目的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、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模拟物后,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TauSer404/Tau降低(P〈0.01)。结论 miR-124-3p可能通过调节其靶基因Caveolin^-1的表达而降低Tau蛋白磷酸化水平,在AD中发挥神经保护作用。
Objective To explore the effect of miR-124-3p on Tau phosphorylation level through regulating Caveolin-1 in Alzheimer's diseases( AD) cell model. Methods Wild type N2a( N2 a / WT) and N2 a / APPswe cells were cultured in vitro. Real-time fluorescence quantitative PCR( real time-PCR) and Western blot were respectively used to detect the expression level of miR-124-3p,amyloid precursor protein and Caveolin-1; N2 a / APPswe cells were respectively transfected by miR-124-3p mimics,Caveolin-1 overexpression vector and Caveolin-1 siRNA. Real time PCR and Western blot were respectively used to detect the expression levels of Caveolin-1,Tau and TauSer404. Results Compared with wild type N2 a cell,the expression of miR-124-3p in N2 a / APPswe cell was decreased( P〈0. 01),while APP and Caveolin-1 were increased( P〈0. 01). After N2 a / APPswe cells were transfected with miR-124-3p mimics,the expression level 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 level of Caveolin-1was increased( P〈0. 01) and Tau-Ser404 / Tau was increased( P〈0. 01); after transfection of Caveolin-1 siRNAthe expression level 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.