Abstract
Purpose: To investigate the anti-asthmatic effects of the flavonoid rutin in an ovalbumin (OVA)-induced asthmatic mouse model. Methods: Asthma was induced by OVA induction. Effects of rutin (25–100 mg/kg BW) on interleukins (IL)-4, IL-5, and IL-13 and interferon (IFN)-γ in bronchoalveolar lavage fluid (BALF) and serum OVAspecific IgE levels were assessed by enzyme linked immunosorbent assay (ELISA). Airway hyperresponsiveness (AHR) was measured by determining lung compliance and airway resistance. Expression levels of p-I-κBα, p-NF-κB p65, TNF-α, and Foxp3 were also assessed. The CD4 + CD25 + Foxp3 + T regulatory cell population was determined by flow cytometry. Results: Rutin significantly ( p < 0.05) decreased Th2 cytokines (IL-4, IL-5, IL-13), whereas it enhanced IFN-γ levels in BALF. Decreased OVA-specific serum IgE was also observed with improved lung histology. Rutin was effective in reducing AHR and regulating the expression levels of p-I-κBα, p-NF-κB p65, and TNF α, and also increased the CD4 + CD25 + Foxp3 + Treg cell population and Foxp3 mRNA and protein levels. Conclusion: Rutin effectively suppresses OVA-induced asthma and improves airway function by suppressing inflammatory mediators and inflammatory cell infiltration. Keywords: Asthma, CD4 + CD25 + , Cytokines, Inflammation, Ovalbumin, Rutin, Protein levels
Highlights
Allergic asthma is a chronic airway disorder characterised by airway inflammation, airway hyperresponsiveness (AHR), and mucus hypersecretion [1]
OVA-sensitised mice that were treated with rutin at 100 mg showed reduced eosinophil and neutrophil counts versus mice treated with lower doses of rutin
We examined the population of CD4+ CD25+ Foxp3+ Tregs in bronchoalveolar lavage fluid (BALF) of OVA-challenged mice
Summary
Allergic asthma is a chronic airway disorder characterised by airway inflammation, airway hyperresponsiveness (AHR), and mucus hypersecretion [1]. On the days of OVA challenge, the mice received rutin 60 min before injections. A separate group of mice received dexamethasone (Dex; 2 mg/kg) i.p. 1 h before OVA injections. OVA-specific serum IgE levels were determined using the Legend Max mouse OVA-specific IgE ELISA kit (BioLegend, San Diego, CA, USA) according to the manufacturer’s protocol. The inflammatory scores and expression levels of mucus-positive cells were calculated as described previously [17]. For Treg analysis, cells were incubated with FITC-labelled anti-CD4 and APC-labelled anti-CD25 antibodies in staining buffer and incubated for 30 min at 4 °C. The cells were fixed, permeabilised, and subsequently stained with anti-rat Foxp and resuspended in staining buffer and subjected to analysis by flow cytometry (FACSCalibur instrument with CellQuest software; BD Biosciences, Mountain View, CA, USA). Data were analysed for statistical significance at p < 0.05 by one-way analysis of variance (ANOVA) followed by Duncan’s multiple range test (DMRT) as a post hoc analysis
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