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

摘要

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.%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10滋mol/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，10滋mol/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。