清心开窍方对APP/PS1双转基因小鼠学习记忆及海马CA1区Akt、GSK3α、βAPP、Aβ表达的影响  

Effect of Qingxin Kaiqiao Decoction on Learning-memory Ability and Expressions of Akt, GSK3α, βAPPand Aβ on Hippocampal CA1 Region of APP/PS1 Double Transgenic Mice

作　　者：李燕[1] 王天琪 林坚炜 高诗雨 杨文育 胡海燕[1,2] LI Yan;WANG Tianqi;LIN Jianwei;GAO Shiyu;YANG Wenyu;HU Haiyan(The Second Clinical College of Wenzhou Medical University, Wenzhou 325003 , Zhejiang,China;Department of Traditional Chinese Medicine, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325003 , Zhejiang,China)

机构地区：[1]温州医科大学第二临床学院,浙江温州325003 [2]温州医科大学附属第二医院中医科,浙江温州325003

出　　处：《中华中医药学刊》2019年第5期1128-1132,I0014-I0019,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81573896);浙江省自然科学基金项目(LY15H270016)

摘　　要：目的:探讨清心开窍方对APP/PS1双转基因小鼠学习记忆能力、海马CA1区神经元形态学的改变,以及对Akt、GSK3α、βAPP、Aβ蛋白表达的影响。方法:选取50只3月龄雄性APP/PS1双转基因小鼠,随机分为模型组、多奈哌齐组(1.67 mg·kg^(-1))、清心开窍方高、中、低剂量组(每日给药剂量分别为19、9.5、4.75 g·kg^(-1)),另取10只同背景同性别同月龄的C57BL/6J小鼠作为对照组,对照组、模型组给予等体积生理盐水灌胃,1次/d,连续灌胃12周。采用Morris水迷宫检测小鼠空间学习记忆能力,透射电镜观察小鼠海马CA1区神经元超微结构,尼氏染色法观察小鼠海马CA1区神经元损伤情况,免疫组织化学染色法检测海马CA1区Akt、GSK3α、βAPP、Aβ蛋白的表达情况。结果:与对照组比较,模型组APP/PS1双转基因小鼠逃避潜伏期延长(P<0.01),穿台次数减少,其海马CA1区神经元超微结构出现严重损伤,锥体细胞数量减少,排列紊乱,尼氏体着色变浅,Akt蛋白表达减少(P<0.01),GSK3α、βAPP、Aβ表达增加(P<0.01);与模型组比较,清心开窍方干预组逃避潜伏期均明显缩短(P<0.01),穿台次数增多,同时比较小鼠在原平台停留的时间和路程百分比均明显增加(P<0.05;P<0.01),其神经元结构较完整,核膜清晰,胞浆内核糖体丰富,线粒体数量增多,且嵴结构完整,锥体细胞数量增多,排列紧密,尼氏体着色较深,同时Akt蛋白表达增加(P<0.01;P<0.05),GSK3α、βAPP、Aβ表达减少(P<0.01;P<0.05)。结论:清心开窍方能够改善APP/PS1双转基因小鼠的学习记忆能力,其机制可能与Akt/GSK3α介导的信号通路相关,它能够激活Akt,抑制GSK3α,降低βAPP、Aβ的生成,减少神经元损伤。Objective: This study was aimed to investigate the changes of spatial cognitive function with Qingxin Kaiqiao Decoction(QKD) in APP/PS1 transgenic mice, the pathological changes of neurons and the expressions of Akt,GSK3α,βAPP and Aβ in hippocampus CA1 region and then explore the possible mechanism of QKD’s anti-AD function. Methods: Totally 50 three-month-old male APP/PS1 transgenic mice were used to establish the AD model. They were randomly divided into the model group, the Donepezil group, the QKD high dose group(19 g·kg^-1), the QKD middle dose group(9.5 g·kg-1) and the QKD low dose group(4.75 g·kg^-1). Besides, 10 three-month-old male C57 BL/6 J mice were selected as the normal control group. The control group and the model group were given equal volume of saline. All mice were gavaged for 12 consecutive weeks. At the end of medication, all mice were tested for their capabilities of spatial navigation and memorization by Morris water maze. The ultrastructure of hippocampus neuronal cells was observed by transmission electron microscopy(TEM)and its nissl body was tested by Nissl staining. Immunohistochemical staining was employed to confirm the expressions of Akt, GSK3α,βAPP and Aβ in hippocampus CA1 region. Results: Compared with the normal group, the escape latency of the APP/PS1 double transgenic mice in the model group were prolonged(P<0.01), while the number of crossing the platform was decreased. In the hippocampal CA1 area, the ultrastructure of neurons were severely damagedand the number of pyramidal cells were decreased while it arranged disorder and stained lightly. The protein expressions of Akt were decreased(P<0.01). GSK3α,βAPPand Aβ were increased(P<0.01). Compared with the model group, the escape latency of QKD groups was significantly shortened(P<0.01), the number of crossing the platform was increasedand the percentage of swimming distancy and swimming time in the target quadrant were significantly increased(P<0.05,P<0.01). Compared with the model group, the neural structure of

关 键 词：阿尔茨海默病 清心开窍方 丝氨酸苏氨酸蛋白激酶(Akt) 糖原合成酶激酶-3α(GSK3α) β淀粉样前体蛋白(βAPP) β-淀粉样蛋白(Aβ) 

分 类 号：R285.5[医药卫生—中药学]