转染NEP基因对Aβ_(25-35)诱导神经元凋亡的保护作用  

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

作　　者：张亚伦[1] 丁岩[1] 邵世滨[1] 杨玲玲[1] 任凯[1] 段珊[1] 曾季平[1] 崔行[1] 

机构地区：[1]山东大学医学院生物化学与分子生物学研究所,济南250012

出　　处：《山东大学学报（医学版）》2008年第1期5-8,14,共5页Journal of Shandong University:Health Sciences

基　　金：高等学校博士点科研基金(20050422045)

摘　　要：目的研究转染NEP基因对神经毒物质Aβ25-35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Aβ25-35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Aβ25-35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。Objective To explore the protective effects of transfection of the Neprilysin gene on primary cells against apoptosis induced by Aβ25-35 Methods Primary cortical neurons of fetal Wistar rats in vitro were cultured and the apoptosis cell model was induced by Aβ25-35. The NEP genes were transiently transfected into neurons. Changes of cell apoptosis were determined by MTF assay and flow cytometric DNA analysis. The expression of target mRNA and proteins was determined by RT-PCR and Western blot. Results Exposure of primary neurons to Aβ25-35 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 Aβ25-35 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. Conclusion Transfection of the NEP gene can protect primary neurons from apoptosis induced by Aβ25-35.

关 键 词：神经元 阿尔茨海默病 原代培养 中性内肽酶 

分 类 号：Q789[生物学—分子生物学]