过表达E46K突变体α-突触核蛋白抑制线粒体自噬的研究  

Overexpression of E46K Mutant α-Synuclein Inhibits Mitophagy

作　　者：闫加庆 苑玉和[2] 楚世峰[2] 刘敏 马颖林 乐凯迪 李国辉 陈乃宏[2] YAN Jia-qing;YUAN Yu-he;CHU Shi-feng;LIU Min;MA Ying-lin;LE Kai-di;LI Guo-hui;CHEN Nai-hong(Department of Pharmacy,National Cancer Center,National Clinical Research Center for Cancer,Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,China;Neuroscience Center,Institute of Materia Medica,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100050,China)

机构地区：[1]国家癌症中心,国家肿瘤临床医学研究中心,中国医学科学院北京协和医学院肿瘤医院药剂科,北京100021 [2]中国医学科学院北京协和医学院药物研究所神经科学中心,北京100050

出　　处：《中国药学杂志》2021年第15期1228-1231,共4页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金项目资助(82003973);北京市优秀人才培养资助青年骨干个人项目资助(2018000032600G402)。

摘　　要：目的研究E46K突变体α-突触核蛋白(α-synuclein,α-syn)对线粒体自噬的影响。方法构建稳定表达E46K突变体α-syn的PC12细胞株,检测细胞内活性氧含量和细胞凋亡的发生,并通过western blot和免疫共沉淀方法检测线粒体自噬相关蛋白Parkin、PINK1的细胞内定位和相互作用情况。结果本研究成功构建稳定表达E46K突变体α-syn的细胞株。在线粒体解偶联剂CCCP作用下,过表达E46K突变体α-syn可导致细胞内活性氧含量和细胞凋亡显著增加,并抑制线粒体自噬蛋白Parkin由细胞浆向线粒体募集,导致Parkin与PINK1相互作用减少。结论过表达E46K突变体α-syn可能通过抑制线粒体自噬发生促进帕金森病(PD)的发生和发展。OBJECTIVE To explore the effects of E46 K mutant α-synuclein(α-syn) on mitophagy. METHODS Stable PC12 cells lines expressing E46 K mutant α-syn were constructed. The levels of reactive oxygen species(ROS) and apoptosis rate were detected by flow cytometry, and the levels and location of mitophagy-related proteins such as PINK1 and Parkin were evaluated by Western blot and co-immunoprecipitation assays. RESULTS Overexpression of E46 K mutant α-syn could significantly increase ROS levels and apoptosis rate under CCCP treatment. Otherwise, expressing E46 K mutant α-syn inhibited translocation of Parkin from cytoplasm to mitochondrial and interaction with PINK1. CONCLUSIONS Overexpression of E46 K mutant α-syn may promote PD development by inhibiting mitophagy.

关 键 词：Α-突触核蛋白 E46K突变体 线粒体自噬 

分 类 号：R966[医药卫生—药理学]