糖元合酶激酶-3过度激活对神经细丝磷酸化的影响  

Effect of glycogen synthase kinase-3 overactivation on neurofilament phosphorylation

作　　者：陈娟[1] 周洁[1] 王建枝[2] 冯友梅[1] 

机构地区：[1]华中科技大学同济医学院基础医学院生物化学与分子生物学系 [2]华中科技大学同济医学院病理生理学系,武汉430030

出　　处：《解剖学杂志》2005年第3期261-263,F002,共4页Chinese Journal of Anatomy

基　　金：国家自然科学基金(30100057;30170221)

摘　　要：目的:在细胞水平研究糖元合酶激酶(GSK-3)过度激活对神经细丝磷酸化的影响。方法:采用磷酯酰肌醇三磷酸激酶(PI3K)的特异性抑制剂渥曼青霉素(wortmannin,WT)处理野生型小鼠成神经瘤细胞株(N2a/wt),免疫印迹技术及酶活性测定检测GSK-3活性,免疫荧光技术检测神经细丝(NF)的磷酸化状态。进一步采用GSK-3特异性的抑制剂LiCl处理上述细胞,检测上述相同指标。结果:WT处理N2a细胞1h后,GSK-3酶活性显著增加,并使抗体SMI31显色增强,SMI32显色减弱,提示NF被过度磷酸化。而采用WT和LiCl联合处理N2a/wt细胞,GSK3活性显著降低,与此同时,NF的磷酸化程度明显减少。结论:GSK3过度激活可引起细胞骨架蛋白NF的异常过度磷酸化,而过度磷酸化的NF可能参与了AD的病理过程。Objective: To investigate the effect of glycogen synthase kinase-3 (GSK-3) overactivation on neurofilament phosphorylation in cultured cells. Methods: After N2a cells were treated with wortmannin or with wortmannin and LiCl, the GSK-3 activity and neurofilament phosphorylation were detected by GSK-3 activity assay, Western blots and immunofluoresence. Results: It was found that overactivation of GSK-3 induced hyperphosphorylation of neurofilament. Overactivation of GSK-3 led to a reduced staining with antibody SMI32 and an enhanced staining with antibody SMI31. Conclusion: These results indicate that in vitro over activation of GSK-3 may lead to neurofilament hyperphosphorylation and this may be the underlying mechanism of Alzheimer's disease.

关 键 词：神经细丝 过度激活 合酶 糖元 GSK-3 异常过度磷酸化 细胞水平研究 特异性抑制剂 免疫印迹技术 细胞骨架蛋白 LiCl 渥曼青霉素 磷酯酰肌醇 酶活性测定 N2A细胞 磷酸激酶 瘤细胞株 技术检测 免疫荧光 联合处理 病理过程 

分 类 号：R749.16[医药卫生—神经病学与精神病学] R363[医药卫生—临床医学]