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抑制聚ADP-核糖聚合酶-1活性降低HEK293/tau441细胞tau蛋白的磷酸化

陈钦浩, 廖晓梅, 王少辉^*

华中师范大学生命科学学院，武汉 430079

摘要

为了研究聚ADP-核糖聚合酶-1 [poly(ADP-ribose)polymerase-1, PARP-1]对微管相关蛋白tau磷酸化水平的影响，本实验分别用不同剂量(0.5, 1, 2, 4 mmol/L)的PARP-1的抑制剂3-氨基苯甲酰胺(3-aminobenzamide, 3-AB)处理稳定表达tau441蛋白的HEK293细胞。24 h后观察细胞的形态学变化，并用免疫印迹的方法检测PARP-1的聚ADP-核糖化修饰变化、微管相关蛋白tau的磷酸化水平和糖原合酶激酶-3 (glycogen synthase kinase 3, GSK-3)的活性改变情况。结果显示：(1)不同浓度的3-AB处理HEK293/tau441细胞后，细胞形态未发生显著的变化；(2) PARP-1的活性下降引起tau蛋白Ser195/198/199/202位点的非磷酸化增强；(3) PARP-1的活性抑制使tau蛋白Thr231位点的磷酸化下降，同时细胞内的GSK-3的活性也下降。结果提示，PARP-1的活性抑制可能通过下调GSK-3的活性，降低tau蛋白磷酸化水平。

关键词： 聚ADP-核糖聚合酶-1; 糖原合酶激酶-3; Tau蛋白; 阿尔茨海默病

分类号：R363

[Suppressing poly(ADP-ribose)polymerase-1 inhibits tau phosphorylation in HEK293/tau441 cells.] [Article in Chinese]

CHEN Qin-Hao, LIAO Xiao-Mei, WANG Shao-Hui^*

College of Life Science, Huazhong Normal University, Wuhan 430079, China

Abstract

The study aimed to investigate the effect of inhibition of poly(ADP-ribose) polymerase-1 (PARP-1) activity on tau phosphorylation in HEK293/tau441 cells and its mechanism. HEK293/tau441 cells were treated with 3-aminobenzamide (3-AB), a PARP-1 inhibitor, at different doses (0.5, 1, 2, 4 mmol/L). After 24 h, the cell morphology was observed under phase contrast microscope, tau phosphorylation level in different sites (tau-1, tau-5, Thr231) and the activity of glycogen synthase kinase 3 (GSK-3) were detected by Western blotting. The results showed: (1) 3-AB at different doses failed to change the morphology of cells; (2) The 3-AB-induced decrease in activity of PARP-1 resulted in increase of unphosphorylation level in tau-1(Ser195/198/199/202) sites; (3) The phosphorylation of tau was decreased in Thr231 site, while the total tau was slightly changed after 3-AB treatment; (4) With the increased phosphorylation of GSK-3 at Ser9 site, the activity of GSK-3 was decreased after 3-AB treatment. The results suggest that the inhibition of PARP-1 by 3-AB could decrease tau phosphorylation in HEK293/tau441 cells probably through decreasing GSK-3 activity.

Key words: poly(ADP-ribose)polymerase-1; glycogen synthase kinase 3; Tau protein; Alzheimer's disease

收稿日期：2011-04-21　　录用日期：2011-06-16

通讯作者：王少辉　　E-mail: wsh2002505@163.com

引用本文：

陈钦浩, 廖晓梅, 王少辉. 抑制聚ADP-核糖聚合酶-1活性降低HEK293/tau441细胞tau蛋白的磷酸化[J]. 生理学报 2011; 63 (6): 511-516.

CHEN Qin-Hao, LIAO Xiao-Mei, WANG Shao-Hui. [Suppressing poly(ADP-ribose)polymerase-1 inhibits tau phosphorylation in HEK293/tau441 cells.] [Article in Chinese]. Acta Physiol Sin 2011; 63 (6): 511-516 (in Chinese with English abstract).