Aβ和氧自由基诱导的培养神经元凋亡及维生素E的保护作用

The induction of cultured neurons apoptosis by β-amyloid peptide and oxygen-derived free radicals and protective role of vitamin E

目的研究β淀粉样肽(Aβ)、反应活性氧(ROS)对原代培养鼠脑皮层神经细胞凋亡的诱导及维生素E(VE)可能的保护作用。方法通过形态学观察、MTT法检测、流式细胞技术分析、DNA琼脂糖凝胶电泳、免疫细胞化学技术研究Aβ、ROS对培养皮层神经细胞的损伤。结果经Aβ、活性氧的作用,神经细胞出现明显的凋亡特征,OD值降低,DNA电泳显示出现断裂DNA片段。凋亡细胞数增多。免疫细胞化学显示Bcl2表达下降,Bax表达增加(均P<0.05),而VE组可明显改变上述凋亡特征。结论Aβ、活性氧诱导培养皮层神经元凋亡机制可能与氧化损伤有关,维生素E能减缓这一作用。

Objective To study the cultured neurons apoptosis induced by β-amyloid peptide (Aβ) or reactive oxygen species (ROS) and the possible protective effect of vitamin E. Methods The damage of Aβ and ROS on cultured cortical neurons was studied with morphological observation, MTT method assay, flow cytometric analysis, DNA agarose gel electrophoresis and immunocytochemistry staining method. Results After treated with Aβ or ROS, the neurons showed obvious apoptotic characteristics, including decreased OD570 value in MTT method assay, DNA cleavage fragment in electrophoresis and increased apoptotic cells. Immunocytochemistry showed that expression of Bcl-2 reduced and expression of Bax increased significantly (all P<0.05). But the apoptosis characteristics the above were significantly improved in vitamin E group.Conclusions Vitamin E could attenuate the Aβ or ROS-induced neurocyte apoptosis, the mechanism of which may be related to oxidative impairment.