摘要
目的:研究神经毒性物质β-淀粉样肽(β-AP)?N-甲基-D-天冬氨酸(NMDA)对原代培养鼠脑皮层神经细胞的致凋亡损伤和维生素E可能的神经保护作用?方法:通过形态学观察和MTT自动比色法检测存活细胞率;用DNA电泳和流式细胞技术分析细胞凋亡率;用免疫细胞化学技术分析细胞凋亡相关基因表达?结果:经β-AP?NMDA作用后,培养神经细胞出现明显的细胞凋亡特征:存活细胞减少,凋亡细胞数增多及特征性DNA断裂带,神经细胞凋亡相关蛋白Bcl-2表达下降?Bax表达上升(均P<0.05)?加维生素E组细胞则表现以上凋亡特征减弱?结论:神经毒性物质β-AP?NMDA可诱导培养神经细胞凋亡,细胞凋亡机制与氧化损伤相关,维生素E可减缓细胞凋亡?
Objective: To study apoptosis of the cultured rat brain neurons induced by β-amyloid peptide(β-AP) and N-methyl-D-aspartate (NMDA) and neuro-protective effect of vitamin E. Methods: The morphological observation and MTT method were used to assay cell survival. Neuron apoptosis were assayed with DNA agarose gel electrophoresis and the flow cytometric analysis, and the expression of apoptosis relative genes by immunocytochemistry staining method. Results: After treated with β-AP or NMDA, the neurons showed clear apoptotic characteristics: the survival cells in MTT assay decreased, the DNA cleavage fragment bands in electrophoresis were detected, and the apoptosis cell rate increased in above assays. In immunocytochemistry apoptotic protein assay, the expression of Bcl-2 was lower and expression of Bax higher significantly in β-AP or NMDA groups(P<0.05). But the above apoptotic characteristics decreased in vitamin E added group. Conclusion: β-AP or NMDA can induce cultured neurocyte apoptosis, the mechanism of which possibley relates to oxidative impairment. The vitamin E may attenuate that effect. [
出处
《山东大学学报（医学版）》
CAS
2004年第3期265-268,273,共5页
Journal of Shandong University:Health Sciences
基金
山东省卫生厅九五攻关课题
