线粒体分裂蛋白抑制剂对匹鲁卡品致痫大鼠海马神经元的保护作用

Effect of mitochondrial division inhibitor on hippocampal neuro in pilocarpine-induced seizures of rats

目的:研究线粒体分裂蛋白抑制剂(Mdivi-1)对匹鲁卡品(PILO)致痫大鼠海马神经元的保护作用及其机制。方法:121只雄性SD大鼠随机分成对照(CON)组、PILO组、PILO+DMSO组和PILO+Mdivi-1组,PILO组又分为癫痫持续状态(SE)2 h、8 h、24 h和72 h 4个亚组。观察各组大鼠行为学改变,并分别采用Nissl染色法检测神经元损伤,比色法检测SOD活力及MDA含量,Western blot和RT-PCR法检测Drp1、细胞色素C(Cyt C)、凋亡诱导因子(AIF)和cleaved-半胱天冬酶3(caspase-3)表达水平。结果:与PILO组(114.571±5.420)相比,PILO+Mdivi-1组海马神经元数目增加[(157.429±6.183);F=68.693,P<0.001]。与PILO组SE 24 h亚组[(113.429±4.980),(6.102±0.193)]相比,PILO+Mdivi-1组SOD活力增加[(138.571±3.380);F=21.779,P<0.001],MDA含量减少[(4.353±0.231);F=27.226,P<0.001]。与PILO组[(0.594±0.020),(1.099±0.015)]相比,PILO+Mdivi-1组线粒体Cyt C增加[(0.897±0.015);F=296.177,P<0.001],细胞质Cyt C减少[(0.433±0.010);F=562.684,P<0.001]。与PILO组[(0.878±0.037),(1.465±0.076)]相比,PILO+Mdivi-1组线粒体AIF增加[(1.279±0.051);F=59.298,P<0.001],细胞核AIF减少[(0.896±0.044);F=46.424,P<0.001]。与PILO组(0.587±0.192)相比,PILO+Mdivi-1组cleaved caspase-3减少[(0.463±0.022);F=40.500,P<0.001]。结论:Mdivi-1对癫痫大鼠海马神经元有保护作用,其机制可能与降低氧化应激,抑制Cyt C释放、AIF移位及caspase-3激活有关。

Aim：To investigate the effect of mitochondrial division inhibitor （ Mdivi1-） in pilocarpine-induced seizures and its mechanism .Methods：A total of 121 male SD rats were randomly divided into control , PILO,PILO＋Mdivi-1 and PILO＋DMSO group, the PILO group was further divided into four subgroups according to observation time .The behavior changes of each group were observed , while the pathological damage and levels of superoxide dismutase and malondialde-hyde were detected by Nissl staining ,xanthine oxidase and thiobarbituric acid method .The expression of dynaminr-elated protein 1,Cyt C , AIF and cleaved-caspase-3 were measured by Western blot analysis and RT-PCR.Results： Compared with PILO group （114.571 ±5.420）, the surviving neuron number of PILO ＋Mdivi-1 group was increased [（157.429 ± 6.183）;F=68.693,P〈0.001].Compared with PILO SE 24 h subgroup [（113.429 ±4.980）,（6.102 ±0.193）],the activity of SOD[（138.571 ±3.380）;F=21 .779,P〈0.001]was increased and the content of MDA （4.353 ±0.231）;F=27.226,P〈0.001]was decreased in PILO＋Mdivi-1 group.Compared with PILO group[（0.594 ±0.020）,（1.099 ± 0.015）], the expression of mito-Cyt C[（0.897 ±0.015）;F=296.177,P〈0.001]was increased and the expression of cyto-Cyt C[（0.433 ±0.010）;F=562.684,P〈0.001] was decreased in PILO ＋Mdivi-1 group.Compared with PILO group[（0.878 ±0.037）,（1.465 ±0.076）], the expression of mito-AIF[（1.279 ±0.051）;F=59.298,P〈0.001]was increased and the expression of nu-AIF[（0.896 ±0.044）;F =46.424,P 〈0.001] was decreased in PILO ＋Mdivi-1 group.Compared with PILO group （0.587 ±0.192）, the expression of cleaved caspase-3[（0.463 ±0.022）;F=40.500, P〈0.001]was decreased of PILO ＋Mdivi-1 group.Conclusion：Mdivi-1 has neuroprotective effect on hippocampal neuro in pilocarpine-induced seizures , and the underlying mechanism may be due to the suppression of oxidative stress and inhibi -tion of the release of Cyt C , translocation of AIF and activation of caspase-3.