缺氧后胚鼠神经元自噬及凋亡相关蛋白的表达

Changes of LC3 and caspase-3 protein expression in rat fetal cortical neurons afteroxygen deprivation

目的观察缺氧致胚鼠神经元损伤后,凋亡和自噬的发生及变化情况,并探讨其在脑细胞缺血缺氧中的意义。方法取大鼠的胚鼠皮质神经元进行体外原代培养,建立缺氧(oxygen deprivation,OD)损伤模型,通过Western blot方法检测不同缺氧时间点自噬微管相关蛋白轻链3抗体(microtubule-associated protein 1 light chain 3,LC3)以及半胱天冬蛋白酶3(caspase-3)的表达情况。结果神经元于缺氧后可观察到自噬的发生,同时凋亡也随之出现,两者的标志蛋白于OD 0.5h开始表达,并且表达逐渐增加,于OD 4h时间点到达高峰,OD 6h时后凋亡标志蛋白caspase-3继续上调,自噬的标志蛋白LC3表达不再增强。结论 (1)除凋亡外,单纯缺氧能诱导胚鼠神经元发生自噬现象;(2)缺氧所致神经元的损伤过程中,凋亡与自噬同时发生,且表达趋势在OD 4h内一致,OD 4h后细胞凋亡增强。

Objective To make a observation of the variation of autophagy and apoptosis induced by hypoxia in rat fetal cortical neurons,and to lay the foundation for the further relevant investigation.Methods The cerebral cortical neurons of fetal SD rats（within 16～18 days） were cultured to mature and then submitted to oxygen deprivation（OD）.Western blot was utilized for detecting the level of microtubule-associated protein 1 light chain 3（LC3） and caspase-3 conversion.Results Autophagy and apoptosis coexist in the OD process in neurons.LC3 and caspase-3 started to be detected at OD 0.5h,reached the peak at OD 4h and lasted to OD 6h until neurons elapsed.Conclusion Hypoxia is capable of inducing autophagy in rat fetal cortical neurons besides apoptosis.Instead of occurs one by one,autophagy and apoptosis coexist in the stress,and the two phenomenon appear the similar tendency within OD 4h,and apoptosis preponderates over autophagy after OD 4h.