体外培养皮层神经元缺糖氧后氧化-抗氧化状态的变化
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国家自然科学基金资助(30571555)


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

    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-

    Abstract:

    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.

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张劲松,邵斌霞,何 斌,蒋 雷,程 洁,高 蓉,肖 杭.体外培养皮层神经元缺糖氧后氧化-抗氧化状态的变化[J].南京医科大学学报(自然科学版),2008,28(9):1095-1098

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