基于“肺阳虚”理论探析温阳化饮方对哮喘寒饮蕴肺证细胞自噬的作用机制

Study on the mechanism of Wenyang Huayin Decoction on autophagy of asthma cells with cold yin accumulating lung syndrome based on the theory of‘lung yang deficiency’

目的:基于“肺阳虚”理论通过温阳化饮方干预哮喘寒饮蕴肺证大鼠模型,研究温阳化饮方对哮喘寒饮蕴肺证细胞自噬的作用机制。方法:72只Wistar大鼠随机分为空白组、模型组、小青龙汤组、温阳化饮方组、黄芪多糖组、人参皂苷组,每组12只。通过构建哮喘寒饮蕴肺证大鼠模型,并分别给予小青龙汤、温阳化饮方、黄芪多糖、人参皂苷进行治疗,引入HE染色法观察肺组织病理改变;透射电镜观察肺组织自噬小体情况;酶联免疫吸附剂测定IL-18、IFN-γ等相关炎症因子表达水平;逆转录-聚合酶链反应检测肺组织自噬相关基因ATG mRNA的表达水平。结果:与空白组比较,模型组支气管黏膜上皮细胞出现严重增生、肿胀、脱落、坏死等病理表现,并伴有较多炎性细胞浸润;炎症因子IL-18、IFN-γ的表达水平有着明显升高的趋势(P<0.01),自噬相关基因ATG3、ATG7 mRNA的表达水平显著降低(P<0.01,P<0.05),各治疗组较模型组上述指标显著改善,其中温阳化饮方组效果最佳。结论:“肺阳虚”会影响细胞自噬的总体水平,导致哮喘寒饮蕴肺证的发病或病情加重,温阳化饮方可调节细胞自噬水平,使其趋于正常,从而降低哮喘炎症水平,这可能是温阳化饮方治疗哮喘寒饮蕴肺证的作用机制。

Objective:To study the mechanism of wenyang Huayin Decoction(WYHY)on the syndrome of cold yin accumulating lung in asthma,and to explore the overall effect of the state of‘lung yang deficiency’on the level of autophagy.Methods:A total of 72 Wistar rats were randomly divided into the blank group,the model group,the Xiaoqinglong Decoction group,the WYHY group,the astragalus polysaccharide group and the ginsenoside group,12 rats in each group.The rat model of asthma cold yin accumulating lung syndrome was established and treated with Xiaoqinglong Decoction,WYHY,astragalus polysaccharide and ginsenoside respectively.HE staining was used to observe the pathological changes of lung tissue;transmission electron microscopy was used to observe the autophagy of lung tissue;enzyme-linked immunosorbent assay was used to detect the expression level of IL-18,IFN-γand other related inflammatory factors;reverse transcription polymerase chain reaction was used The expression of ATG mRNA was detected.Results:Compared with the blank group,the model group showed inflammatory cell infiltration;the expression level of inflammatory factors IL-18,IFN-γwere significantly increased(P<0.01);the expression level of ATG3,ATG7 mRNA related to autophagy was significantly reduced(P<0.01,P<0.05);the above indicators of each treatment group were significantly improved compared with the asthma model group,and the effect of WYHY group was the best(P<0.01,P<0.05).Conclusion:‘Deficiency of lung yang’can affect the overall level of autophagy,and lead to the pathogenesis or aggravation of asthma cold yin lung syndrome.WYHY can regulate the autophagy level of cells,so that it tends to be normal,so as to reduce the level of asthma inflammation,which may be the mechanism of WYHY in the treatment of asthma cold yin accumulating lung syndrome.