线粒体分裂抑制剂通过抑制自噬加重OGD/R损伤  

Mitochondrial fission inhibitor aggravates OGD/R-induced injury through inhibition of autophagy

作　　者：彭程[1] 张磊[1] 饶维[1] 费舟[1] 

机构地区：[1]第四军医大学西京医院神经外科,陕西西安710032

出　　处：《中华神经外科疾病研究杂志》2016年第2期101-104,共4页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(81430043)

摘　　要：目的探讨线粒体分裂抑制剂(Mdivi-1)处理对原代培养的皮层神经元缺血再灌注损伤的作用及其机制研究。方法在原代培养的小鼠皮层神经元上建立氧糖剥夺/复氧复糖(OGD/R)模型模拟体内缺血再灌注损伤,根据再灌注时间长短检测自噬的发生情况,并进一步选取自噬发生最明显的时间点。然后分别采用自噬抑制剂-3-MA、自噬激动剂-雷帕霉素(rapamycin)、Mdivi-1以及联合rapamycin和Mdivi-1进行干预,分别从mRNA以及蛋白水平检测自噬相关分子的改变,并同时检测神经元损伤情况。结果 OGD/R处理后,神经元自噬随再灌注时间增长而上调,在OGD 2 h/R 24 h时,Beclin 1以及微管相关轻链蛋白3 II/I(LC3 II/I)比值均达峰值(P<0.001,P<0.01)。细胞活力结果显示,rapamycin可减轻OGD/R损伤(P<0.01),Mdivi-1则会加重OGD/R损伤(P<0.05),但rapamycin可通过增加自噬来减轻Mdivi-1造成的损伤(P<0.05)。蛋白检测结果显示,OGD/R处理后,Mdivi-1可明显降低神经元细胞内Beclin 1和LC3 II的表达(P<0.05)。结论 Mdivi-1可能通过抑制自噬的发生,从而加重神经元OGD/R损伤。Objective The objective of the research is to study the roles and mechanisms of mitochondrial fission inhibitor（ Mdivi-1） in primary cultured cortex neurons of mice after Oxygen-glucose deprivation / Reperfusion（ OGD/R） injury.Methods Occurrence of autophagy was detected after establishing OGD/R model in primary cultured cortex neurons.The mRNA and protein levels of autophagy-related genes were detected accompanied treatment with autophagy inhibitor（ 3-MA）,autophagy agonist（ rapamycin） and Mdivi-1.Moreover,the neural injury was measured.Results Following OGD / R treatment,autophagy was increased dependent on reperfusion times.And both levels of Beclin 1 and Microtubule-associated protein light chain 3 II（ LC3 II） peaked at 24 h（ P〈0.001,P〈0.01）.Results of cell counting kit-8（ CCK-8） after OGD / R treatment showed that Mdivi-1 could aggravate the injury（ P〈0.05）,however,rapamycin could reduce not only the OGD / R injury（ P〈0.01）,but also the aggravating damage induced by Mdivi-1（ P〈0.05）.Furthermore,Mdivi-1 significantly reduced the levels of Beclin 1and LC3 II following OGD / R injury（ P〈0.05）.Conclusion Mdivi-1 could aggravate the injury induced by OGD / R through inhibiting the occurrence of autophagy.

关 键 词：脑缺血再灌注损伤 线粒体分裂 自噬 

分 类 号：R651.15[医药卫生—外科学]