线粒体自噬抑制剂Mdivi-1调控线粒体未折叠蛋白反应改善癫痫海马神经元损伤  

The mitochondrial autophagy inhibitor Mdivi-1 improve the injury of epileptic hippocampal neurons by regulating mitochondrial unfolded protein response

作　　者：谢南昌[1] 杜丽媛 连亚军[1] 王翠[2] 孟祥荷 李钰娟 刘凤霞 XIE Nanchang;DU Liyuan;LIAN Yajun(Department of Neurology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院神经内科,河南郑州450052 [2]郑州大学第一附属医院检验科,河南郑州450052

出　　处：《中风与神经疾病杂志》2020年第12期1087-1090,共4页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金资助项目(No.81971214,No.81701272);河南省医学科技攻关计划省部共建青年项目(No.SB201902011);郑州大学青年骨干教师培养计划培养人选(No.2019ZDGGJS056)。

摘　　要：目的探讨线粒体未折叠蛋白反应在无镁外液致痫海马神经元中的变化及线粒体自噬抑制剂Mdivi-1对其影响。方法原代培养海马神经元用无镁外液诱导体外癫痫模型,并进一步采用线粒体自噬抑制剂Mdivi-1干预;采用线粒体荧光探针MitoSOX检测线粒体ROS生成、Rhodamine 123检测线粒体膜电位、采用Western blotting方法观察C/-EBP同源蛋白CHOP、线粒体内热休克蛋白HSP60、线粒体蛋白酶ClpP表达、CytC释放和caspase-3激活。结果(1)与对照组相比,致痫组海马神经元CHOP、HSP60和ClpP表达明显增加;与致痫组相比,线粒体自噬抑制剂Mdivi-1可明显增加海马神经元CHOP、HSP60和ClpP表达;(2)与对照组相比,致痫组海马神经元线粒体ROS生成增多,并且线粒体膜电位降低;与致痫组相比,线粒体自噬抑制剂Mdivi-1可明显减少线粒体ROS生成,增加线粒体膜电位;(3)与对照组相比,致痫组海马神经元CytC释放及caspase-3激活增加;与致痫组相比,线粒体自噬抑制剂Mdivi-1可明显减少CytC释放及caspase-3激活。结论癫痫海马神经元线粒体未折叠蛋白反应激活,而抑制线粒体自噬可通过增强线粒体未折叠蛋白反应而发挥癫痫保护作用。Objective To investigate the changes of mitochondrial unfolded protein response in epileptic hippocampal neurons induced by Mg2+-free solution and the effects of mitochondrial autophagy inhibitors Mdivi-1.Methods Epilepsy model was induced in primary cultured hippocampal neurons of SD rats in vitro by Mg2+-free solution,and further treated with mitochondrial autophagy inhibitor Mdivi-1.Mitochondrial ROS production were detected by MitoSox.The mitochondrial membrane potential was detected by Rhodamine123.The expression of C/EBP homologous protein CHOP,mitochondrial heat shock protein HSP60,mitochondrial protease ClpP,CytC release and caspase-3 activation were detected by Western blotting.Results(1)Compared with the control group,the expression of CHOP,HSP60 and ClpP in the AE group was significantly increased.Compared with the AE group,Mdivi-1 pretreatment enhanced the expression of CHOP,HSP60 and ClpP.(2)Compared with the control group,the production of ROS in the AE group increased,and the mitochondrial menmbrane potential decreased.Compared with the AE group,Mdivi-1 pretreatment significantly decreased the production of mitochondrial ROS and increased the mitochondrial membrane potential.(3)Compared with the control group,the release of CytC and the activation of caspase-3 in the AE group increased.Compared with the AE group,Mdivi-1 pretreatment decreased the release of CytC and caspase-3 activation.Conclusion Mitochondrial unfolded protein response is activated in epileptic hippocampal neurons,while inhibition of mitochondrial autophagy may play a protective role via enhancing mitochondrial unfolded protein response.

关 键 词：线粒体未折叠蛋白反应 线粒体自噬 癫痫 Mdivi-1 

分 类 号：R741.1[医药卫生—神经病学与精神病学]