OGD动态处理后大鼠海马神经元线粒体裂解及凋亡相关蛋白的变化  

Oxygen and glucose deprivation alters mitochondria dynamic and apoptosis-related proteins in a time-dependent manner

作　　者：李学涵 刘学良 王平 吕志宇 吴勇 陈秀 LI Xue-han;LIU Xue-liang;WANG Ping;LüZhi-yu;WU Yong;CHEN Xiu(Neurology Department,Southwest Medical University Affiliated Hospital,Luzhou 646000;Department of Geriatrics,Traditional Medical Hospital of Xindu,Chengdu 610500,China)

机构地区：[1]西南医科大学附属医院神经内科,四川泸州646000 [2]成都市新都区新都中医医院老年病科,四川成都610500

出　　处：《西安交通大学学报（医学版）》2018年第6期793-797,820,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：四川省教育厅自然科学研究资助项目(No.08zc036)~~

摘　　要：目的大鼠海马神经元经过不同时间糖氧剥夺(oxygen and glucose deprivation,OGD)处理后,观察其线粒体形态改变与凋亡相关蛋白的变化。方法将原代培养大鼠海马神经元随机分为5组:空白对照组(对照组)与OGD处理30min组、45min组、60min组及90min组,采用激光共聚焦显微镜观察线粒体形态;JC-1检测线粒体膜电位变化;Western blot检测胞质内磷酸化线粒体动力相关蛋白(P-Drp1蛋白)、细胞色素C(CytC)及含半胱氨酸的天冬氨酸水解蛋白3(caspase 3)蛋白的表达。结果随着OGD处理时间延长,(1)P-Drp1蛋白表达下降,线粒体裂解率逐渐增加(P<0.05);(2)线粒体膜电位下降越严重;(3)与对照组相比,60min组及90min组CytC、caspase 3蛋白的表达显著增加(P<0.05)。结论随着OGD处理延长,线粒体裂解增加,膜电位下降,膜通透性增加,且在OGD处理60min时伴随凋亡发生。Objective To evaluate the progressive effect of oxygen and glucose deprivation on fission level of mitochondria,transformation of fission-related protein and ultimately apoptosis-related biogenesis in rat hippocampal neurons.Methods Primary cultured neurons were divided into 5 groups randomly:normal control group(OGD 0min,0min),OGD 30min group(30min),OGD 45min group(45min),OGD 60min group(60min),and OGD 90min group(90min).The neurons were exposed to oxygen and glucose deprivation(OGD)for 0min,30min,45min,60min and 90min.Mitochondria were stained by Mitochondria tracker Deep Red and shot by laser confocal microscopy,and mitochondrial membrane potential was stained by JC-1 and examined by fluorescence microscopy.Assays of P-Drp1,CytC and caspase 3 were quantified by Western blot.Results①With prolonged OGD treatment,P-Drp1 protein expression in neurons was obviously decreased and mitochondrial fission rate gradually increased(P<0.05).②Mitochondrial transmembrane potential decreased more greatly.③Cytochrome C(CytC)and caspase 3 proteins had significantly high expression in 60min and 90min groups as compared with control group(P<0.05).Conclusion As OGD treatment prolongs,mitochondrial fission increases,mitochondrial transmembrane potential decreases,and membrane permeability increases,which occur with apoptosis at OGD 60 minutes time point of OGD treatment.

关 键 词：大鼠海马神经元 糖氧剥夺处理 Drp1蛋白 CYTC caspase 3 

分 类 号：R743.3[医药卫生—神经病学与精神病学]