补肾益肺消癥方抑制肺成纤维细胞线粒体自噬与调控JNK/p62/Parkin通路机制的研究  

Study on mechanism of inhibiting mitochondrial autophagy and regulating JNK/p62/Parkin pathway of lung fibroblast by Bushen Yifei Xiaozheng Fang

作　　者：马乾[1] 马雪颜 何毓玺 马林沁 李艳鹏[1] 毕玮 张佳琪 刘艺 吴彩军 高伟华 张沂 果海凤 晏军[1] MA Qian;MA Xue-yan;HE Yu-xi;MA Lin-qin;LI Yan-peng;BI Wei;ZHANG Jia-qi;LIU Yi;WU Cai-jun;GAO Wei-hua;ZHANG Yi;GUO Hai-feng;YAN Jun(Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China;Beijing University of Chinese Medicine,Beijing 100029,China;Chongqing Hospital of Chinese Medicine,Chongqing 400021,China;Beijing University of Chinese Medicine Third Affiliated Hospital,Beijing 100029,China)

机构地区：[1]北京中医药大学东直门医院(第一临床医学院),北京100700 [2]北京中医药大学,北京100029 [3]重庆市中医院,重庆400021 [4]北京中医院大学第三附属医院,北京100029

出　　处：《中华中医药杂志》2022年第4期2203-2206,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：北京中医药大学2019年度校级课题青年教师项目(No.2019-JYB-JS-062)。

摘　　要：目的:探究补肾益肺消癥方干预特发性肺间质纤维化(IPF)进展的可能机制,为“肺络微型癥瘕”理论在中医药防治IPF奠定客观基础。方法:通过TGF-β1诱导MRC-5细胞转分化肺成纤维细胞,制备补肾益肺消癥方冻干粉,激光共聚焦显微镜观察MRC-5细胞线粒体自噬体数量及状况,通过Western Blot法检测NOX4、JNK、p62、Parkin、PINK1蛋白的表达。结果:模型组在TGF-β1刺激后较正常细胞组线粒体自噬现象明显增多,而西药对照组及中药低、中、高浓度组线粒体自噬现象较模型组减少。与正常组比较,模型组TGF-β1可显著激活JNK/p62/Parkin信号通路,各给药组NOX4、JNK、p62表达显著降低(P<0.05),Parkin、PINK1表达显著升高(P<0.05),但中药中浓度组对于降低JNK的表达明显优于其他给药组(P<0.05)。结论:补肾益肺消癥方治疗IPF的机制之一可能是通过抑制细胞线粒体自噬功能与调控JNK/p62/Parkin通路减缓肺组织的纤维化进程。Objective:To investigate the possible mechanism of Bushen Yifei Xiaozheng Fang in the treatment of idiopathic pulmonary fibrosis(IPF),laying an objective foundation for the theory of‘micro-zhengjia of lung collateral’in the prevention and treatment of IPF by TCM.Methods:MRC-5 cells were induced to transdifferentiate into pulmonary muscle fibroblasts by TGF-β1,and the lyophilized powder of Bushen Yifei Xiaozheng Fang was prepared.The number and status of mitochondrial autophagosome in MRC-5 cells were observed by laser confocal microscopy,and the protein expressions of NOX4,JNK,P62,Parkin and PINK1 were detected by Western Blot.Results:Model group cells can be observed after TGF-β1 stimulate mitochondrial autophagy significantly increased,while the western medicine group,TCM low,medium and high dose group after the intervention mitochondrial autophagy decreased significantly.Compared with the normal control group,the model group could significantly activate the JNK/p62/Parkin signaling pathway,and all doses TCM groups and Western medicine control group could inhibit the activation of JNK/p62/Parkin signaling pathway by decreasing the expressions of NOX4,JNK and p62(P<0.05),increasing the expressions of Parkin and PINK1(P<0.05).However,the JNK expression was significantly decreased in the TCM medium dose group than in other dose medicine groups(P<0.05).Conclusion:One of the mechanisms of Bushen Yifei Xiaozheng Fang in the treatment of idiopathic pulmonary fibrosis,may be through inhibiting the autophagy function of mitochondria,and regulating JNK/p62/Parkin signaling pathways to slow down the fibrosis process of lung tissues.

关 键 词：补肾益肺消癥方 MRC-5细胞 线粒体自噬 JNK/p62/Parkin通路 

分 类 号：R285[医药卫生—中药学]