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  

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

作　　者：向月[1] 张雄[1] 杨军[1] 戴颂阳 李昱[1] 

机构地区：[1]重庆医科大学病理教研室神经科学研究中心神经生物学重点实验室,重庆400016

出　　处：《基础医学与临床》2015年第5期579-584,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81271426)

摘　　要：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.

关 键 词：miR124-3p CAVEOLIN-1 N2a/APPswe细胞 TAU蛋白 磷酸化 

分 类 号：R749.16[医药卫生—神经病学与精神病学]