体外培养皮层神经元缺糖氧后氧化-抗氧化状态的变化

目的:观察皮层神经元缺糖氧复氧后氧化-抗氧化分子的变化,并探讨其对凋亡的影响.方法:建立体外培养皮层神经元缺糖氧复氧损伤模型,实验分为正常对照组,缺糖氧复氧不同时间组(3、6、12及24 h组),检测各组细胞上清液乳酸脱氢酶(LDH)、一氧化氮(NO)含量及一氧化氮合酶(NOS)活性;观察神经元谷胱甘肽(GSH)含量及谷胱甘肽过氧化物酶(GPx)活性变化:Hochest33258核染色观察皮层神经元凋亡情况.结果:与正常对照组相比,缺糖氧复氧后神经元LDH释放量及NO含量明显增多、GPx活性持续下降(P<0.05);于缺糖氧复氧3 h及6 h时,NOS活性增高、GSH含量减少(P<0.05),而于24 h时二者变化无显著差异(P>0.05);神经元于缺糖氧复氧3 h时出现凋亡,且随复氧时间的延长,凋亡持续增多(P<0.05).结论:随着缺糖氧复氧时间的持续,氧化及抗氧化平衡的破坏,可能是致体外培养皮层神经元凋亡的原因之一.

The oxidant-antioxdant change of cortical neurons to anoxia-reoxygen injury in vitro

Objective:To investigate the effect of apoptosis of oxidant-antioxidant change on anoxia-reoxygen injury to cortical neurons in vitro.Methods:Acute glucose-oxygen deprivation and subsequent reoxygenation were used to establish anoxia-reoxygenation injury model in cultured cortical neurons.They were randomly divided into 2 groups:control group,groups with different reoxygenation time(3 h,6 h,12 h and 24 h).The content of dehydrogenase(LDH),glutathione(GSH) and NO,the activity of NOS and glutathion peroxidase(GPx) were measured by assay kits.The morphology of apoptotic cells was observed by Hochest33258 staining.Results:Compared with the control group,the level of LDH,NO and cell apoptotic rate was obviously increased,while the activity of GPx was decreased in reoxygenation groups(P < 0.05);The activity of NOS was obviously increased and the GSH level was decreased at reoxygenation 3 h and 6 h(P < 0.05),but there was no significant difference at reoxygenation 24 h(P > 0.05).Conclusion:The imbalance between the system of oxidation and reduction may be one of the reasons inducing neurons apoptosis as reoxygenation time extending in vitro.