【摘要】：目的探討Apo E4與GSK-3β及Tau蛋白超磷酸化之間的關系,為研究Apo E4在阿爾茨海默病(AD)中的作用機制提供實驗依據(jù)。方法分別將對照質(zhì)粒載體p IRES-EGFP、重組質(zhì)粒Apo E4/p IRES-EGFP和Apo E3/p IRES-EGFP轉(zhuǎn)染U87腦星形膠質(zhì)細胞系,用免疫蛋白印跡法(Western blotting)檢測p-Tau/Tau及GSK-3β蛋白水平的變化。在U87細胞中轉(zhuǎn)入干擾Apo E表達的si RNA(Apo E-si RNA)及對照si RNA,檢測p-Tau/Tau及GSK-3β蛋白的改變情況。將質(zhì)粒轉(zhuǎn)染前后及si RNA干擾前后目標蛋白的含量或磷酸化水平的改變進行統(tǒng)計學比較。結(jié)果與對照組相比,GSK-3β蛋白水平的相對值分別為:Apo E4組(1.819±0.130,P0.01,n=3)和Apo E3組(1.336±0.130,P0.01,n=3),差異具顯著差異,且Apo E4和Apo E3組比較也具有顯著性差異(P0.01)。同樣,相對于對照組,Apo E4和Apo E3組的磷酸化Tau蛋白(p-Tau)的相對值分別為1.587±0.027(P0.01,n=3)和1.436±0.026(P0.01,n=3),均具顯著差異;且Apo E4組較Apo E3組的促進作用更強,兩組間具顯著差異(P0.01)。此外,用Apo E-si RNA干擾后,U87細胞中GSK-3β的表達和p-Tau水平均較對照組顯著降低,分別為0.544±0.058(P0.001,n=3)和0.474±0.060(P0.01,n=3)。結(jié)論 AD危險因子Apo E4可能通過上調(diào)GSK-3β的表達而促進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 鹵0.130, P0.01, n 鈮，

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