祛风宣肺汤治疗冷刺激联合屋尘螨诱导哮喘小鼠的作用机制

Mechanism of the Therapeutic Effect of Qufeng Xuanfei Decoction on Cold-stimulating and House Dust Mite Induced Asthmatic Mice

目的:探讨祛风宣肺汤治疗冷刺激联合屋尘螨(house dust mite,HDM)诱导的过敏性哮喘小鼠的作用机制。方法:40只雄性BALB/c小鼠随机分为5组,即空白组、冷刺激组、HDM组、模型(冷刺激+HDM)组和祛风宣肺汤(11.83 g·kg^(-1))组,每组8只。采用反复低温(10±3℃)刺激加HDM建立小鼠过敏性哮喘模型。给药24 d后,检测支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中白细胞介素-5(interleukin-5,IL-5)、IL-13的水平、外周血嗜酸性粒细胞(eosinophil,EOS)计数及血清免疫球蛋白E(immunoglobulin E,IgE)水平;HE染色观察肺组织病理;免疫组化法检测瞬时肺组织受体电位M8亚型通道蛋白(transient receptor potential melastatin 8,TRPM8)的表达。结果:与空白组相比,冷刺激组、HDM组、模型组BALF中IL-5和IL-13水平、外周血EOS计数和血清IgE水平均升高;病理结果显示肺泡间隙及毛细血管周围大量炎性细胞浸润;免疫组化结果显示肺组织中TRPM8表达增加,以模型组效果最为明显。与模型组相比,祛风宣肺汤组BALF中IL-5、IL-13水平、外周血EOS计数、血清IgE水平均明显降低,且肺组织炎症浸润及病变程度明显减轻,TRPM8蛋白表达明显降低。结论:祛风宣肺汤能降低小鼠肺组织TRPM8蛋白表达水平,调节Th2型免疫应答,改善肺组织炎症浸润,达到缓解过敏性哮喘的作用。

Objective:To explore the mechanism of Qufeng Xuanfei Decoction on asthmatic mice induced by cold air and house dust mite(HDM).Methods:40 male BALB/c mice were randomly divided into 5 groups:the control group,cold stimulation group,HDM group,model(cold stimulation+HDM)group,and Qufeng Xuanfei Decoction(11.83 g·kg^(-1))group,with 8 mice in each group.A mouse model of allergic asthma was established by repeated low temperature(10±3℃)stimulation and HDM nasal drops.After 24 days of the administration,detect the levels of interleukin-5(IL-5)and IL-13 in bronchoalveolar lavage fluid(BALF)and eosinophil(EOS)counts in peripheral blood and serum immunoglobulin E(IgE)levels.HE staining was used to observe lung tissue pathology,and immunohistochemistry was used to observe the expression of transient receptor potential melastatin 8(TPRPM8)in lung tissue.Results:Compared with the control group,the levels of IL-5 and IL-13 in the BALF,the peripheral blood EOS count,and the serum IgE titer in the cold stimulation group,HDM group,and model group was increased.The pathological results showed a large number of inflammatory cell infiltration in the alveolar space and around the capillaries.Immunohistochemical results showed increased expression of TRPM8 in lung tissue,and the model group has the best replication.Compared with the model group,the levels of IL-5,IL-13,EOS count in peripheral blood,and serum IgE titer in the BALF of the Qufeng Xuanfei Decoction group were significantly reduced,and the degree of inflammation and lesions in the lung tissue was significantly reduced,and the expression of TRPM8 protein reduced.Conclusion:Qufeng Xuanfei Decoction can reduce the expression of TRPM8 protein in lung tissue of mice,regulate Th2-type immune response,improve lung tissue inflammation and infiltration,and achieve the effect of alleviating allergic asthma.