基础医学与临床 ›› 2015, Vol. 35 ›› Issue (5): 579-584.

• 研究论文 •    下一篇

miR-124-3p通过负调控Caveolin-1表达降低N2a/APPswe细胞内Tau蛋白磷酸化水平

向月1,张雄1,杨军1,戴颂阳2,李昱1   

  1. 1. 重庆医科大学
    2. 重庆医科大学神经科学研究中心
  • 收稿日期:2014-10-08 修回日期:2015-01-01 出版日期:2015-05-05 发布日期:2015-04-28
  • 通讯作者: 李昱 E-mail:liyu100@163.com
  • 基金资助:
    国家自然科学基金资助项目

miR-124-3p inhibits N2a/APPswe cells Tau phosphorylation levels via down-regulating Caveolin-1

  • Received:2014-10-08 Revised:2015-01-01 Online:2015-05-05 Published:2015-04-28

摘要: 探讨在阿尔茨海默病(AD)细胞模型中miR-124-3p通过调控Caveolin-1的表达对细胞内Tau蛋白磷酸化水平的影响。方法 体外培养野生型N2a(N2a/WT)和N2a/APPswe细胞,荧光定量PCR及Western blot分别检测miR-124-3p、APP和Caveolin-1的表达;N2a/APPswe细胞分别转染miR-124-3p模拟物( mimics)、Caveolin-1过表达载体及干扰RNA后,用荧光定量PCR及Western blot分别检测Caveolin-1、Tau和Tau-Ser404表达。 结果 与野生型N2a细胞比较,N2a/APPswe细胞中miR-124-3p表达降低(P<0.01),APP表达升高(P<0.01),Caveolin-1表达升高(P<0.01)。N2a/APPswe细胞转染miR-124-3p mimics后,Caveolin-1表达降低(P<0.01),Tau-Ser404/Tau降低(P<0.01)。N2a/APPswe细胞转染Caveolin-1过表达载体后, Tau-Ser404/Tau升高(P<0.01),转染干扰RNA后Tau-Ser404/Tau降低(P<0.01)。结论 miR-124-3p可能通过调节其靶基因Caveolin-1的表达降低Tau蛋白磷酸化水平在AD中发挥神经保护作用。

关键词: miR124-3p, caveolin-1, N2a/APPswe细胞, Tau蛋白, 磷酸化

Abstract: Objective To explore of the effect of miR-124-3p on the level of Tau phosphorylation through regulating caveolin-1 in Alzheimer,s diseases cell model. Mehods Wild type N2a (N2a/WT) and N2a/APPswe cells were cultured in vitro, real-time fluorescence quantitative PCR (Realtime-PCR) and Western blot(WB)was respectively used to detect the expression levels of miR-124-3p and amyloid precursor protein and caveolin-1; N2a/APPswe cell was respectively transfected by miR-124-3p mimics、caveolin-1 overexpression vector and caveolin-1 siRNA.Realtime-PCR and WB was respectively used to detect the expression levels of caveolin-1 and Tau and Tau-Ser404. Results Compared with Wild type N2a cell, in N2a/APPswe cell the expression of miR-124-3p was decreased (P<0.01), while APP and caveolin-1 was increased (P<0.01). After N2a/APPswe cell was transfected with miR-124-3p mimics, the expression levels of caveolin-1 was decreased (P<0.01) and Tau-Ser404/Tau was decreased (P<0.01);after transfection of caveolin-1 overexpression vector, the expression levels of caveolin-1 was increased (P<0.01) and Tau-Ser404/Tau was increased (P<0.01); after transfection of caveolin-1 siRNA the expression levels of caveolin-1 was decreased (P<0.01) and Tau-Ser404/Tau was decreased (P<0.01). Conclusions miR-124-3p may reduce the level of Tau phosphorylation and play a neuroprotective role in AD through regulating the expression of caveolin-1.

Key words: miR124-3p, caveolin-1, N2a / APPswe cells, Tau protein, phosphorylation

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