Mdivi-1对糖氧剥夺后神经元凋亡蛋白Bax活化、嵌入及细胞色素C释放的影响

目的:研究线粒体分裂蛋白抑制剂Mdivi-1对糖氧剥夺(OGD)后神经元凋亡蛋白Bax活化、嵌入及细胞色素C(Cyt C)释放的影响。方法:体外培养原代皮质神经元并制备OGD模型,采用不同浓度的Mdivi-1干预细胞,MTT测定神经元存活率。之后采用10μmol/L Mdivi-1干预细胞,免疫共沉淀或细胞碱处理后测定细胞凋亡蛋白Bax的活化和嵌入情况。抽提线粒体和细胞浆蛋白,Western Blot测定线粒体内Cyt C释放情况。结果:Mdivi-1可以剂量依赖地改善OGD后神经细胞的存活,10μmol/L Mdivi-1干预可以减少凋亡蛋白Bax的激活和嵌入,并且减少Cyt C的释放。结论:线粒体分裂蛋白抑制剂Mdivi-1在OGD后具有明确的神经保护作用,其机制可能和其抑制细胞凋亡有关。

Mdivi-1 Mediates Apoptotic Protein Bax Insertion Activation and Cytochrome C Release on Oxygen and Glucose Deprived Cortical Neurons

ObjectiveTo study the effect of mitochondrial fission protein inhibitor Mdivi-1 on apoptotic protein Bax activation insertion and cytochrome C release of oxygen and glucose deprived cortical neurons. Methods：Primary cultured cortical neurons were treated with different doses of Mdivi-1 after oxygen-glucose deprivation procedure. MTT was used to evaluate the cell viability. After OGD treatment the neurons were treated with 10 μmol/L Mdivi-1, co-immunoprecipitation or alkali treatment were used to determine Bax activation and insertion. Mitochondrial and cytosolic fraction proteins were extracted, Western Blot was used to detect the cytochrome C release. Results： Mdivi-1 significantly improved cell viability in a dose-dependent manner. Mdivi-1 blocked apoptotic protein Bax insertion and activation and subsequent cytochrome C release induced by OGD. Conclusion： Mitochondrial fission protein inhibitor Mdivi-1 has a neuroprotective effect on neurons after OGD, probably mediated by anti-apoptotic effects.