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Journal of ethnopharmacology

Bu-Shen-Yi-Qi formulae suppress chronic airway inflammation and regulate Th17/Treg imbalance in the murine ovalbumin asthma model.


PMID 25625352

Abstract

Bu-Shen-Yi-Qi formulae (BSYQF) are frequently used in the treatment of chronic inflammatory diseases in the respiratory system in traditional Chinese medicine (TCM). However, the regulatory effect of BSYQF on T helper 17 (Th17) and regulatory T (Treg) cells in murine ovalbumin (OVA) asthma model remains poorly understood. In the present study, we sought to determine the effect of high-performance liquid chromatography/mass spectrometry (HPLC/MS) standardized BSYQF on chronic airway inflammation and Th17/Treg imbalance in the murine OVA asthma model. The murine asthma model was induced by OVA sensitization and challenge and BSYQF was oral administrated. 24h after last OVA exposure, airway hyperresponsiveness (AHR) to methacholine (Mch) was assessed, and inflammatory cell counts and classification in bronchoalveolar lavage fluid (BALF) were analysed. Histopathological evaluation of the lung tissue was performed by hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) staining. Th17 and Treg associated cytokine levels in serum and BALF as well as transcription factors expression in the lung tissue were measured by ELISA, Bio-Plex and western blot assay. We also analysed the CD4(+)RORγt(+) and CD4(+)Foxp3(+) T cells in BALF and spleen by flow cytometric analysis. Our results demonstrated that oral administration of BSYQF inhibited the markedly increased AHR and lung inflammation (p<0.05), resulted in a dramatic reduction in total inflammatory cells as well as neutrophils (Neu), lymphocytes (Lym), monocytes (Mon), eosinophils (Eos) and basophils (Bas) of OVA-induced asthmatic mice (p<0.05). Furthermore, BSYQF treatment caused a distinct reduction in IL-6, IL-10 and IL-17A levels in serum (p<0.05), and induced a significant improvement in IL-6 and IL-10 as well as a marked decrease in TGF-β1 and IL-17A levels in BALF of OVA-induced asthmatic mice (p<0.05). Mice in BSYQF treated groups also had decreased RORγt and increased Foxp3 expression in the lung tissue (p<0.05). Flow cytometry analysis revealed that CD4(+)RORγt(+) T cells elevated markedly and CD4(+)Foxp3(+) T cells decreased prominently in BALF and spleen in murine OVA asthma model (p<0.05), and BSYQF and DEX treatment lead to an obvious reduction in CD4(+)RORγt(+) T cells in BALF (p<0.05) but not in spleen. BSYQF and DEX treatment resulted in an obvious elevation in CD4(+)Foxp3(+) T cells in BALF and spleen (p<0.05). Collectively, these results demonstrated that BSYQF could suppress chronic airway inflammation and regulate Th17/Treg imbalance by inhibition of Th17 and enhancement of Treg functions in the murine OVA asthma model, which may help to elucidate the underlying regulatory mode of BSYQF on asthma treatment.