SH-SY5Y细胞α7尼古丁受体基因沉默对tau蛋白磷酸化水平及p38 MAPK信号通路的影响  

Influence of inhibited α7 nicotinic acetylcholine receptor gene expression on tau phosphorylation and p38 MAPK signal transduction pathway in SH-SY5Y cells

作　　者：周飞[1] 吴昌学[1] 李毅[1] 官志忠[1] 齐晓岚[1] 

机构地区：[1]贵州医科大学分子生物学重点实验室,贵州贵阳550004

出　　处：《中风与神经疾病杂志》2016年第8期676-679,共4页Journal of Apoplexy and Nervous Diseases

基　　金：国家自然科学基金资助项目(No.81360178);教育部"长江学者和创新团队发展计划资助"(No.IRT13058);贵州省科技厅重大专项[黔科合重大专项字2014(6008)];贵州省科技厅项目[201344;黔科合SY字(2013)3020]

摘　　要：目的研究α7尼古丁受体(nAChR)沉默对SH-SY5Y细胞tau蛋白磷酸化水平和p38 MAPK通路的影响,探讨α7 nAChR对tau蛋白磷酸化的调节作用及其与p38 MAPK通路的关系。方法 Real-time PCR法和Western blotting法分别测定α7 nAChR沉默细胞中α7 nAChR在mRNA和蛋白表达水平的变化;Western blotting法检测细胞tau蛋白、p-tau(S404)、p-tau(S214)、p38 MAPK和p-p38 MAPK(Thr180/Tyr182)蛋白水平。结果α7nAChR沉默组与空质粒组相比,α7 nAChR mRNA和蛋白质水平分别降低了91%(P<0.01)和80%(P<0.01);tau蛋白水平升高了79%(P<0.01);p-tau(S404)蛋白水平升高了74%(P<0.01);p-tau(S214)蛋白水平升高了72%(P<0.01);p38蛋白水平升高了64%(P<0.01);p-p38蛋白水平升高了67%(P<0.01)。结论α7 nAChR沉默可导致tau蛋白过度磷酸化,这可能和激动p38 MAPK通路有一定关系。Objective To investigate the influence of inhibited gene expression of a l neuronal nicotinic acetylcholinereceptor （nAChR） on tau phosphorylation and p38 MAPK signal transduction pathway in SH-SY5Y cells,to study theeffect of a l nAChR on tau phosphorylation, and the realationship with p38 MAPK transduction pathway. Methods The levelof α7 nAChR mRNA and protein were detected by real-time PCR and Western blotting, respectively. The protein levels oftau,p-tau （S404） , p-tau （S214） ,p38 MAPK and P-p38 MAPK （ Thr180/Tyr182） were determined by Western blotting.Results As compared with negative controls, the expression of a l nAChR at mRNA and protein levels in the SH-SY5Ycells with a l nAChR silencing were decreased by 91% and 80% Respectively. And the expression of tau,p-tau （S404） ,p-tau（S214） ,p38 MAPK and P-p38 MAPK （Thrl80/Tyrl82） proteins were increased by 79% 、74% 、72% 、64%、67% , respectively.Conclusions The inhibited gene expression of α7 nAChR by RNA interference can lead to tau hyperphosphorylation,which may associate to p38 MAPK transduction pathway.

关 键 词：阿尔茨海默病 TAU alnAChR P38MAPK 

分 类 号：R749.1[医药卫生—神经病学与精神病学]