【摘要】 目的:研究Che-1蛋白对氧糖剥夺(Oxygen glucose deprivation, OGD)所致神经元损伤的保护作用及机制。方法:OGD处理神经元后,采用免疫荧光染色和免疫印迹法检测Che-1蛋白的表达;慢病毒转染神经元实现Che-1过表达,检测乳酸脱氢酶(Lactate dehydrogenase, LDH)释放量和流式细胞术检测神经元凋亡反映OGD所致神经元损伤程度,采用免疫荧光染色和免疫印迹法检测神经元自噬;使用自噬激动剂雷帕霉素(Rapamycin)处理神经元,并通过检测LDH释放量和流式细胞术研究自噬在Che-1保护作用中的作用。结果:免疫荧光结果显示,OGD后神经元Che-1蛋白表达明显增高;免疫印迹结果显示,OGD后6至48 h神经元Che-1蛋白表达明显增高;慢病毒转染过表达Che-1蛋白后,OGD所致神经元LDH释放量明显减低,且OGD所致神经元凋亡明显减少;过表达Che-1蛋白可显著减少OGD所致神经元Beclin1和LC3II的表达;自噬激动剂Rapamycin可逆转Che-1对OGD所致神经元损伤的保护作用。结论:过表达Che-1蛋白可通过抑制神经元自噬对OGD所致神经元损伤发挥保护作用。更多还原

【Abstract】 Objective: The aim of the present study was to investigate the protective effect of Che-1 on neuronal injury induced by oxygen glucose deprivation(OGD) and the potential underlying mechanisms. Methods: After treatment with OGD, immunostaining and western blot were performed to detect the expression of Che-1 in neurons. Transfection with lentivirus was used to overexpress Che-1,and lactate dehydrogenase(LDH) release and flow cytometry were performed to measure neuronal injury. Immunostaining and western blot were used to determine autophagy in neurons. After treatment with the autophagy activator rapamycin, LDH release and flow cytometry were performed to investigate the protective effect of Che-1 against OGD. Results: The results of immunostaining showed that OGD significantly increased the expression of Che-1 in cortical neurons. The results of western blot showed that OGD significantly increased the expression of Che-1 from 6 to 48 h. Overexpression of Che-1 alleviated the OGD-induced LDH release and apoptotic neuronal death.Overexpression of Che-1 significantly decreased the expression of Beclin1 and LC3 II in neurons. The autophagy activator rapamycin partially prevented the Che-1-induced protection against OGD in neurons. Conclusions: Overexpression of Che-1 could exert protective effect against OGD-induced neuronal injury through activating autophagy in cortical neurons.更多还原