糖原合酶激酶-3导致阿尔茨海默病模型大鼠认知障碍的机制研究  

The mechanism study on the effect of glycogen synthase kinase-3 on cognitive function in rats model with Alzheimer’s disease

作　　者：王玥[1] 胡文立[1] 李淑娟[1] 杨秋实[2] 赵丽[2] 

机构地区：[1]首都医科大学附属北京朝阳医院神经内科,北京市100020 [2]首都医科大学基础医学院神经生物学系,北京市100069

出　　处：《老年医学与保健》2015年第6期350-353,358,共5页Geriatrics & Health Care

基　　金：国家自然科学基金(81301101、81400948);北京市自然科学基金(7144188);北京市教育委员会科技计划面上项目(KM201410025004);首都医科大学基础临床合作研究基金(13JL25);首都医科大学省部级重点实验室开放研究课题(GJZDSS004)

摘　　要：目的探讨糖原合酶激酶-3（GSK-3）对老年阿尔茨海默病（AD）模型大鼠认知功能及海马乙酰胆碱（ACh）水平的影响，从而找寻AD记忆障碍的机制。方法30只Wistar大鼠随机分为3组，每组10只大鼠：侧脑室注射人工脑脊液（aCSF）组（Vehicle组）、侧脑室注射Wortmannin组（Wortmannin组）和侧脑室联合注射Wortmannin和LiCl组（Wortmannin＋LiCl组）。每组10只。利用Morris水迷宫检测大鼠空『白J记忆功能；侧脑室注射P13K抑制剂Wortmannin后，利用放射性同位素方法检测GSK-3活性，利用免疫印迹检测GSK-3及tau蛋白磷酸化水平；利用微量渗析及高压液相色谱方法检测ACh水平。结果侧脑室注射Wortmannin后大鼠海马GSK-3活性显著升高，GSK-3D在Ser9位点及tau蛋白在Ser396、Thr231位点磷酸化水平显著改变，说明激活的GSK-3B可以磷酸化tau蛋白并诱导AD样病理改变。侧脑室注射Wortmannin后大鼠表现出明显的空间记忆障碍，同时海马ACh水平显著降低。然而，如同时注射GSK-3抑制剂LiCI后，ACh则恢复到正常水平。结论侧脑室注射Wortmarmin可以激活海马GSK-3活性，激活的GSK-3可以通过下调ACh诱导老年AD大鼠空间记忆障碍。Objective To investigate the effect of glycogen synthase kinase-3 （GSK-3） on cognitive function and acetylcholine （ACh） level in hippocampus in rats model in order to find the memory deficit mechanism of Alzheimer＇s disease （AD）. Methods 30 Wistar rats were divided into 3 groups （every group had I0 rats）： one group was injected into lateral ventricle with artificial cerebrospinal fluid （Vehicle group）, another group with Wortmannin （Wortmannin group）, the third group with Wortmannin and LiC1 （Wortmannin ＋ LiCI group）. Morris water maze was used to detect spatial memory in rats. After ventricular injection of Wortmannin, GSK-3 activity was measured by radioisotope 32p method, phosphorylation levels of GSK-3 and tau were measured by Western blot, and the ACh level was detected by microdialysis and high-performance liquid chromatography （HPLC）. Results GSK-3 was activated in rat hippocampus after ventricular injection of Wortmannin, a PI3K inhibitor. Wortmannin increased remarkably the phosphorylation level of GSK-3 at Ser9 epitope, Tau at Ser396 and Thr231 epitope, which indicated that GSK-3 phosphorylated substrates and induced AD-like pat- hological alterations successfully. Spatial memory deficit in rats was induced by ventricular injection of Wortmannin. At the same time, the ACh level in hippocampus decreased significantly, which returned to normal level when LiCl was added. Conclusion GSK-3 was activated after ventricular injection of Wortmaunin. The activated GSK-3 induced spatial memory deficit in rats by down regulation of ACh level.

关 键 词：阿尔茨海默病 认知障碍 糖原合酶激酶-3 乙酰胆碱 TAU蛋白 海马 

分 类 号：R749.16[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]