ApoE4在U87细胞中可增加GSK-3β的表达及Tau的磷酸化

发布时间：2019-06-02 05:07

【摘要】：目的探讨Apo E4与GSK-3β及Tau蛋白超磷酸化之间的关系,为研究Apo E4在阿尔茨海默病(AD)中的作用机制提供实验依据。方法分别将对照质粒载体p IRES-EGFP、重组质粒Apo E4/p IRES-EGFP和Apo E3/p IRES-EGFP转染U87脑星形胶质细胞系,用免疫蛋白印迹法(Western blotting)检测p-Tau/Tau及GSK-3β蛋白水平的变化。在U87细胞中转入干扰Apo E表达的si RNA(Apo E-si RNA)及对照si RNA,检测p-Tau/Tau及GSK-3β蛋白的改变情况。将质粒转染前后及si RNA干扰前后目标蛋白的含量或磷酸化水平的改变进行统计学比较。结果与对照组相比,GSK-3β蛋白水平的相对值分别为:Apo E4组(1.819±0.130,P0.01,n=3)和Apo E3组(1.336±0.130,P0.01,n=3),差异具显著差异,且Apo E4和Apo E3组比较也具有显著性差异(P0.01)。同样,相对于对照组,Apo E4和Apo E3组的磷酸化Tau蛋白(p-Tau)的相对值分别为1.587±0.027(P0.01,n=3)和1.436±0.026(P0.01,n=3),均具显著差异;且Apo E4组较Apo E3组的促进作用更强,两组间具显著差异(P0.01)。此外,用Apo E-si RNA干扰后,U87细胞中GSK-3β的表达和p-Tau水平均较对照组显著降低,分别为0.544±0.058(P0.001,n=3)和0.474±0.060(P0.01,n=3)。结论 AD危险因子Apo E4可能通过上调GSK-3β的表达而促进Tau的磷酸化。
[Abstract]:Objective to investigate the relationship between Apo E4 and GSK-3 尾 and Tau protein hyperphosphate acidizing, and to provide experimental basis for the study of the mechanism of Apo E4 in Alzheimer's disease (AD). Methods the control plasmid vector p IRES-EGFP, recombinant plasmid Apo E4 IRES-EGFP and Apo E3 IRES-EGFP were transfected into U87 brain astrocytes cell line, respectively. The changes of p-Tau/Tau and GSK-3 尾 protein levels were detected by immunoblotting (Western blotting). Si RNA (Apo E-si RNA, which interfered with the expression of Apo E, was transferred into U87 cells and the changes of p-Tau/Tau and GSK-3 尾 proteins were detected by si RNA,. The changes of target protein content or phosphorylation level before and after plasmid transfer and si RNA interference were compared statistically. Results compared with the control group, the relative values of GSK-3 尾 protein levels in Apo E4 group (1.819 卤0.130, P0.01, n 鈮，

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