ovalbumin and carvacrol

Asthma is a complex inflammatory disease which affects multiple individuals worldwide especially pediatric ages.. This study aimed to assess the possible protective effect of carvacrol, as natural antioxidant anti-inflammatory drug, against bronchial asthma induced experimentally in rats.. Rats were randomly allocated into 5 groups; a normal control group, control drug group received only carvacrol, an asthma control group, a standard treatment group receiving dexamethasone (DEXA) and carvacrol treatment group. Bronchial asthma was induced by sensitization with i.p dose followed by challenge with intranasal dose of ovalbumin (OVA). 24 h after the last challenge, absolute eosinophil count (AEC) were determined in bronchoalveolar lavage fluids (BALF). Immunoglobulin E (IgE) was determined in serum. Inflammatory biomarkers like Interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin 13 (IL-13), tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) were also measured in BALF. Nitrosative stress biomarker namely inducible nitric oxide synthase (iNOS) was determined in BALF as well as oxidative stress biomarkers namely superoxide dismutase (SOD), glutathione (GSH) and malondialdehyde (MDA) were determined in lung tissue. Additionally, histopathological study, immunohistochemical study of UCN and western blot analysis of SP-D were performed.. Carvacrol administration significantly reduced the values of AEC, IgE, IL-4, IL-5, IL-13, TNF-α, IFN-γ, iNOS and MDA, while it significantly increased the values of SOD and GSH as compared to the asthmatic group. Histopathological, immunohistochemical and western blot study reinforced the biochemical results.. Carvacrol may be a promising protective agent against bronchial asthma induced experimentally in rats.

Topics: Animals; Anti-Asthmatic Agents; Anti-Inflammatory Agents; Asthma; Blotting, Western; Cymenes; Disease Models, Animal; Immunologic Factors; Interleukin-13; Interleukin-4; Interleukin-5; Male; Nitric Oxide Synthase Type II; Ovalbumin; Pulmonary Surfactant-Associated Protein D; Rats; Rats, Wistar; Real-Time Polymerase Chain Reaction

The applications of carvacrol are limited due to its poor stability and water solubility, and high volatility; however, ovalbumin can be used to encapsulate hydrophobic molecules, improve their aqueous solubility, and reduce their volatility. In this study, we fabricated ovalbumin-carvacrol nanoparticles (OCGns) under different pH (2, 5, 7, and 9) conditions using a gel embedding method and investigated their physicochemical and antibacterial properties. Rheological experiments revealed that the G' of ovalbumin gels (OGs) prepared under different pH conditions were OG-2 > OG-7 > OG-9 > OG-5. Carvacrol addition reduced the tight structure of ovalbumin and carvacrol under pH 5 and 7 conditions, with hardness first decreasing and then increasing, but increasing under pH 2 and 9 conditions. Fluorescence and infrared spectroscopy indicated complex formation, with carvacrol increasing the average diameter of nanoparticles prepared at pH 2, 5, 7, and 9. Encapsulation reached 89.34 and 91.86% at pH 2 and 9, respectively; however, inhibition experiments revealed that the minimum inhibitory concentration of OCGn-2 against Gram-positive Bacillus cereus and Salmonella (0.08 and 0.16 mg mL-1, respectively) was lower than that of OCGn-9 (both 0.28 mg mL-1). Moreover, OCGn-2 possessed a better dense gel structure and a higher stability, encapsulation rate, and antibacterial activity, suggesting that pH affects gel microstructure and thus the encapsulation efficiency and bacteriostatic properties of the prepared nanoparticles. These results contribute to our knowledge of the design and fabrication of polymeric nanoparticle delivery systems for bioactive compounds with beneficial properties.

Topics: Anti-Bacterial Agents; Bacillus cereus; Cymenes; Hydrogen-Ion Concentration; Microbial Sensitivity Tests; Nanocapsules; Nanogels; Ovalbumin; Particle Size; Rheology; Salmonella; Solubility

Th1, Th2, Th17, and Treg cells and their cytokine gene expressions in splenocytes of control mice, ovalbumin sensitized (S), and S treated with dexamethasone and carvacrol during a sensitization period were examined. Th2 and Th17 population as well as the gene expression of IL-4, IL-17, and TGF-β were increased, but Th1, Th1/Th2 ratio, the gene expression of IFN-γ and FOXP3 as well as the IFN-γ/IL-4 ratio were decreased in S compared with control group ( P < 0.001 for all cases). Carvacrol treatment caused significant reduction of Th2 and Th17 population as well as gene expression of IL-4, IL-17, and TGF-β but increase in Treg cells, Th1/Th2 ratio, gene expressions of FOXP3, IFN-γ, and IFN-γ/IL-4 ratio ( P < 0.05 to P < 0.001). The population of Th1, Th2, Th17 cells as well as the gene expression of IL-4, IL-17, and TGF-β were significantly decreased, but only Treg was increased in the dexamethasone treatment group ( P < 0.05 to P < 0.001). Carvacrol treatment during the sensitization period showed a more specific effect on Th1/Th2 imbalance in sensitized mice than dexamethasone, which may indicate the therapeutic potentials of carvacrol in disorders associated with Th1/Th2 imbalance such as asthma.

Topics: Animals; Asthma; Cymenes; Cytokines; Dexamethasone; Gene Expression; Gene Expression Regulation; Humans; Interferon-gamma; Interleukin-17; Interleukin-4; Mice; Ovalbumin; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells; Th2 Cells; Transforming Growth Factor beta

Thymol and carvacrol, two main components of thyme, have several valuable effects on the immune system. This study aims to evaluate the effects of these components on T-helper (TH) cell responses and their subsets in mice immunized with ovalbumin. The effects of these components on: a specific in vivo immune response were evaluated by assessing changes in delayed-type hypersensitivity (DTH); ex vivo splenocyte proliferative responses were evaluated using a BrdU assay gene expression of cytokines and key transcription factors involved in T-cells subset differentiation among the mouse splenocytes were assessed using real-time polymerase chain reaction (PCR); and splenocyte cytokine formation (ex vivo) and levels of the cytokines in mouse sera were measured by ELISA. Mice treated with thymol or carvacrol had reduced DTH responses (26% and 50%, respectively) compared with control mice. Thymol and carvacrol each diminished splenocyte proliferation to nearly 65-72% of control levels (p < 0.01). These agents also led to decreased TH1 [interleukin (IL)-2, interferon (IFN)-γ)], TH2 (IL-4) and TH17 (IL-17A) levels in the splenocyte cultures and in the sera of mice but increased levels of IL-10 and transforming growth factor (TGF)-β. Treated immunized mice showed significantly reduced T-box 21 (T-bet) expression from 3.8 [± 0.3]-fold in untreated ovalbumin-immunized mice to 0.9 [± 0.4]-(thymol) and 0.8 [± 0.2]-fold (carvacrol) (p < 0.01). GATA binding protein 3 (GATA-3) expression declined from 3.4 [± 0.4]- to 0.5 [± 0.3]-fold (thymol) and 0.6 [± 0.4]-fold (carvacrol), whereas RORγc decreased from 13.4 [± 1.6]- to 1.5 [± 0.6]-fold (thymol) and 0.8 [± 0.4]-fold (carvacrol) (p < 0.001). As carvacrol and thymol each suppressed the antigen-specific immune response by reducing TH cell-related cytokines\\specific transcription factors, this indicated their potential to modulate destructive immune responses attributed to T-cells over-activation.

Topics: Animals; Antigens; Cell Proliferation; Cells, Cultured; Cymenes; Cytokines; GATA3 Transcription Factor; Hypersensitivity, Delayed; Immunization; Male; Mice; Mice, Inbred BALB C; Monoterpenes; Nuclear Receptor Subfamily 1, Group F, Member 3; Ovalbumin; T-Box Domain Proteins; T-Lymphocyte Subsets; T-Lymphocytes, Helper-Inducer; Thymol; Thymus Plant; Transcriptional Activation

The effects of carvacrol on tracheal responsiveness (TR) to methacholine and ovalbumin (OA), serum nitric oxide (NO) concentration, total and differential white blood cells (WBC) in blood of sensitized guinea pigs were examined. Five groups of guinea pigs sensitized to OA were given drinking water alone (group S), drinking water containing three concentrations of carvacrol (40, 80 and 160 µg/mL) and dexamethasone (50 µg/mL). TR to methacholine and OA, serum NO concentration, total and differential WBC in blood of sensitized and control guinea pigs were measured (n = 6, for each group). TR to methacholine and OA, serum level of NO and nitrite, total WBC, eosinophil and neutrophil counts were increased but lymphocyte decreased in group S compared with control group (P < 0.01 for NO and nitrite and P < 0.001 for other cases). Treatment of S animals with dexamethasone and two higher concentrations of carvacrol significantly improved all measured parameters except TR to OA in treated group with dexamethasone (P < 0.05 to P < 0.001). Treatment of S animals with low concentration of carvacrol also improved TR to methacholine and OA, total WBC count and nitrite level (P < 0.05 to P < 0.001). The effects of two higher concentrations of carvacrol on TR, NO and nitrite and the effects of its highest concentration on total and differential WBC count were significantly higher than those of dexamethasone (P < 0.05 to P < 0.001). In addition, the effects of highest concentration of carvacrol on all parameters and its medium concentration on some parameters were significantly higher than its low concentration (P < 0.05 to P < 0.001). These results showed a preventive effect of carvacrol on tracheal responsiveness, serum level of NO and nitrite, total and differential WBC in the blood of sensitized guinea pigs which was equal or even more potent than dexamethasone at used concentrations.

Topics: Allergens; Animals; Cymenes; Dexamethasone; Eosinophils; Guinea Pigs; Immunization; Immunologic Factors; Leukocyte Count; Lymphocytes; Methacholine Chloride; Monoterpenes; Neutrophils; Nitric Oxide; Ovalbumin; Trachea