加味五虎追风散通过调控TFEB介导的自噬干预帕金森病发病的实验研究  

Experimental Study of Modified Wuhu Zuifeng Powder Intervening the Pathogenesis of Parkinson′s Disease by Regulating TFEB-mediated Autophagy

作　　者：张兴博[1] 梁健芬[1] 陈炜[1] 徐柳炎 蒙健林 ZHANG Xingbo;LIANG Jianfen;CHEN Wei;XU Liuyan;MENG Jianlin(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,Guangxi,China)

机构地区：[1]广西中医药大学第一附属医院,南宁530023

出　　处：《中西医结合心脑血管病杂志》2021年第5期758-760,共3页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：广西自然科学基金面上项目(No.2018GXNSFAA138034)。

摘　　要：目的探讨转录因子EB(TFEB)介导的自噬在帕金森病(PD)发病中的作用和机制,从细胞自噬水平阐明加味五虎追风散的效用机制。方法将60只PD大鼠随机分为正常组、模型组、中药组,各20只。模型组及中药组大鼠每日按时颈部、背部交替皮下注射鱼藤酮葵花油乳化液建立PD大鼠模型。造模成功后中药组给予加味五虎追风散灌胃,正常组及模型组给予等体积蒸馏水灌胃。4周后处死大鼠,取脑部黑质组织,分离各组大鼠多巴胺能神经元,免疫印迹法(Western Blot)检测TFEB、α-突触核蛋白(α-syn)、微管相关膜蛋白轻链3Ⅱ(LC3Ⅱ)、泛素结合蛋白P62(P62)、酪氨酸羟化酶(TH)蛋白表达。结果与正常组比较,模型组大鼠α-syn、LC3Ⅱ、P62表达量增加,TH表达量减少,差异均有统计学意义(P<0.05或P<0.01),模型组与正常组比较,TFEB表达量无明显变化,差异无统计学意义(P>0.05);与模型组比较,中药组大鼠TFEB、LC3Ⅱ、TH表达量明显增加,P62、α-syn表达量明显减少,差异均有统计学意义(P<0.05或P<0.01)。结论加味五虎追风散可通过调控TFEB介导的自噬减少α-syn异常聚集,增加TH表达,从而发挥神经保护作用。Objective To clarify the role and mechanism of transcription factor EB(TFEB)-mediated autophagy in the pathogenesis of Parkinson′s Disease(PD),and to elucidate the action mechanism of Jiawei Wuhu Zhuifeng powder from the level of cellular autophagy.Methods Sixty PD rats were randomly divided into three groups:normal group,model group,and Chinese medicine group,with 20 rats in each group.Rats in model group and Chinese medicine group were injected subcutaneously with rotenone and sunflower oil emulsions to establish a PD rat model.After the successful modeling,the rats in Chinese medicine group were given gavage of Wuhu Zhuifeng powder,and the rats in normal group and model group were given the same volume of distilled water.After 4 weeks,the rats were sacrificed,and the substantia nigra tissues were collected.The dopamine(DA)neurons in each group were isolated.The TFEB,α-synuclein(α-syn),microtubule-associatedn protein-light chain 3Ⅱ(LC3Ⅱ),P62,and tyrosine hydroxylase(TH)protein expression were detected.Results Compared with normal group,the expression ofα-syn,LC3Ⅱ,and P62 in model group increased,and the expression of TH decreased(P<0.05 or P<0.01).The difference in the expression of TFEB was not statistically significant between model group and normal group(P>0.05).Compared with model group,the expression levels of TFEB,LC3Ⅱ,and TH in Chinese medicine group increased significantly,and the expression levels of P62 andα-syn in Chinese medicine group decreased significantly,the difference was statistically significant(P<0.05 or P<0.01).Conclusion Jiawei Wuhu Zhuifeng powder can reduce TFEB mediated autophagy to reduceα-syn abnormal aggregation and increase TH expression to exert neuroprotective effect.

关 键 词：帕金森病 转录因子EB 多巴胺能神经元 自噬 加味五虎追风散 Α-突触核蛋白 微管相关膜蛋白轻链3Ⅱ 酪氨酸羟化酶 

分 类 号：R73[医药卫生—肿瘤]