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山东大学学报（医学版） ›› 2015, Vol. 53 ›› Issue (10): 1-5.doi: 10.6040/j.issn.1671-7554.0.2014.684

• 基础医学 •    下一篇

线粒体自噬在阿尔茨海默病细胞模型中的作用机制

赵雪莲1, 于君2, 谢兆宏1, 曹彦军1, 刘震1, 王晓1, 徐琳琳1, 杨慧1, 郑晓磊1, 沈阳1, 毕建忠1   

  1. 1. 山东大学第二医院神经内科, 山东 济南 250033;
    2. 山东中医药大学附属医院, 山东 济南 250011
  • 收稿日期:2014-10-10 出版日期:2015-10-10 发布日期:2015-10-10
  • 通讯作者: 毕建忠。E-mail:bjz@sdu.edu.cn E-mail:bjz@sdu.edu.cn
  • 基金资助:
    国家自然科学基金(81371420,81171214,81401052);山东省自然科学基金(ZR2011HM064);山东省卫生厅科研项目(2013WS0248)

Mechanism of mitophagy in a cell model of Alzheimer's disease

ZHAO Xuelian1, YU Jun2, XIE Zhaohong1, CAO Yanjun1, LIU Zhen1, WANG Xiao1, XU Linlin1, YANG Hui1, ZHENG Xiaolei1, SHEN Yang1, BI Jianzhong1   

  1. 1. Department of Neurology, the Second Hospital of Shandong University, Jinan 250033, Shandong, China;
    2. The Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan 250011, Shandong, China
  • Received:2014-10-10 Online:2015-10-10 Published:2015-10-10

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摘要： 目的 研究阿尔茨海默病细胞模型20E2自噬情况及其可能机制。方法 应用ELISA检测体外培养的HEK293细胞和20E2细胞(稳定转染APPSW的HEK293细胞)上清Aβ1-40水平、Western blotting检测APP蛋白表达水平,确定20E2细胞是否建模成功。电镜下观察细胞内线粒体,JC-1检测两种细胞线粒体膜电位,流式细胞仪检测细胞凋亡情况。Western blotting检测LC3-II、PINK1、Parkin表达水平。结果 与HEK293细胞相比,20E2细胞APP蛋白、Aβ1-40表达增加(P<0.05),线粒体肿胀、嵴消失、空泡化明显,线粒体膜电位下降,PINK1、Parkin、自噬相关蛋白LC3-II表达增加(P<0.05)。结论 阿尔茨海默病细胞模型20E2线粒体形态改变明显、膜电位下降,这些改变可能通过PINK1、Parkin途径引起线粒体自噬增加。

关键词: 线粒体自噬, PINK1, Parkin, 阿尔茨海默病, 线粒体膜电位

Abstract: Objective To investigate the effect of autophagy involved in Alzheimer's disease cell model 20E2 and its possible mechanism. Methods To determine whether the 20E2 cells model was successfully established, we detected the levels of Aβ1-40 in HEK293 cells and 20E2 cells (HEK293 cells stably expressing Swedish mutant APP) cultured in vitro by ELISA kit, and the expression of APP protein level was detected by Western blotting. The mitochondria in cells was observed by electron microscope. The mitochondrial membrane potential of both cells was detected by fluorescence probe JC-1. Flow cytometry was used to measure the apoptotic rate. LC3-II, PINK1 and Parkin were detected by Western blotting. Results The expression levels of APP protein and Aβ1-40 increased in 20E2 cells compared with those in HEK293 cells. Mitochondrial swollen, cristae disappeared and vacuolization was obviously observed. Mitochondrial membrane potential decreased. The expression levels of PINK1, Parkin and LC3-II increased (P<0.05). Conclusion In Alzheimer's disease cell model 20E2, the mitochondrial morphology changed obviously and membrane potential of mitochondria declined, and these changes may cause the increase of mitochondrial autophagy through PINK1 and Parkin pathway.

Key words: Mitochondrial membrane potential, Parkin, Alzheimer's disease, PINK1, Mitophagy

中图分类号: 

  • R741.02
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