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

发布时间：2018-08-07 21:40

【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%(P0.01)和80%(P0.01);tau蛋白水平升高了79%(P0.01);p-tau(S404)蛋白水平升高了74%(P0.01);p-tau(S214)蛋白水平升高了72%(P0.01);p38蛋白水平升高了64%(P0.01);p-p38蛋白水平升高了67%(P0.01)。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。
[Abstract]:Objective to investigate the effects of 伪 7 nicotine receptor (nAChR) silencing on the level of tau protein phosphorylation and p38 MAPK pathway in SH-SY5Y cells, and to explore the regulatory effect of 伪 7 nAChR on tau protein phosphorylation and the relationship between 伪 7 nAChR and p38 MAPK pathway. Methods the expressions of 伪 7 nAChR in mRNA and protein in 伪 7 nAChR silencing cells were measured by Real-time PCR and Western blotting respectively. The protein levels of tau protein p-tau (S404) -p-tau (S214) p38 MAPK and p-p38 MAPK (Thr180/Tyr182) were detected by Western blotting. Results compared with the blank plasmid group, 伪 7 nAChR mRNA and protein levels in 伪 7nAChR silencing group decreased by 91% (P0.01) and 80% (P0.01) respectively, the protein level of p-tau (S404) increased by 79% (P0.01), the protein level of p-tau (S214) increased by 74% (P0.01), the protein level of p-tau (S214) increased by 72% (P0.01) and the protein level of p-p38 increased by 64% (P0.01) by 67% (P0.01). Conclusion 伪 7 nAChR silencing may lead to excessive phosphorylation of tau protein, which may be related to the activation of p38 MAPK pathway.
【作者单位】：贵州医科大学分子生物学重点实验室;
【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]
【分类号】：R749.16

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