转染NEP基因对Aβ25-35诱导神经元凋亡的保护作用

山东大学学报(医学版)

转染NEP基因对Aβ_25-35诱导神经元凋亡的保护作用

张亚伦，丁岩，邵世滨，杨玲玲，任凯，段珊，曾季平，崔行

山东大学医学院生物化学与分子生物学研究所，济南 250012

收稿日期:2007-04-26 修回日期:1900-01-01 出版日期:2008-01-16 发布日期:2008-01-16
通讯作者: 崔行

Neuroprotective effects of Neprilysin on neuronal apoptosis induced by Amyloid-beta peptide

ZHANG Ya-lun, DING Yan, SHAO Shi-bin, YANG Ling-ling, REN Kai， DUAN Shan, ZENG Ji-ping, CUI Xing

Institute of Biochemistry and Molecular Biology, School of Medicine, Shandong University, Jinan 250012, China

Received:2007-04-26 Revised:1900-01-01 Online:2008-01-16 Published:2008-01-16
Contact: CUI Xing

摘要/Abstract

摘要： 目的研究转染NEP基因对神经毒物质Aβ_25-35诱导神经元凋亡的保护作用。方法培养大鼠脑皮层神经元，用Aβ_25-35作用神经元制备凋亡模型，NEP基因转染神经元。通过MTT法及流式细胞分析法观察NEP基因过表达对Aβ诱导神经元凋亡的保护作用。通过RT-PCR检测目的基因NEP，APP,以及凋亡相关基因Bcl-2和Bax-mRNA的表达，Western-Blot测定APP和Aβ的蛋白表达。结果MTT和流式细胞分析法观察，发现在Aβ_25-35作用下转染NEP组神经元活,性显著增高(P＜0.05)，凋亡率降低。RT-PCR及Western Blot结果显示，转染NEP组神经元APP基因的表达明显减少，Aβ的生成减少；同时促凋亡基因Bax表达降低，Bcl-2基因表达增高。结论转染NEP基因能保护Aβ所致的神经元凋亡。

Abstract: To explore the protective effects of transfection of the Neprilysin gene on primary cells against apoptosis induced by Aβ_25-35. MethodsPrimary cortical neurons of fetal Wistar rats in vitro were cultured and the apoptosis cell model was induced by Aβ_25-35. The NEP genes were transiently transfected into neurons. Changes of cell apoptosis were determined by MTT assay and flow cytometric DNA analysis. The expression of target mRNA and proteins was determined by RT-PCR and Western blot. ResultsExposure of primary neurons to Aβ_25-35 resulted in changes of cell apoptosis. The MTT assay showed that cell activity remarkably decreased (P＜0.01) and cytometric DNA analysis showed an increase in the apoptosis rate. While in the neurons transfected with NEP genes, there were no such remarkable changes when treated with Aβ_25-35. The suppression effect of the NEP gene on APP expression was shown by RT-PCR and Western blot, and the expression of Bax was decreased while that of Bcl-xl was increased after transfection. ConclusionTransfection of the NEP gene can protect primary neurons from apoptosis induced by Aβ_25-35.

Key words: Neurons, Alzheimer Disease, primary culture, Neprilysin/Neutral endopeptidase

中图分类号:

Q789

张亚伦,丁岩,邵世滨,杨玲玲,任凯,段珊,曾季平,崔行. 转染NEP基因对Aβ_25-35诱导神经元凋亡的保护作用[J]. 山东大学学报(医学版), 2008, 46(1): 5-8.

ZHANG Ya-lun,DING Yan,SHAO Shi-bin,YANG Ling-ling,REN Kai,DUAN Shan,ZENG Ji-ping,CUI Xing. Neuroprotective effects of Neprilysin on neuronal apoptosis induced by Amyloid-beta peptide[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2008, 46(1): 5-8.

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