姜黄素通过抑制GSK-3β的活性防治AD的体外研究  

Curcumin inhibits the activity of GSK-3β to prevent AD in vitro

作　　者：张雄[1] 李昱[1,2] 

机构地区：[1]重庆医科大学神经科学研究中心,重庆400016 [2]重庆医科大学病理学教研室,重庆400016

出　　处：《中国药理学通报》2009年第11期1507-1512,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30600196);重庆市自然科学基金资助项目(NoCSTC;2006BB5042);2007年教育部留学回国人员科研启动基金(No教外司留[2007]1108号)

摘　　要：目的探讨姜黄素(Curcumin)通过调节糖原合成激酶-3β(GSK-3β)的活性防治AD的机制。方法体外培养SHSY5Y细胞,转染pBACE1-mychis和pAPPswe,姜黄素处理细胞后,通过RT-PCR和Westernblot分别检测GSK-3β、β-链蛋白(β-catenin)和核内转录因子CyclinD1 mRNA水平和蛋白水平的表达情况,免疫荧光法检测GSK-3β和β-catenin的定位和阳性表达的变化,ELISA检测淀粉样蛋白(Aβ40/42)水平。结果Curcumin处理后,细胞内GSK-3β mRNA表达水平和蛋白水平都明显减弱;而GSK-3β的磷酸化形式GSK-3β-Ser9蛋白却明显增加,具有浓度-时间依赖性(P<0.05)。β-catenin和CyclinD1 mRNA表达水平和蛋白水平增加,呈浓度-时间依赖性(P<0.05)。免疫荧光染色结果不但证实了GSK-3β、β-catenin在细胞质内量的变化,而且还发现随着药物浓度的增加,β-catenin逐渐向细胞核内转移。Aβ40/42生成水平明显降低,呈浓度-时间依赖性(P<0.05)。结论GSK-3β是一个潜在性的AD治疗靶点,Curcumin能够通过抑制GSK-3β的活性来发挥防治AD的作用。Aim To explore the mechanisms of Curcumin which plays a role of protecting cells through regulating the activity of GSK-3β.Methods Plasmid APPswe and BACE1-mychis were transiently co-transfected into SHSY5Y cells.The cells were treated with Curcumin and then were collected to determine the mRNA and protein levels of GSK-3β,β-catenin and CyclinD1 by RT-PCR and Western blot. Immuno-fluoresecnt staining was used to detect the location and expression of β-catenin and GSK-3β, and ELISA was for the generation of Aβ.Results RT-PCR and Western blot results showed that in the cells, Curcumin inhibited the expression of GSK-3β mRNA and protein, and promoted the expression of GSK-3β-Ser9 protein （P0.05）.Meanwhile,Curcumin induced the expression of β-catenin and CyclinD1 mRNA and protein. All the data showed that all the changes also were in a concentration-and time-dependent manner （P0.05）. Immuno-fluoresecnt staining results not only confirmed the above changes, but also suggested that β-catenin had translocated into the nuclear gradually in a concentration-dependent manner.ELISA results showed that Curcumin significantly reduced the generation of Aβ40/42,which was in a concentration-and time-dependent manner （P0.05）.Conclusions GSK-3β is a potential target for treating with AD.Curcumin can prevent AD through inhibiting the activity of GSK-3β.

关 键 词：阿尔采末病 姜黄素 糖原合成激酶-3β Β-链蛋白 细胞周期蛋白D1 Β-淀粉样蛋白 

分 类 号：R282.71[医药卫生—中药学] R341[医药卫生—中医学]