线粒体未折叠蛋白反应在癫痫海马神经中的变化及线粒体特异性抗氧化剂对其影响  

The changes of mitochondrial unfolded protein response in epileptic hippocampal neurons and the effect of mitochondria-targeted antioxidant Mito-TEMPO

作　　者：谢南昌[1] 于晓梦 王晓艺 杜丽媛 刘凤霞 张婉婉 连亚军[1] XIE Nanchang;YU Xiaomeng;WANG Xiaoyi(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》2022年第5期414-418,共5页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金资助项目(81971214)。

摘　　要：目的观察线粒体未折叠蛋白反应(mitochondrial unfolded protein response,mtUPR)在氯化锂-匹鲁卡品(pilocarpine,PILO)致痫大鼠海马神经中的变化及线粒体特异性抗氧化剂Mito-TEMPO对其影响。方法采用PILO诱导癫痫大鼠模型,并进一步采用线粒体特异性抗氧化剂Mito-TEMPO进行干预;将成年雄性Wistar大鼠随机分为空白对照组(CON组)、致痫组(PILO组)、Mito-TEMPO组和PILO+Mito-TEMPO组;采用Nissl染色观察海马神经元损伤,电子透射显微镜观察线粒体超微结构,活性氧荧光探针(DCFDA)检测线粒体ROS生成,Rhodamine123染色法检测线粒体膜电位变化,Western blot法检测线粒体热休克蛋白HSP60、蛋白酶LONP1、线粒体蛋白酶CLpP的表达。结果(1)与CON组相比,PILO组海马神经线粒体超微结构破坏严重,线粒体ROS生成增多,线粒体膜电位降低;(2)与CON组相比,PILO组海马神经HSP60、LONP1和CLpP表达增加;(3)与PILO组相比,PILO+Mito-TEMPO组线粒体超微结构破坏减轻,线粒体ROS生成明显减少,线粒体膜电位增高;(4)与PILO组相比,PILO+Mito-TEMPO组海马神经HSP60、LONP1和CLpP表达降低。结论mtUPR在癫痫海马神经损伤中明显激活,Mito-TEMPO可能通过调控mtUPR对癫痫海马线粒体损伤发挥保护作用。Objective To investigate the changes of mitochondrial unfolded protein response(mtUPR)in hippocampus of lithium-pilocarpine induced seizures and the effect of mitochondria-targeted antioxidant.Methods Lithium chloride and pilocarpine were used to elicit status epilepticus in rats,and the rats were further treated with mitochondria-targeted antioxidant Mito-TEMPO.Adult male Wistar rats were randomly divided into control group,PILO group,Mito-TEMPO group and PILO+Mito-TEMPO group.Nissl staining was used to observe the damage of hippocampal neurons,electron transmission microscopy was used to observe the ultrastructure of mitochondria,ROS production of mitochondria was detected by reactive oxygen species fluorescent probe(DCFDA),and mitochondrial membrane potential was detected by Rhodamine123 staining.The expressions of mitochondrial heat shock protein HSP60,protease LONP1 and mitochondrial protease CLpP were detected by Western blot.Results(1)Compared with CON group,the ultrastructure of hippocampal neuronal mitochondria in PILO group was seriously damaged,mitochondrial ROS production was increased,and mitochondrial membrane potential was decreased.(2)Compared with the CON group,the expressions of HSP60,Lonp1 and CLpP in the PILO group were increased.(3)Compared with the PILO group,the damage of mitochondrial ultrastructure was alleviated in the PILO+Mito-TEMPO group,the production of mitochondrial ROS was significantly reduced,and the mitochondrial membrane potential was increased.(4)Compared with the PILO group,the expressions of HSP60,LONP1 and CLpP in the hippocampal neurons of the PILO+Mito-TEMPO group were decreased.Conclusion The mtUPR is significantly activated in hippocampal neurons in epilepsy,and Mito-TEMPO may play a protective role in hippocampal mitochondrial injury in epilepsy by regulating mtUPR.

关 键 词：癫痫 氧化应激 线粒体特异性抗氧化剂 线粒体未折叠蛋白反应 

分 类 号：R742.1[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学]