您的位置：山东大学 -> 科技期刊社 -> 《山东大学学报（医学版）》

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

PDF (PC)

438

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
[1] Qing H, Zhou W, Christensen MA, et al. Degradation of BACE by the ubiquitin-proteasome pathway[J]. FASEB J, 2004, 18(13):1571-1573.
[2] Chen CH, Zhou W, Liu S, et al. Increased NF-κB signalling up-regulates BACE1 expression and its therapeutic potential in Alzheimer's disease[J]. Int J Neuropsychopharmacol, 2012, 15(1):77-90.
[3] Hirai K, Aliev G, Nunomura A, et al. Mitochondrial abnormalities in Alzheimer's disease[J]. J Neurosci, 2001, 21(9):3017-3023.
[4] Geisler S, Holmstr m KM, Skujat D, et al. PINK1/Parkin-mediated mitophagy is dependent on VDAC1 and p62/SQSTM1[J]. Nat Cell Biol, 2010, 12(2):119-131.
[5] Soldano A, Hassan BA. Beyond pathology: APP, brain development and Alzheimer's disease[J]. Curr Opin Neurobiol, 2014, 27:61-67.
[6] Fakevall A, Alikhani N, Bhushan S, et al. Degradation of the amyloid beta-protein by the nove mitochondrial petidasome, PreP[J]. J Biol Chem, 2006, 281(39):29096-29104.
[7] 李丽, 张杰, 余焕玲,等. β淀粉样肽31-35介导的神经细胞线粒体损伤及机制研究[J]. 中国食品卫生杂志, 2008, 20(2):107-110. LI Li, ZHANG Jie, YU Huanling, et al. Study on damage and mechanism of mitochondria of neurons induced by Aβ31-35[J]. Chinese Journal of Food Hygiene, 2008, 20(2):107-110.
[8] Amadoro G, Corsetti V, Florenzano F, et al. Morphological and bioenergetic demands underlying the mitophagy in post-mitotic neurons: the pink-parkin pathway[J]. Front Aging Neurosci, 2014, 6:1-18.
[9] 刘晓婷, 王延让, 张明. 线粒体介导细胞凋亡的研究进展[J]. 环境与健康杂志, 2013, 30(2):182-184. LIU Xiaoting, WANG Yanrang, ZHANG Ming. Mitochondria-mediated apoptosis: a review of recent studies[J]. Journal of Environment and Health, 2013, 30(2):182-184.
[10] 鲁严, 周梅华. 线粒体自噬和线粒体凋亡及其相关病理生理机制研究进展[J]. 实用老年医学, 2011, 25(1):79-81. LU Yan, ZHOU Meihua. The related pathophysiological mechanism of mitochondrial autophagy and mitochondria apoptosis: a review of recent studies[J]. Pract Geriatr, 2011, 25(1):79-81.
[11] Trempe JF, and Fon EA. Structure and function of parkin, PINK1, and DJ-1, the three musketeers of neuroprotection[J]. Front Neurol, 2013, 4:1-11.
[12] Narendra D, Tanaka A, Suen DF, et al. Parkin is recruited selectively to impaired mitochondria and promotes their autophagy[J]. J Cell Biol, 2008, 183(5):795-803.
[13] Matsuda N, Sato S, Shiba K, et al. PINK1 stabilized by mitochondrial depolarization recruits Parkin to damaged mitochondria and activates latent Parkin for mitophagy[J].J Cell Biol, 2010, 189(2):211-221.
[14] Büeler H. Mitochondrial dynamics, cell death and the pathogenesis of Parkinson's disease[J]. Apoptosis, 2010, 15(11):1336-1353.
[15] 李清华, 肖福英. 线粒体自噬最新研究进展[J]. 国际遗传学杂志, 2012, 35(1):10-20. LI Qinghua, XIAO Fuying. Advances in the regulation of mitophagy[J]. Int J Genet, 2012, 35(1):10-20.
[16] Gómez-Sánchez R, Gegg ME, Bravo-San Pedro JM, et al. Mitochondrial impairment increases FL-PINK1 levels by calcium-dependent gene expression[J]. Neurobiol Dis, 2014, 62:426-440.
[17] 杨辉, 左伋, 刘雯. Parkin、PINK、DJ-1和线粒体功能障碍与帕金森病[J]. 生命科学, 2010, 22(10):1009-1012. YANG Hui, ZUO Ji, LIU Wen. Parkin, PINK1, DJ-1 and mitochondria dysfunction with Parkinson's disease[J]. Chinese Bulletin of Life Sciences, 2010, 22(10):1009-1012.
[18] Kabeya Y, Mizushima N, Ueno T, et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing[J]. EMBO J, 2000, 19(21):5720-5728.
[19] Kadowaki M, Karim MR. Cytosolic LC3 ratio as a quantitative index of macroautophagy[J]. Methods Enzymol, 2009, 452:199-213.
[1] 闫鹏 王蓉 杜怡峰 沈伦乾. 老年性痴呆患者尿中AD7c-NTP含量的研究[J]. 山东大学学报(医学版), 2209, 47(6): 106-.
[2] 李蒙蒙,王苗苗,刁雪琴,田克立,徐霞,任桂杰. 阿尔茨海默病细胞模型中lncRNA RP11-543N12.1对CDH13表达的调控作用[J]. 山东大学学报（医学版）, 2017, 55(3): 12-18.
[3] 唐毅. 阿尔茨海默病诊断标准:从临床诊断到病理生理诊断[J]. 山东大学学报（医学版）, 2017, 55(10): 14-20.
[4] 杜蘅,袁晓东. 阿尔茨海默病病因及发病机制研究进展[J]. 山东大学学报（医学版）, 2017, 55(10): 21-27.
[5] 仇成轩, 杜怡峰. 重视阿尔茨海默病和老年痴呆症的人群干预研究[J]. 山东大学学报（医学版）, 2017, 55(10): 1-6.
[6] 杜怡峰,井然. 阿尔茨海默病的诊断与治疗[J]. 山东大学学报（医学版）, 2017, 55(10): 7-13.
[7] 李懋禹,唐诗,杜怡峰. TRB3在APP/PS1转基因小鼠脑内的表达及其意义[J]. 山东大学学报（医学版）, 2016, 54(10): 1-5.
[8] 陈思, 王牧川, 任楠楠, 许巍, 王玲玲, 栾海辉, 马俊, 刘艺鸣. Parkin基因多态性与山东省汉族帕金森病发病风险的关系[J]. 山东大学学报（医学版）, 2015, 53(4): 71-74.
[9] 葛树建, 林炜炜, 丛琳, 张涛, 韩晓娟, 张清华, 杜怡峰. 基于Web构建山东省阿尔茨海默病临床数据库[J]. 山东大学学报（医学版）, 2015, 53(10): 42-45.
[10] 李梅, 孟庆慧, 蔡巧英, 徐雁, 范晓婷. 乙酰葛根素对Aβ25-35诱导BV-2小胶质细胞Caspase-3表达的影响[J]. 山东大学学报（医学版）, 2015, 53(10): 32-36.
[11] 王金红, 李文涛, 于树田, 董丽. 多奈哌齐对阿尔茨海默病模型大鼠下丘脑-垂体-肾上腺轴的影响[J]. 山东大学学报（医学版）, 2014, 52(7): 11-15.
[12] 钱冰，吕春娥，田晓康，高殿帅，刘志安. 钙调蛋白依赖性蛋白激酶Ⅱ在AD样大鼠海马的表达及其与细胞凋亡的关系[J]. 山东大学学报（医学版）, 2014, 52(2): 44-47.
[13] 亓珅，韩晓娟，王璐，张清华，杜怡峰. 海风藤提取物对激活的小胶质细胞IL-1β和TNF-α表达的影响[J]. 山东大学学报(医学版), 2013, 51(5): 11-14.
[14] 殷青青，刘雪平，董传芳，董雪丽，李艳菊, 罗鼎真，侯训尧. 糖基化终末产物对大鼠基底前脑胆碱能神经元的损伤作用[J]. 山东大学学报(医学版), 2012, 50(8): 20-.
[15] 陈敏，单培彦，刘爱芬，麻琳，于晓琳. 阿尔茨海默病患者血糖、胰岛素及胰岛素降解酶水平的研究[J]. 山东大学学报(医学版), 2012, 50(3): 75-.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 2018 《山东大学学报(医学版)》编辑部　
地址:山东大学科技期刊社（济南市历城区山大南路27号山东大学中心校区明德楼B座721室） 电话: 0531-88366918 E-mail: xbyxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发