加味五虎追风散保护神经元的作用机制研究  

Study on mechanism for effect of protecting neuron with modified Wuhu zhuifeng powder

作　　者：梁健芬[1] 陈炜[1] 张兴博[1] 周华梅 黄少东 Liang Jian fen;Chen Wei;Zhang Xing bo;Zhou Hua mei;Huang Shao dong(The First Affiliated Hospitai of Guangxi University of Chinese Medicine,Nanning,Guangxi,53002)

机构地区：[1]广西中医药大学第一附属医院,广西南宁530023

出　　处：《广西中医药》2018年第5期55-58,共4页Guangxi Journal of Traditional Chinese Medicine

基　　金：广西自然科学基金项目(编号:2015GXNSFAA139226);广西中医基础研究重点实验室开放课题(编号:15-140-32-04)

摘　　要：目的:探讨加味五虎追风散对鱼藤酮诱导的帕金森病(PD)大鼠模型线粒体-ATP-mTOR信号通路和多巴胺(DA)能神经元自噬的影响。方法:将120只大鼠随机分为空白对照组、模型组、加味五虎追风散组和雷帕霉素靶蛋白(m TOR)抑制剂组4组,每组30只。空白对照组正常饲养,其他组均采用颈、背部交替皮下注射鱼藤酮法制备大鼠PD模型,加味五虎追风散组和m TOR抑制剂组分别以加味五虎追风散溶液和雷帕霉素溶液灌胃。干预3周后提取大鼠中脑和纹状体,采用线粒体呼吸测定仪测定线粒体呼吸控制比(RCR),采用荧光素-荧光素酶发光法测定线粒体ATP合成酶活力;采用免疫蛋白印迹实验测定磷酸化雷帕霉素靶蛋白(p-mTOR)、微管相关蛋白1轻链3B(LC3B)表达。结果:与空白对照组比较,模型组大鼠酪氨酸羟化酶(TH)基因表达、线粒体RCR、ATP合成酶活力及p-mTOR表达降低,LC3B蛋白表达升高(P<0.05或P<0.01);与模型组比较,加味五虎追风散组和m TOR抑制剂组TH表达、线粒体RCR、ATP合成酶活力及LC3B蛋白表达明显升高,p-m TOR表达明显降低(P<0.05或P<0.01)。结论:加味五虎追风散能够通过线粒体-ATP-mTOR途径上调DA能神经元自噬,发挥神经元保护作用。Objective： To investigate the effects of modified Wuhu zhuifeng powder on mitochondrial-ATP-mTOR signaling pathway and dopaminergic neuron autophagy in rats model of Parkinson＇s disease （PD） induced by rotenone. Methods： One-hundred and twenty rats were randomly divided into four groups： blank control group, model group, mod- ified Wuhu zhuife ng powder group and roTOR inhibitor group with 30 rats for each group. The blank control group was given normal feeding. In other groups, PD model rats were prepared by alternate subcutaneous injection of rotenone in the neck and back.Modified Wuhu zhuifeng powder group and roTOR inhibitor group were given by garage with modified Wuhu zhuife ng powder and rapamycin respectively. After 3 weeks of intmwention, the midbrain and striatum were extracted. And the mitochondrial respiration control ratio （RCR） was measured by the Mitochondrial Respiratory analyzer; the activity of mitochondrial ATP synthase was determined by fluorescence-luciferase assay; the expression of P-roTOR and LC3B was determined by immuno-protein imprinting assay. Results： The TH expression, mitochondrial RCR, ATP synthase activity and p-roTOR expression were decreased, and LC3B protein expression was increased in model group compared with those in blank control group. The TH expression, mitochondrial RCR, ATP synthase activity and LC3B protein expression were increased, and p-roTOR expression were decreased in modified Wuhu zhuifeng powder group and roTOR inhibitor group compared with those in model group. Conclusion： Modified Wuhu zhuifeng Powder can up-regulate DA neuronal autophagy and exert neuroprotective effect through the mitochondrial-ATP-mTOR pathway.

关 键 词：加味五虎追风散 帕金森 多巴胺 线粒体 自噬 

分 类 号：R285.5[医药卫生—中药学]