SH-SY5Y细胞中α7尼古丁受体拮抗或基因沉默对Tau蛋白S404和S214位点磷酸化水平的影响及其与p38 MAPK

发布时间：2018-04-09 21:41

本文选题：阿尔茨海默病　切入点：Tau　出处：《贵州医科大学》2017年硕士论文

【摘要】：目的:研究α7尼古丁受体(n ACh R)拮抗或基因沉默对SH-SY5Y细胞Tau蛋白S404和S214位点磷酸化水平的影响及其与p38 MAPK通路的关系,探讨α7 n ACh R调节Tau蛋白位点磷酸化的相关机制。方法:用α7 n ACh R拮抗剂MLA、p38 MAPK通路阻滞剂SB203580及通路激动剂Anisomycin分别或联合处理SH-SY5Y细胞。用Real-time PCR法和Western blotting法分别测定沉默细胞中α7 n ACh R在m RNA和蛋白质表达水平的变化;Western blotting方法测定细胞Tau蛋白、p-Tau(S404)、p-Tau(S214)、p38 MAPK及p-p38 MAPK(Thr180/Tyr182)蛋白水平。结果:α7 n ACh R沉默组与空质粒组相比,α7 n ACh R m RNA和蛋白质水平分别降低了91%(P0.01)和80%(P0.01);p-Tau(S404)蛋白水平升高了71%(P0.01),p-Tau(S214)蛋白水平升高了73%(P0.01),p-p38蛋白水平升高了65%(P0.01)。此外,Anisomycin与MLA、SB203580与MLA、单独MLA处理组共同检测Tau蛋白S404和S214位点磷酸化水平及通路蛋白表达情况,结果显示,MLA处理组和Anisomycin与MLA共同处理组均引起p-Tau(S404)、p-Tau(S214)及p-p38 MAPK蛋白水平明显升高(P0.01),SB203580与MLA共同处理组引起p-Tau(S404)、p-Tau(S214)和p-p38 MAPK蛋白水平显著降低(P0.01)。结论:α7 n ACh R基因沉默或受体拮抗可导致Tau蛋白S404和S214位点磷酸化水平升高并激活p38 MAPK信号通路;抑制p38 MAPK信号转导通路可使MLA诱导的Tau蛋白S404和S214位点磷酸化水平显著降低。
[Abstract]:Aim: to investigate the effects of 伪 7 nicotine receptor n ACh R antagonism or gene silencing on the phosphorylation of S404 and S214 sites of Tau protein in SH-SY5Y cells and its relationship with p38 MAPK pathway, and to explore the mechanism of 伪 7 n ACh R regulating the phosphorylation of Tau protein sites.Methods: SH-SY5Y cells were treated with 伪 7 n ACh R antagonist MLAA p38 MAPK pathway blocker SB203580 and pathway agonist Anisomycin, respectively.The expression of 伪 7 n ACh R in m RNA and protein was measured by Real-time PCR assay and Western blotting method. The protein levels of Tau protein p-Taun S404 and p-p38 MAPKtr 180 / Tyr182 were measured by Real-time PCR and Western blotting.In addition, Anisomycin and MLA-SB203580 and MLA-treated group were used to detect the phosphorylation level of Tau protein S404 and S214 and the expression of pathway protein.Conclusion: 伪 7n ACh R gene silencing or receptor antagonism can increase the phosphorylation level of S404 and S214 sites of Tau protein and activate p38 MAPK signaling pathway, and inhibit the phosphorylation level of Tau protein S404 and S214 sites induced by MLA.
【学位授予单位】：贵州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.16

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