretion of MMP-13 was stimulated by TNF-alpha/IL-1beta but not EGF and was Erk-independent and mediated by p38. MMP-13 secretion was more rapid (peak, 6 h) than MMP-1 (peak > or =30 h) and only partly depended on protein synthesis, suggesting initial release of a pre-existing MMP-13 pool. Therefore, MMP-1 and MMP-13 secretion are differentially regulated by MAPKs. MMP-1 secretion was regulated by E prostaglandins (PGEs) in an Erk-dependent manner. PGEs enhanced Erk activation and MMP-1 secretion in response to EGF but inhibited Erk and MMP-1 when TNF-alpha and IL-1beta were the stimuli, indicating that the effects of PGEs on gastric cell responses are context-dependent. These data show that secretion of MMPs is differentially regulated by MAPKs and suggest mechanisms through which H. pylori infection and/or cyclooxygenase inhibition may induce epithelial cell signaling to contribute to gastric ulcerogenesis.

Topics: Cell Line, Tumor; Collagenases; Cycloheximide; Cyclooxygenase Inhibitors; Down-Regulation; Enzyme Activation; Enzyme Inhibitors; Epidermal Growth Factor; Epithelial Cells; Extracellular Signal-Regulated MAP Kinases; Gastric Mucosa; Gene Expression Regulation, Enzymologic; Helicobacter pylori; Humans; Inflammation; Interleukin-1; Kinetics; MAP Kinase Signaling System; Matrix Metalloproteinase 1; Matrix Metalloproteinase 13; Matrix Metalloproteinases; Models, Biological; p38 Mitogen-Activated Protein Kinases; Prostaglandins E; Protein Synthesis Inhibitors; Signal Transduction; Stomach; Time Factors; Tumor Necrosis Factor-alpha

The CCAAT/enhancer binding protein delta (C/EBPdelta, CRP3, CELF, NF-IL6beta) regulates gene expression and plays functional roles in many tissues, such as in acute phase response to inflammatory stimuli, adipocyte differentiation, and mammary epithelial cell growth control. In this study, we examined the expression of human C/EBPdelta (NF-IL6beta) gene by epidermal growth factor (EGF) stimulation in human epidermoid carcinoma A431 cells. NF-IL6beta was an immediate-early gene activated by the EGF-induced signaling pathways in cells. By using 5'-serial deletion reporter analysis, we showed that the region comprising the -347 to +9 base pairs was required for EGF response of the NF-IL6beta promoter. This region contains putative consensus binding sequences of Sp1 and cAMP response element-binding protein (CREB). The NF-IL6beta promoter activity induced by EGF was abolished by mutating the sequence of cAMP response element or Sp1 sites in the -347/+9 base pairs region. Both in vitro and in vivo DNA binding assay revealed that the CREB binding activity was low in EGF-starved cells, whereas it was induced within 30 min after EGF treatment of A431 cells. However, no change in Sp1 binding activity was found by EGF treatment. Moreover, the phosphatidylinositol 3 (PI3)-kinase inhibitor (wortmannin) and p38(MAPK) inhibitor (SB203580) inhibited the EGF-induced CREB phosphorylation and the expression of NF-IL6beta gene in cells. We also demonstrated that CREB was involved in regulating the NF-IL6beta gene transcriptional activity mediated by p38(MAPK). Our results suggested that PI3-kinase/p38(MAPK)/CREB pathway contributed to the EGF activation of NF-IL6beta gene expression.

Topics: Amino Acid Motifs; Androstadienes; Blotting, Western; CCAAT-Enhancer-Binding Protein-delta; Cell Line, Tumor; Cell Nucleus; Chromatin Immunoprecipitation; Cyclic AMP; Cyclic AMP Response Element-Binding Protein; Enzyme Inhibitors; Epidermal Growth Factor; Genes, Reporter; Humans; Imidazoles; Inflammation; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Plasmids; Promoter Regions, Genetic; Protein Binding; Pyridines; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Sp1 Transcription Factor; Time Factors; Transfection; Wortmannin

Endothelial membrane-bound thrombomodulin is a high affinity receptor for thrombin to inhibit coagulation. We previously demonstrated that the thrombin-thrombomodulin complex restrains cell proliferation mediated through protease-activated receptor (PAR)-1. We have now tested the hypothesis that thrombomodulin transduces a signal to activate the endothelial nitric-oxide synthase (NOS3) and to modulate G protein-coupled receptor signaling. Cultured human umbilical vein endothelial cells were stimulated with thrombin or a mutant of thrombin that binds to thrombomodulin and has no catalytic activity on PAR-1. Thrombin and its mutant dose dependently activated NO release at cell surface. Pretreatment with anti-thrombomodulin antibody suppressed NO response to the mutant and to low thrombin concentration and reduced by half response to high concentration. Thrombin receptor-activating peptide that only activates PAR-1 and high thrombin concentration induced marked biphasic Ca2+ signals with rapid phosphorylation of PLC(beta3) and NOS3 at both serine 1177 and threonine 495. The mutant thrombin evoked a Ca2+ spark and progressive phosphorylation of Src family kinases at tyrosine 416 and NOS3 only at threonine 495. It activated rapid phosphatidylinositol-3 kinase-dependent NO synthesis and phosphorylation of epidermal growth factor receptor and calmodulin kinase II. Complete epidermal growth factor receptor inhibition only partly reduced the activation of phospholipase Cgamma1 and NOS3. Prestimulation of thrombomodulin did not affect NO release but reduced Ca2+ responses to thrombin and histamine, suggesting cross-talks between thrombomodulin and G protein-coupled receptors. This is the first demonstration of an outside-in signal mediated by the cell surface thrombomodulin receptor to activate NOS3 through tyrosine kinase-dependent pathway. This signaling may contribute to thrombomodulin function in thrombosis, inflammation, and atherosclerosis.

Topics: Atherosclerosis; Blotting, Western; Calcium; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Calcium-Calmodulin-Dependent Protein Kinases; Catalysis; Cells, Cultured; Dose-Response Relationship, Drug; Endothelial Cells; Endothelium, Vascular; Epidermal Growth Factor; ErbB Receptors; Humans; Inflammation; Models, Biological; Mutation; Nitric Oxide; Nitric Oxide Synthase Type III; Peptides; Phosphatidylinositol 3-Kinases; Phosphorylation; Protein Structure, Tertiary; Receptor, PAR-1; Receptors, Thrombin; Reverse Transcriptase Polymerase Chain Reaction; Serine; Signal Transduction; src-Family Kinases; Threonine; Thrombin; Thrombomodulin; Time Factors; Tyrosine; Umbilical Veins

Inflammation is commonly associated with lung tumors. Since inflammatory mediators, including members of the interleukin-6 (IL-6) cytokine family, suppress proliferation of normal epithelial cells, we hypothesized that epithelial cells must develop mechanisms to evade this inhibition during the tumorigenesis. This study compared the cytokine responses of normal epithelial cells to that of premalignant cells.. Short-term primary cultures of epithelial cells were established from bronchial brushings. Paired sets of brushings were obtained from areas of normal bronchial epithelium and from areas of metaplastic or dysplastic epithelium, or areas of frank endobronchial carcinoma. In 43 paired cultures, the signalling through the signal transducer and activator of transcription (STAT) and extracellular regulated kinase (ERK) pathways and growth regulation by IL-6, leukemia inhibitory factor (LIF), oncostatin M (OSM), interferon-gamma (IFNgamma) or epidermal growth factor (EGF) were determined. Inducible expression and function of the leukemia inhibitory factor receptor was assessed by treatment with the histone deacetylase inhibitor depsipeptide.. Normal epithelial cells respond strongly to OSM, IFNgamma and EGF, and respond moderately to IL-6, and do not exhibit a detectable response to LIF. In preneoplastic cells, the aberrant signaling that was detected most frequently was an elevated activation of ERK, a reduced or increased IL-6 and EGF response, and an increased LIF response. Some of these changes in preneoplastic cell signaling approach those observed in established lung cancer cell lines. Epigenetic control of LIF receptor expression by histone acetylation can account for the gain of LIF responsiveness. OSM and macrophage-derived cytokines suppressed proliferation of normal epithelial cells, but reduced inhibition or even stimulated proliferation was noted for preneoplastic cells. These alterations likely contribute to the supporting effects that inflammation has on lung tumor progression.. This study indicates that during the earliest stage of premalignant transformation, a modified response to cytokines and EGF is evident. Some of the altered cytokine responses in primary premalignant cells are comparable to those seen in established lung cancer cell lines.

Topics: Cell Line, Transformed; Cell Line, Tumor; Cell Transformation, Neoplastic; Cells, Cultured; Cytokines; Densitometry; Epidermal Growth Factor; Epithelial Cells; Extracellular Signal-Regulated MAP Kinases; Fibroblasts; Histones; Humans; Inflammation; Interferon-gamma; Interleukin-6; Lung; Lung Neoplasms; Macrophages; Oncostatin M; Signal Transduction; Time Factors; Treatment Outcome

To investigate the effect of peroxisome proliferator-activated receptor gamma (PPAR-gamma) and its ligand, ciglitazone, on inflammatory regulation of human gallbladder epithelial cells (HGBECs) and to assess the effect of human epithelial growth factor (hEGF) on growth of HGBECs.. HGBECs were cultured in media containing hEGF or hEGF-free media. HGBECs were divided into normal control group, inflammatory control group and ciglitazone group (test group). Inflammatory control group and ciglitazone group were treated with 5 microg/L of human interleukin-1beta (hIL-1beta) to make inflammatory model of HGBECs. The ciglitazone group was treated with various concentrations of ciglitazone, a potent ligand of PPAR-gamma. Subsequently, interleukin-8 (IL-8), IL-6, and tumor necrosis factor-alpha (TNF-alpha) concentrations in all groups were measured. The data were analyzed statistically.. HGBECs were cultured in medium successfully. The longevity of HGBECs in groups containing hEGF was longer than that in hEGF-free groups. So was the number of HGBECs. The longest survival time of HGBEC was 25 d. The inflammatory model of HGBECs was obtained by treating with hIL-1beta. The concentrations of IL-6 and IL-8 in ciglitazone group were lower than those in inflammatory control group (P<0.05). The secretion of IL-6 in inflammatory control group was higher (350.31+/-37.05 microg/L) than that in normal control group (50.0+/-0.00 microg/L, P<0.001). Compared to normal control group, IL-8 concentration in inflammatory control was higher (P<0.05).. hEGF improves the growth of HGBECs in vitro. Ciglitazone inhibits the inflammation of HGBECs in vitro and has potential therapeutic effect on cholecystitis in vivo.

Topics: Cells, Cultured; Cholecystitis; Epidermal Growth Factor; Epithelial Cells; Gallbladder; Humans; Inflammation; Interleukin-6; Interleukin-8; Ligands; Models, Biological; PPAR gamma; Thiazolidinediones; Tumor Necrosis Factor-alpha

Many years ago, diphtheria toxin (DT) showed antitumor activity in mice and in humans, but it was unclear whether this depended on the toxicity of the molecule only or on its strong inflammatory-immunological property as well. To deal with this open question, we planned to treat a group of cancer patients with cross-reacting material 197 (CRM197). CRM197 is a nontoxic mutant of DT that shares the immunological properties of the native molecule and its ability to bind to heparin-binding epidermal growth factor (HB-EGF), the specific cell-membrane receptor for DT that is often overexpressed in cancer.. 25 outpatients with various advanced tumors who were refractory to standard therapies (23 subjects) or had refused, in whole or in part, conventional therapies (2 subjects) were treated with CRM197 injected subcutaneously in the abdominal wall, on alternate days, for 6 days. Three different dosages (1.7, 2.6, or 3.5 mg/day) were used according to the patient's degree of immunological reactivity to DT/CRM197 (none, moderate, or high).. After the first administration of CRM197, a significant increase in the number of circulating neutrophils and in the serum level of TNF-alpha was detected. Toxicities were minimal. Only patients with delayed-type hypersensitivity to DT/CRM197 had irritating skin reactions in the injection sites and a flu-like syndrome with fever. Pharmacokinetics showed a mean peak concentration (12.7 ng/ml) 12 h after the first injection and a mean half-life of 18.1 h. There were two complete and one partial responses (metastatic breast carcinoma, neuroblastoma, and metastatic breast carcinoma) lasting 4, 45+, and 15 months, respectively. Six cases of stable disease, lasting from 1 to 15 months, were also recorded.. CRM197 injected subcutaneously elicited an inflammatory-immunological reaction, caused tolerable toxicities, was absorbed to a good extent into the circulatory system, and exerted some degree of biological antitumor activity. A possible role of neutrophils and TNF-alpha in the mode of action of the molecule is hypothesized.

Topics: Aged; Aged, 80 and over; Bacterial Proteins; Cell Line, Tumor; Clinical Trials as Topic; Dose-Response Relationship, Drug; Epidermal Growth Factor; Female; Heparin; Humans; Inflammation; Male; Middle Aged; Neoplasms; Neutrophils; Time Factors; Treatment Outcome; Tumor Necrosis Factor-alpha

Epidermal growth factor (EGF) and its receptor (EGF-R) are attractive targets for cancer immunotherapy. Tolerance has been broken with an EGF-vaccine and antibodies against EGF have been produced in animals and in cancer patients. EGF also plays an important role in the inflammation stage of wound healing. Because this therapeutic approach may be of importance after surgery procedures in cancer patients, we decided to investigate the possible role of the EGF-vaccine in the croton-oil-induced ear edema and in the wound healing experimental animal models.. Mice were immunized with an EGF-vaccine by intramuscular injections and serum titers against EGF were measured through ELISA techniques. Control animals received saline.. Immunized mice produced antibodies against EGF while no antibody titers could be measured in control animals. Croton oil applied to the inner ear surface of EGF-vaccine treated mice caused a 61.3% lower ear punch weight and a 60.2% lower myeloperoxidase activity than control mice. In the EGF-vaccine treated animals, planimetry measurements and histological analysis did not led to significant impairment in tissue repair.. The EGF-vaccination in mice decreased the normal croton-oil-induced inflammation response, without apparent impairment in tissue healing.

Topics: Animals; Antibody Formation; Autoantibodies; Cancer Vaccines; Croton Oil; Dermatologic Agents; Drug Combinations; Ear Diseases; Edema; Epidermal Growth Factor; Image Processing, Computer-Assisted; Immunization; Inflammation; Injections, Intramuscular; Mice; Mice, Inbred BALB C; Skin; Wound Healing; Wounds, Penetrating

Many cells upon injury mount extensive, compensatory responses that increase cell survival; however, the intracellular signals that regulate these responses are not completely understood. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) has been implicated as a cytoprotective agent. We have previously demonstrated that pretreatment of human intestinal epithelial cells with HB-EGF significantly decreased cytokine-induced activation of inducible NO synthase mRNA expression and NO production and protected the cells from apoptosis and necrosis. However, the mechanisms by which HB-EGF exerts these effects are not known. Here we show that cytokine exposure (IL-1beta and IFN-gamma) induced NF-kappaB activation and IL-8 and NO production in DLD-1 cells. Transient expression of a dominant negative form of IkappaBalpha decreased NO production, suggesting that the cytokines stimulated NO production in part through activation of NF-kappaB. HB-EGF dramatically suppressed NF-kappaB activity and IL-8 release and decreased NO production in cells pretreated with HB-EGF. HB-EGF blocked NF-kappaB activation by inhibiting IkappaB kinase activation and IkappaB phosphorylation and degradation, thus interfering with NF-kappaB nuclear translocation, DNA-binding activity, and NF-kappaB-dependent transcriptional activity. The data demonstrate that HB-EGF decreases inflammatory cytokine and NO production by interfering with the NF-kappaB signaling pathway. Inhibition of NF-kappaB may represent one of the mechanisms by which HB-EGF exerts its potent anti-inflammatory and cytoprotective effects.

Topics: Active Transport, Cell Nucleus; Cell Line, Tumor; Cell Nucleus; Cytokines; DNA-Binding Proteins; Down-Regulation; Epidermal Growth Factor; Heparin; Heparin-binding EGF-like Growth Factor; HT29 Cells; Humans; I-kappa B Kinase; I-kappa B Proteins; Inflammation; Intercellular Signaling Peptides and Proteins; Interleukin-8; Intestinal Mucosa; NF-kappa B; NF-KappaB Inhibitor alpha; Nitric Oxide; Phosphorylation; Protein Serine-Threonine Kinases; Signal Transduction; Transcription, Genetic

The epidermal growth factor (EGF)-TM7 subgroup of G-protein-coupled receptors is composed predominantly of leukocyte-restricted glycoproteins defined by their unique hybrid structure, in which extracellular EGF-like domains are coupled to a seven-span transmembrane moiety via a mucin-like stalk. The EGF-TM7 group comprises mouse F4/80, human EGF module-containing mucin-like hormone receptor (EMR) 1, human EMR2, and human and mouse CD97, the genes for which map to human chromosome 19p13 and the syntenic regions of the mouse genome. In this study we describe the cloning and characterization of EMR3, a novel human EGF-TM7 molecule, and show the existence of its cellular ligand. The EMR3 gene maps closely to the existing members of the EGF-TM7 family on human chromosome 19p13.1 and, in common with other EGF-TM7 genes, is capable of generating different protein isoforms through alternative splicing. Two alternative splice forms have been isolated: one encoding a 652-amino acid cell surface protein consisting of two EGF-like domains, a mucin stalk, and a putative G-protein-coupled receptor domain and the other encoding a truncated soluble form containing only two EGF-like domains. As with other members of the EGF-TM7 family, EMR3 mRNA displays a predominantly leukocyte-restricted expression pattern, with highest levels in neutrophils, monocytes, and macrophages. Through the use of soluble EMR3 multivalent probes we have shown the presence of a ligand at the surface of monocyte-derived macrophages and activated human neutrophils. These interactions suggest a potential role for EMR3 in myeloid-myeloid interactions during immune and inflammatory responses.

Topics: Alternative Splicing; Amino Acid Sequence; Base Sequence; Biotinylation; Chromosome Mapping; Chromosomes, Human, Pair 19; Cloning, Molecular; DNA, Complementary; Epidermal Growth Factor; Humans; Inflammation; Leukocytes; Ligands; Macrophages; Models, Chemical; Molecular Sequence Data; Monocytes; Mucins; Neutrophils; Protein Binding; Protein Isoforms; Protein Structure, Tertiary; Receptors, G-Protein-Coupled; Receptors, Peptide; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Tissue Distribution

Sterile tissue injury or infection initiates a local inflammatory response that mobilizes a systemic acute phase reaction resulting in, among other things, the induction of genes encoding the acute phase plasma proteins (APPs). In all vertebrates, a common set of APPs is increased and exerts essential protective functions. Haptoglobin (HP), one of the major APPs, acts as a high-affinity hemoglobin-binding protein and antioxidant. Liver is the major site of HP synthesis; however, regulated, low level expression is also detected in other organs. Induction of the Hp gene is mediated by interleukin-6-type cytokines and is synergistically enhanced by glucocorticoids. Growth stimulation of hepatic cells in vivo or in vitro suppresses the Hp gene-inducing effects of inflammatory cytokines. Receptors for IL-6 cytokines mediate induction of the Hp gene by the transcription factors signal transducer and activator of transcription-3 (STAT3) and CAAT/enhancer binding protein beta (C/EBPbeta), but attenuate the stimulation through co-activated STAT5 and mitogen-activated protein kinases, ERK-1 and ERK-2. The specificity by which the related cytokines, IL-6, oncostatin M, and leukemia inhibitory factor, regulate Hp gene transcription is determined by the profile of the cytokine receptor subunits expressed on the target cells and the relative extents by which these receptors activate the intracellular signaling pathways. The current hypothesis is that HP exerts an anti-inflammatory activity and that by the degree with which HP attenuates the inflammatory process, including the production of IL-6 cytokines, it determines the level and duration of acute phase expression of the Hp gene.

Topics: Animals; Antioxidants; CCAAT-Enhancer-Binding Protein-beta; DNA-Binding Proteins; DNA, Complementary; Dose-Response Relationship, Drug; Epidermal Growth Factor; Gene Expression Regulation; Haptoglobins; Humans; Inflammation; Interleukin-6; Male; Mice; Mice, Inbred C57BL; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Models, Genetic; Oncostatin M; Peptides; Promoter Regions, Genetic; Rats; Receptors, Cytokine; STAT3 Transcription Factor; Time Factors; Trans-Activators; Transcription, Genetic

The CD44 glycoprotein is expressed in multiple isoforms on a variety of cell types where it functions as a receptor for hyaluronan-mediated motility. Recently, interest has centered on CD44 heparan sulfate proteoglycan (HSPG) isoforms because of their potential to sequester heparin-binding growth factors and chemokines. Expression of these isoforms on ectodermal cells has recently been shown to regulate limb morphogenesis via presentation of fibroblast growth factor (FGF) 4/FGF 8 while expression on tumor cells was shown to sequester hepatocyte growth factor and promote tumor dissemination. To date, however, CD44 HSPG expression in tissue macrophages and lymphocytes has not been adequately investigated, despite the fact these cells actively synthesize growth factors and chemokines and indirect evidence that monocyte CD44 sequesters macrophage inflammatory protein-1beta. Here we show primary human monocytes rather than lymphocytes express CD44 HSPGs, but only following in vitro differentiation to macrophages or activation with the proinflammatory cytokine interleukin-1alpha or bacterial lipopolysaccharide. Furthermore, we show these isoforms are preferentially modified with heparan rather than chondroitin sulfate, bind the macrophage-derived growth factors FGF-2, vascular endothelial growth factor, and heparin-binding epidermal growth factor with varying affinities (K(d) 25-330 nM) and in the case of FGF-2, can stimulate productive binding to the high affinity tyrosine kinase FGF receptor 1 (FGFR1). In contrast, we find no evidence for significant binding to C-C chemokines. Last, we confirm by immunofluorescent antibody staining that inflamed synovial membrane macrophages express CD44 HSPGs and that expression is greatest in cells containing high FGF-2 levels. These results suggest a paracrine role for macrophage CD44 HSPG isoforms in the regulation of growth factor action during inflammation.

Topics: Cell Differentiation; Chemokines; Endothelial Growth Factors; Epidermal Growth Factor; Fibroblast Growth Factor 2; Heparan Sulfate Proteoglycans; Heparin-binding EGF-like Growth Factor; Humans; Hyaluronan Receptors; Inflammation; Inflammation Mediators; Intercellular Signaling Peptides and Proteins; Interleukin-1; Lymphokines; Macrophages; Monocytes; Paracrine Communication; Protein Binding; Protein Isoforms; Receptor Protein-Tyrosine Kinases; Receptor, Fibroblast Growth Factor, Type 1; Receptors, Fibroblast Growth Factor; Synovial Membrane; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

Epidermal keratinocytes respond to injury by becoming activated, i.e. hyperproliferative, migratory, and proinflammatory. These processes are regulated by growth factors and cytokines. One of the markers of activated keratinocytes is keratin K6. We used a novel organ culture system to show that tumor necrosis factor alpha (TNFalpha) induces the expression of K6 protein and mRNA in human skin. Multiple isoforms of K6 are encoded by distinct genes and have distinct patterns of expression. By having shown previously that proliferative signals, such as epidermal growth factor (EGF), induce expression of the cytoskeletal protein keratin K6b, we here demonstrate that the same isoform, K6b, is also induced by TNFalpha, a proinflammatory cytokine. Specifically, TNFalpha induces the transcription of the K6b gene promoter. By using co-transfection, specific inhibitors, and antisense oligonucleotides, we have identified NFkappaB and C/EBPbeta as the transcription factors that convey the TNFalpha signal. Both transcription factors are necessary for the induction of K6b by TNFalpha and act as a complex, although only C/EBPbeta binds the K6b promoter DNA. By using transfection, site-directed mutagenesis, and footprinting, we have mapped the site that responds to TNFalpha, NFkappaB, and C/EBPbeta. This site is separate from the one responsive to EGF and AP1. Our results show that the proinflammatory (TNFalpha) and the proliferative (EGF) signals in epidermis separately and independently regulate the expression of the same K6b keratin isoform. Thus, the cytoskeletal responses in epidermal cells can be precisely tuned by separate proliferative and inflammatory signals to fit the nature of the injuries that caused them.

Topics: Base Sequence; Binding Sites; CCAAT-Enhancer-Binding Protein-beta; Cell Division; DNA Footprinting; Epidermal Cells; Epidermal Growth Factor; Epidermis; Fluorescent Antibody Technique; HeLa Cells; Humans; Inflammation; Keratinocytes; Keratins; Molecular Sequence Data; Mutation; NF-kappa B; Oligonucleotides, Antisense; Promoter Regions, Genetic; Protein Isoforms; Response Elements; Transcription Factor AP-1; Transcriptional Activation; Transfection; Tumor Necrosis Factor-alpha

Adhesion molecules are involved in inflammatory and repair processes of the bronchial epithelium. ICAM-1 is mainly involved in inflammatory reactions, whereas integrins, such as alpha3beta1, are mainly involved in repair processes.. Using bronchial biopsies from 10 asthmatics and eight controls, we first evaluated by immunohistochemistry expression of alpha3beta1 and ICAM-1 in intact and damaged epithelium. Then, using the human pulmonary epithelial cell line WI-26 VA, we studied, by flow-cytometry, the modulation of ICAM-1 and alpha3beta1 expression, and, by ELISA, the release of fibronectin by proinflammatory cytokines, such as IL-5, and anti-inflammatory cytokines, such as IL-4, TGF-beta, and EGF.. alpha3beta1 expression was slightly higher in asthma than in controls, as well as in damaged epithelium than in undamaged epithelium. ICAM-1 expression was higher in asthma than in controls, and similarly distributed in intact or damaged epithelium. In vitro, alpha3beta1 was significantly increased by TGF-beta, EGF, and IL-4, and significantly decreased by IL-5. Fibronectin release was significantly increased by TGF-beta and IL-4, unchanged by EGF, and slightly but significantly decreased by IL-5. ICAM-1 expression was significantly decreased by TGF-beta and IL-4, unchanged by EGF, and significantly increased by IL-5.. These differences in adhesion molecule expression and fibronectin release may be important in epithelial cell inflammation and repair.

Topics: Adolescent; Adult; Aged; Asthma; Biopsy; Bronchi; Cell Line; Cytokines; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Epithelial Cells; Fibronectins; Flow Cytometry; Humans; Inflammation; Integrin alpha3beta1; Integrins; Intercellular Adhesion Molecule-1; Interleukin-4; Middle Aged; Respiratory Mucosa; Transforming Growth Factor beta

Placental inflammations (villitis) are accompanied by loss of the syncytiotrophoblast, which is the cellular barrier separating maternal blood from fetal tissue in the villous placenta. We propose that syncytiotrophoblast loss is mediated by adhesion of activated maternal monocytes. This hypothesis was tested with a co-culture model of peripheral blood monocytes and placental syncytiotrophoblasts. We find that LPS-activated monocytes adhere to interferon-gamma (IFN-gamma)-treated syncytiotrophoblasts via monocyte LFA-1 for >48 h, during which time the monocytes induce trophoblast apoptosis and subsequent damage of the trophoblast layer. Optimal monocyte-mediated syncytiotrophoblast death requires both lipopolysaccharide (LPS) and IFN-gamma and is inhibited by either anti-tumor necrosis factor (TNF) antibody or epidermal growth factor. Syncytiotrophoblast damage is largely limited to culture surfaces in the vicinity of bound monocytes. These results show that activated maternal monocytes bound to the placental barrier can induce focal damage mediated by the inflammatory cytokine TNF-alpha and suggest a route for maternal leukocyte infiltration into the fetal stroma.

Topics: Antibodies, Monoclonal; Apoptosis; Cell Adhesion; Chorionic Villi; Epidermal Growth Factor; Female; Giant Cells; Humans; Inflammation; Interferon-gamma; Lipopolysaccharides; Lymphocyte Function-Associated Antigen-1; Monocytes; Pregnancy; Stromal Cells; Trophoblasts; Tumor Necrosis Factor-alpha

The epitope of mouse monoclonal antibody (mAb) EP-5C7, which binds to and blocks both human E- and P-selectin, was mapped onto the protein structure of E-selectin. Analyses with E- and L-selectin chimeric proteins and randomly mutagenized E-selectins demonstrated that the EP-5C7 epitope consists of the amino acid residues at positions 21, 22, 23, 119 and 120 of E-selectin. The binding of three neutralizing anti-E-selectin mAb's (E-1E4, H18/7 and CL2), whose epitopes were found to overlap with the E-selectin binding site for carbohydrate ligands, was not affected by the amino acid substitutions at these five positions. Inspection of the three-dimensional structure of E-selectin indicated that the EP-5C7 epitope is located near the junction between the lectin and EGF-like domains. The ligand binding site was distant from the EP-5C7 epitope, suggesting that the amino acid residues in the EP-5C7 epitope play an important role other than ligand binding in selectin-mediated cell adhesion.

Topics: Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Murine-Derived; Binding Sites; E-Selectin; Epidermal Growth Factor; Epitope Mapping; Epitopes; Humans; Inflammation; Lectins; Mice; Mutagenesis; Neutralization Tests; P-Selectin; Recombinant Fusion Proteins

An increase in myofibroblast number may be necessary for wound healing but may also lead to postinflammatory scarring. We have, therefore, studied the role of mediators important in inflammatory bowel disease in regulating proliferation of human colonic myofibroblasts. Using primary cultures of these cells, we have shown increases in [3H]thymidine incorporation in response to platelet-derived growth factor (EC50 = 14 ng/ml), basic fibroblast growth factor (EC50 = 2.2 ng/ml), and epidermal growth factor (EC50 = 1.1 ng/ml). Coulter counting of cell suspensions demonstrated increases in cell number with these growth factors along with insulin-like growth factor-I and -II. In addition the proinflammatory cytokines IL-1beta and TNF-alpha produced increases in [3H]thymidine incorporation. IL-1beta and platelet-derived growth factor together produced an increase in [3H]thymidine greater than either agonist alone; this effect was not, however, seen when we examined changes in cell numbers. Finally, we demonstrate a mechanism whereby these responses may be downregulated: vasoactive intestinal peptide (1 microM) elevates cyclic AwMP in these cells 4. 2-fold over control and produces a dose-related inhibition of platelet-derived growth factor-driven proliferation with a maximum inhibition of 33% at 1 microM.

Topics: Cell Division; Cells, Cultured; Colon; Epidermal Growth Factor; Fibroblast Growth Factor 2; Fibroblasts; Humans; Inflammation; Inflammation Mediators; Insulin-Like Growth Factor I; Insulin-Like Growth Factor II; Interleukin-1; Intestinal Mucosa; Middle Aged; Platelet-Derived Growth Factor; Tumor Necrosis Factor-alpha

Growth factors are important substances in the central control of wound healing during the exudative phase. Although these peptides have been applied frequently to chronic wounds in clinical studies, little is known about the naturally occurring levels at the wound site in correlation to healing in superficial wounds. We have therefore investigated the presence of these cytokines in partial thickness wounds. In 16 patients undergoing reconstructive surgery, split-thickness skin wounds were enclosed in cutaneous vinyl chambers filled with 2.5 ml of saline. Chambers placed over unwounded skin served as controls. After 24 hours, the accumulated wound fluid was harvested and replaced by 2.5 ml of saline until the wounds were healed. Wound fluid was centrifuged, aliquoted, and frozen at -70 degrees C. Samples were analyzed for protein and growth factors (insulin-like growth factor-1, epidermal growth factor, basic fibroblast growth factor, platelet-derived growth factor-AB, interleukin-1alpha, and transforming growth factor-beta1 and -beta2) and insulin-like growth factor-binding proteins 1 and 3 using a monoclonal Sandwich enzyme-linked immunosorbent assay and radioimmunoassay. All wounds healed in the liquid environment within 7 days (macroscopically) and 11 days (barrier function), respectively. In wound fluid, protein concentrations dropped from 5 mg/ml on day 1 to a baseline of 0.1 mg (unwounded skin), indicating a return of the barrier function. All growth factors could be measured already after 24 hours postwounding. However, the concentrations measured varied from 10 to more than 10,000 pg/ml between the different factors. The highest range was found for insulin-like growth factor-1 (21,000 to 41,000 pg/ml), the lowest for epidermal growth factor (3 to 63 and 3 to 88 pg/ml, respectively). Two different patterns of kinetics were distinguished: (1) a high initial peak decreasing to baseline values or below serum levels by the time of healing (insulin-like growth factor-1, insulin-like growth factor binding protein-1, -3, basic fibroblast growth factor, epidermal growth factor, platelet-derived growth factor-AB, transforming growth factor-beta1) and (2) a low initial concentration followed by an increase to a maximum at the time of epithelialization (interleukin-1alpha, transforming growth factor-beta2). Comparing the growth factor levels measured to serum baseline values, it was found that four of the growth factors appeared in wound fluid at above serum

Topics: Adult; Cytokines; Dermatologic Surgical Procedures; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Epithelium; Exudates and Transudates; Female; Fibroblast Growth Factor 2; Growth Substances; Humans; Inflammation; Insulin-Like Growth Factor Binding Protein 1; Insulin-Like Growth Factor Binding Protein 3; Insulin-Like Growth Factor I; Interleukin-1; Male; Middle Aged; Plastic Surgery Procedures; Platelet-Derived Growth Factor; Proteins; Skin; Time Factors; Transforming Growth Factor beta; Wound Healing

Both oxidized low density lipoprotein (ox-LDL) and platelet-derived growth factor (PDGF) have been implicated in the genesis of various inflammatory responses, including atherosclerosis. We demonstrate here a novel interaction between specific oxidized lipids derived from ox-LDL and PDGF. The lipid moieties of ox-LDL caused concentration-dependent inactivation of PDGF as measured by loss of its mitogenic activity and its binding to high affinity receptors. Reverse-phase and normal-phase HPLC were used to purify the inactivating component in the lipid mixture. By fast atom bombardment mass spectrometry and infrared spectroscopy, we identified the inactivating lipids as the 9- and 13-hydroperoxy derivatives of cholesteryl linoleate, cholesteryl hydroperoxyoctadecadienoate. When a series of cholesteryl esters were subjected to oxidizing conditions, only those containing two or more double bonds caused inactivation of PDGF; the extent of inactivation increased with increased levels of oxidation. Exposing PDGF to cumene hydroperoxide, t-butyl hydroperoxide, or hydrogen peroxide did not affect the activity of the mitogen. The oxidized lipid had no effect on the mitogenic activity of epidermal growth factor but did abolish the mitogenic activity of basic fibroblast growth factor and the antiproliferative activity of transforming growth factor beta1. The inactivation of PDGF and other cytokines by lipid hydroperoxides may occur in such processes as vascular disease, inflammation, and wound healing.

Topics: Arteriosclerosis; Cholesterol Esters; Epidermal Growth Factor; Fibroblast Growth Factor 2; Humans; Inflammation; Linoleic Acids; Lipid Peroxides; Lipoproteins, LDL; Mass Spectrometry; Peroxides; Platelet-Derived Growth Factor; Spectrum Analysis

To investigate the effects of recombinant human epidermal growth factor (rhEGF) eye drops on corneal wound healing.. Twenty-four white rabbits were randomly divided into 4 groups, 6 rabbits 12 eyes each. Anterior keratectomy of 8 mm in diameter and 1/3 cornea in thickness was performed on each eye. Each of the following concentrations of rhEGF: 1, 10, 100 microg/ml eye drops or normal saline (control) was applied four times daily for a week respectively for one group. The wound area was determined by computer imaging analysis.. The mean epithelial healing rate of rhEGF 1, 10, 100 microg/ml groups was 9.31, 9.96, 9.31 mm(2)/day respectively, significantly greater than 8.11 mm(2)/day of the control group. The action of rhEGF of 10 microg/ml was somewhat better than that of 1 or 100 microg/ml, and no significant difference was noticed among the three rhEGF groups. Moderate inflammation and corneal neovascularization were induced in the rhEGF 100 microg/ml treated group.. rhEGF 1 - 10 microg/ml can accelerate corneal wound healing in the rabbit with no adverse side-effects. It may be used to treat serious corneal trauma and ulcer clinically.

Topics: Animals; Cornea; Corneal Injuries; Epidermal Growth Factor; Female; Inflammation; Male; Rabbits; Recombinant Proteins; Wound Healing

Since many cytokines have been identified in chronically inflamed human synovium, it is possible that particular cytokines or combinations of cytokines play dominant roles in driving or inhibiting metabolic processes important to inflammation. To assess these possibilities, we compared selected effects of individual cytokines and their binary, ternary, and higher combinations in human synovial cell cultures.. Cytokines studied known to occur in human synovial tissue included: interleukin 1beta (IL-1beta), IL-6, tumor necrosis factor-alpha, granulocyte macrophage colony stimulating factor, interferon-gamma, acidic fibroblast growth factor (aFGF), basic FGF (bFGF), platelet derived growth factor, transforming growth factor-beta1, connecting tissue activating peptide-III, and epidermal growth factor. The growth related effects of these agents singly and in combinations were assessed by measuring newly synthesized [3H]DNA and [14C]GAG (glycosaminoglycan) in human synovial cell cultures. Cytokine induced synthesis of prostaglandin E2 (PGE2) was measured by ELISA.. Most cytokine combinations resulted in additive/synergistic anabolic effects, except when IL-1beta was present; IL-1beta was markedly antagonistic to the mitogenic effects of other cytokines tested. Combinations of platelet derived cytokines were the most potent stimulators of DNA synthesis, while combinations of synovial derived cytokines were more active in stimulating GAG synthesis. Synovial cells exposed simultaneously to both platelet and synovial derived cytokines produced large quantities of [14C]GAG and showed a modest increase in [3H]DNA synthesis. IL-1beta, alone or in combinations, was dominant with respect to stimulation of PGE2 synthesis. Acetylsalicylic acid substantially interfered with all the effects of cytokine combinations measured.. Quantitative alterations in synovial cell synthesis of GAG and DNA varied greatly depending on the ambient mixture of cytokines. Virtually all combinations of cytokines tested gave rise to large increases in synovial cell synthesis of GAG. Four platelet derived cytokines, a "physiologic combination," appeared to be dominant agents in stimulating DNA synthesis. This effect was profoundly reduced by the antagonistic effect of IL-1beta, mediated in part by PGE2. The patterns of cytokine combination induced metabolic effects suggest that the "cytokine network" has a significant measure of redundancy with respect to control of synovial cell metabolism.

Topics: Arthritis, Rheumatoid; Aspirin; Cells, Cultured; Connective Tissue; Culture Media, Conditioned; Cytokines; Dinoprostone; DNA; Drug Synergism; Epidermal Growth Factor; Fibroblast Growth Factors; Glycosaminoglycans; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Hydrocortisone; Inflammation; Insulin-Like Growth Factor I; Interferon-gamma; Interleukin-6; Osteoarthritis; Peptides; Platelet-Derived Growth Factor; Recombinant Proteins; Synovial Membrane; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Topics: Animals; Base Sequence; Cytokines; DNA Primers; Epidermal Growth Factor; Gene Expression; Granulocyte-Macrophage Colony-Stimulating Factor; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Inflammation; Interferon-gamma; Interleukin-10; Ischemia; Kidney; Major Histocompatibility Complex; Male; Mice; Mice, Inbred BALB C; Mice, Knockout; Molecular Sequence Data; Reference Values; RNA, Messenger; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

The oviduct provides the environment in which fertilization of the egg and subsequent development of the preimplantation mouse embryo occurs, but little is known about the oviduct's capacity to produce growth factors or cytokines that may influence these preimplantation events. Northern blot analysis and/or immunohistochemistry were employed to examine the expression or cellular distribution, respectively, of the growth factors heparin-binding epidermal-like growth factor (HB-EGF), transforming growth factor (TGF) alpha, epidermal growth factor (EGF), insulin-like growth factor-I (IGF-I), TGF beta 1, TGF beta 2, and TGF beta 3; of estrogen-regulated lactoferrin (LF); and of the cytokines interleukin (IL)-1 alpha and IL-1 beta in the mouse oviduct during the preimplantation period (Days 1-4 [Day 1 = vaginal plug]) and 7 days after ovariectomy. The results demonstrated that, except for EGF, each of the growth factors and the LF genes are expressed in the ampulla and isthmus regions of the oviduct throughout the preimplantation period. Prominent immunostaining in secretory epithelial cells was noted for HB-EGF, TGF alpha, IGF-I, TGF beta 1, and TGF beta 2, and LF. Less intense immunostaining in the serosa and/or smooth muscle was also noted for TGF alpha, IGF-I, and TGF beta 1. In contrast, intense immunostaining in smooth muscle was noted for TGF beta 2, and TGF beta 3 was detected exclusively in smooth muscle cells. The abundance of these mRNAs was relatively constant during the preimplantation period, and ovariectomy did not reduce the levels of these mRNAs. In contrast to these growth factors, the cytokine mRNAs examined (IL-1 alpha and IL-1 beta) were at or below the limits of detection under these experimental conditions, and inflammatory leukocytes (LF-immunopositive neutrophils, IL-1 beta-immunopositive monocytes/macrophages, or peroxidase-positive eosinophils) were not detected in the oviduct, but were abundant in the adjacent uterine stroma on Day 1. These studies show that several growth factors are synthesized by the mouse oviduct and suggest that ovarian steroids do not play a major role in modulating expression of these genes in the oviduct during the preimplantation period. Furthermore, unlike the uterus on Day 1, the oviduct does not exhibit an inflammatory response to mating.

Topics: Animals; Blotting, Northern; Embryonic Development; Eosinophils; Epidermal Growth Factor; Fallopian Tubes; Female; Gene Expression; Growth Substances; Inflammation; Insulin-Like Growth Factor I; Interleukin-1; Lactoferrin; Leukocytes; Mice; Monocytes; Neutrophils; Ovariectomy; Pregnancy; RNA, Messenger; Transforming Growth Factor alpha; Transforming Growth Factor beta

Cytokines have profound effects on various aspects of granulomatous tissue formation. However, there is little information regarding their distribution during tissue development. This study investigated the temporal and spatial distribution of transforming growth factor-beta (TGF-beta), platelet-derived growth factor (PDGF), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1) and IL-1 beta in developing granulomatous tissue.. Murine chronic granulomatous air pouches were induced and full thickness biopsies taken at intervals up to 28 days. Samples were prepared for immunohistochemistry and labeled using antibodies against TGF-beta, bFGF, PDGF, EGF, TNF-alpha, IL-1 alpha and IL-1 beta.. Immunoreactivity to TGF-beta, PDGF, TNF-alpha, IL-1 alpha and IL-1 beta was localized to a proportion of macrophages within the granulomatous tissue. Immunopositive macrophage numbers increased with time, and with the exception of PDGF were associated with areas of fibrogenesis between days 14 to 28. Heterogeneous labeling of capillaries for EGF was observed within the granulomatous tissue juxtaposed to dermal musculature. Diffuse labeling of bFGF, associated with extracellular matrix, was always observed. After day 14, bFGF immunoreactivity was discretely localized to endothelial cells and the basement membrane of vessels within the granulomatous tissue. TGF-beta immunoreactivity was also associated with extracellular matrix components, being most intense in the area of fibrogenesis between 14 and 28 days. Occasional fibroblasts were also labeled with TGF-beta in this region.. The spatial and temporal confinement of the individual cytokines suggests that a sequential coordinated process of repair and fibrosis is occurring. It is hoped that these observations will provide a more effective therapeutic approach for the sequential application of cytokines in abnormalities of wound healing.

Topics: Animals; Croton Oil; Cytokines; Epidermal Growth Factor; Female; Fibroblast Growth Factor 2; Granuloma; Immunohistochemistry; Inflammation; Interleukin-1; Mice; Mice, Inbred Strains; Platelet-Derived Growth Factor; Skin; Skin Diseases; Time Factors; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Wound Healing

In the skin, wounding initiates a complex array of physiological processes mediated by growth factors and inflammatory mediators which stimulate tissue repair and protect against infection. We report that primary cultures of human keratinocytes and a mouse keratinocyte cell line respond to the inflammatory stimuli gamma-interferon and lipopolysaccharide or tumor necrosis factor-alpha by producing nitric oxide and hydrogen peroxide, two reactive mediators that are important in nonspecific host defense. Nitric oxide is produced by the l-arginine- and NADPH-dependent enzyme, nitric oxide synthase. In murine keratinocytes, optimal enzymatic activity was found to be dependent on Ca2+ and calmodulin as well as on glutathione. Inflammatory mediators were also found to inhibit the growth of keratinocytes, an effect that could be reversed by a nitric oxide synthase inhibitor. Epidermal growth factor (EGF), which promotes wound healing by stimulating cellular proliferation, was found to be a potent antagonist of reactive nitrogen and reactive oxygen intermediate production by keratinocytes. EGF also reversed the growth inhibitory actions of the inflammatory mediators. These data suggest that nitric oxide produced by keratinocytes is important in the control of cellular proliferation during wound healing. Our findings that EGF effectively regulates the production of free radicals by keratinocytes may represent an important pathway by which this growth factor not only stimulates epidermal cell proliferation but also facilitates the resolution of inflammation following wounding.

Topics: Amino Acid Oxidoreductases; Animals; Cell Division; Cells, Cultured; Epidermal Growth Factor; Humans; Hydrogen Peroxide; Inflammation; Keratinocytes; Mice; Nitric Oxide; Nitric Oxide Synthase; Wound Healing

Recent evidence has implicated cytokines and growth factors in the initiation of parturition in women. In the present study, the amnion-derived cell line WISH was used to determine whether proinflammatory cytokines (interleukins 1 beta, 6, and 8, tumor necrosis factor-alpha, and granulocyte/macrophage colony stimulating factor) could amplify epidermal growth factor-induced prostaglandin E2 production. WISH cells were preincubated with cytokines (0.0001-10 ng/ml) for 60 min and then challenged with EGF (10 ng/ml) for 4 hrs after which PGE2 production was measured by radioimmunoassay. EGF, IL-1 beta and TNF-alpha alone caused a dose-dependent increase in PGE2 production, while IL-6, IL-8 and GM-CSF were ineffective over the dose range tested. When cells were preincubated with IL-1 beta or TNF-alpha, there was a dose-dependent potentiation of EGF-induced PGE2 production that was greater than the sum of EGF alone and IL-1 beta or TNF-alpha alone. In each case, the minimum dose of IL-1 beta or TNF-alpha which amplified EGF-induced PGE2 production was 0.1 ng/ml (p less than 0.05, Student's t-test). These data show that low concentrations of IL-1 beta or TNF-alpha may serve to amplify EGF-mediated PGE2 biosynthesis in amnion-derived cells and suggest that cytokines may modulate EGF function in responsive cells.

Topics: Amnion; Cell Line; Cytokines; Dinoprostone; Drug Synergism; Epidermal Growth Factor; Female; Humans; Inflammation; Pregnancy; Radioimmunoassay

Previous studies have shown that bilateral decentralization of the superior cervical ganglia (SCG; decentralization) attenuates allergen-induced pulmonary inflammatory responses in male rats sensitized to the nematode Nippostrongylus brasiliensis. The present report examines the neuronal and glandular mechanisms mediating the protection against pulmonary inflammation afforded by decentralization. Tissues and organs innervated by the SCG are responsible for this protection since, in a manner similar to decentralization, bilateral removal of the SCG (ganglionectomy) reduced anaphylaxis-induced accumulation of inflammatory cells in bronchoalveolar lavage fluid. Removal of the submandibular gland (sialadenectomy) did not modify the severity of the pulmonary inflammation, but concurrent sialadenectomy and decentralization abolished the protective effect of decentralization. Thus, we postulate that cervical sympathetic nerves tonically inhibit release of anti-inflammatory factors from submandibular glands. No relationship was found between noradrenaline and serotonin content of submandibular glands and the degree of protection against pulmonary inflammation offered by decentralization and ganglionectomy. Both decentralization and ganglionectomy appeared to increase the level of transcripts that encode immunomodulatory growth factors (nerve growth factor and epidermal growth factor) in submandibular glands, but these denervations evidently did not modify the transcripts for TGF beta 2. Systemic inflammatory events are regulated by the central nervous system at a level superior to the SCG probably through modulation of immunoregulatory factors in submandibular glands.

Topics: Anaphylaxis; Animals; Bronchoalveolar Lavage Fluid; Catecholamines; Epidermal Growth Factor; Ganglia, Sympathetic; Immunization; Inflammation; Lung; Male; Nerve Growth Factors; Nippostrongylus; Polymerase Chain Reaction; Rats; Rats, Inbred Strains; Submandibular Gland; Sympathectomy; Transforming Growth Factor beta

Transforming growth factor-beta (TGF-beta) has been implicated as having a role in inflammatory responses by inducing cellular infiltration and the release of inflammatory cytokines. In this study, the IEC-6 rat intestinal epithelial cell line was used as a model to assess the effect of TGF-beta 1 on the expression of various plasma membrane determinants. TGF-beta 1 induced a dose-dependent increase in the percentage of cells expressing surface secretory component (SC) and class I major histocompatibility (MHC) antigens. However, the expression of class II MHC was unaffected. In contrast, epidermal growth factor had no effect on any of the surface proteins studied. The TGF-beta 1-enhanced expression of SC was accompanied by an enhanced binding of polymeric, but not monomeric, immunoglobulin A (IgA). Preincubation of the TGF-beta 1-treated cells with an anti-human beta-galactosyltransferase (beta-GT) antiserum did not block the binding of the anti-SC antibody, indicating that the TGF-beta-induced increase in SC staining was due to SC expression and not the polymeric immunoglobulin-binding enzyme, beta-GT. These results indicate that TGF-beta 1 may be important in immune functions involving intestinal epithelial cells by enhancing the expression of surface class I MHC antigens and SC, a protein responsible for the transport of polymeric IgA into the intestinal lumen.

Topics: Animals; Cell Division; Cell Line; Dose-Response Relationship, Drug; Epidermal Growth Factor; Epithelium; Galactosyltransferases; Gene Expression Regulation; Histocompatibility Antigens Class I; Immunoglobulin A; Inflammation; Intestines; Membrane Proteins; Rats; Secretory Component; Transforming Growth Factor beta

Conditioned media obtained from fibroblasts cultured from rheumatoid and certain other inflammatory synovia were observed to stimulate [3H]thymidine incorporation in an indicator murine fibroblast line. Synovial fibroblasts derived from the joints of patients with osteoarthritis did not display this property. This effect persisted in culture for many weeks and occurred in the absence of co-stimulatory immune cells. Antibody neutralization studies implicated a role for basic fibroblast growth factor (bFGF), transforming growth factor beta (TGF-beta), granulocyte/macrophage colony-stimulating factor (GM-CSF), and interleukin 1 beta (IL-1 beta) in the increased proliferative activity of synovial fibroblast-conditioned media. Synovial cell synthesis of bFGF, TGF beta 1, GM-CSF, IL-1 beta, and IL-6 was confirmed by 35S-methionine labeling and immunoprecipitation. The constitutive production of inflammatory and mitogenic cytokines by synovial fibroblasts may represent the result of long-term, phenotypic changes that occurred in vivo. Persistent cytokine production by synovial fibroblasts may play an important role in the continued recruitment and activation of inflammatory cells in chronic arthritis and in the formation of rheumatoid pannus.

Topics: Animals; Arthritis, Rheumatoid; Cell Division; Cell Line; Cells, Cultured; Cytokines; DNA Replication; Epidermal Growth Factor; Fibroblast Growth Factor 2; Fibroblasts; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Inflammation; Interleukin-1; Interleukin-6; Mice; Osteoarthritis; Synovial Membrane; Transforming Growth Factor beta

Human peripheral blood leukocytes can stimulate G1(G0)-arrested mouse skin keratinocytes to enter the cell cycle again and synthesize DNA at the maximum rate 15-20 hr later. This growth-promoting activity is released by the monocyte fraction and is shown to have characteristics that have been reported for interleukin 1 (IL-1). Pure IL-1 is active in stimulating keratinocyte cultures as was shown with recombinant human IL-1. An IL-1-like protein released by monocytes-macrophages could explain the hyperproliferative epidermis found in certain types of inflammatory skin diseases.

Topics: Animals; Cell Division; Cells, Cultured; Concanavalin A; DNA Replication; Epidermal Cells; Epidermal Growth Factor; Fibroblasts; Humans; Inflammation; Interleukin-1; Interphase; Leukocytes; Lymphocytes; Mice; Mice, Inbred C3H; Muscles; Skin; Skin Diseases

Topics: Angiogenesis Inducing Agents; Animals; Antigens; Biocompatible Materials; Blood Platelets; Burns; Cicatrix; Collagen; Epidermal Growth Factor; Fibroblast Growth Factors; Fluorescent Antibody Technique; Granulocytes; Growth Substances; Humans; Hypertrophy; Inflammation; Laminin; Leukocytes; Macrophages; Pulmonary Fibrosis; Rabbits; Silicone Elastomers; Wound Healing; Wound Infection