还脑益聪方组分对β淀粉样前体蛋白转基因痴呆小鼠行为学及胆碱能系统的早期干预作用  

Effects of early intervention with Huannao Yicong formula effective components on behavior and cholinergic system of β-amyloid precursor protein transgenic mice

作　　者：蔡琳琳[1] 李浩[1] 刘剑刚[1] 刘龙涛[1] 官杰 刘明芳[1] 胡佳[3] 魏芸[3] 

机构地区：[1]中国中医科学院西苑医院老年病中心,北京100091 [2]山东省青岛市海慈医疗集团内分泌科,山东青岛266033 [3]北京化工大学化工资源有效利用国家重点实验室,北京100029

出　　处：《中西医结合学报》2011年第3期292-298,共7页Journal of Chinese Integrative Medicine

基　　金：国家自然科学基金资助项目(No.30873338);国家科技重大专项课题资助项目(No.2009ZX09103-391)

摘　　要：目的:观察还脑益聪方复方组分早期干预对阿尔茨海默病(Alzhei mer disease,AD)模型β淀粉样前体蛋白(β-amyloid precursor protein,APP)转基因小鼠行为学及脑胆碱能系统的影响。方法:将60只3月龄APP695V717I转基因小鼠随机分为4组:模型组、大剂量还脑益聪方组[2.80g/(kg.d)]、小剂量还脑益聪方组[1.40g/(kg.d)]及盐酸多奈哌齐阳性对照组[0.65mg/(kg.d)],每组15只。15只遗传背景相同的非转基因C57BL/6J小鼠作为正常对照组。所试药物稀释至相同体积灌胃给药,正常组和模型组给以等体积蒸馏水灌胃,连续灌胃6个月。采用Morris水迷宫实验和避暗实验进行小鼠行为学测试,采用酶联免疫吸附测定法分别检测小鼠大脑皮层和海马组织的乙酰胆碱(acetylcholine,ACh)、乙酰胆碱酯酶(acetylcholinesterase,AChE)含量及胆碱乙酰转移酶(choline acetyltransferase,ChAT)的活性,采用考马斯亮蓝法测定脑组织中蛋白的含量。结果:与正常对照组比较,模型组小鼠学习记忆成绩均显著下降(P<0.05,P<0.01),海马和皮层组织的ACh含量和ChAT活性显著降低,AChE含量显著增加(P<0.01,P<0.05)。与模型组比较,盐酸多奈哌齐组、还脑益聪方组小鼠的学习记忆能力均有明显提高(P<0.05);盐酸多奈哌齐组、还脑益聪方大剂量组小鼠海马ACh含量明显提高(P<0.05),盐酸多奈哌齐组、还脑益聪方组小鼠海马AChE含量显著降低,ChAT活性显著升高(P<0.01,P<0.05);盐酸多奈哌齐组和还脑益聪方两组小鼠皮层ACh含量均明显提高(P<0.05),AChE含量有不同程度的降低,还脑益聪方两组小鼠皮层ChAT活性均明显提高(P<0.05)。结论:还脑益聪方能明显改善APP转基因小鼠的学习记忆能力,其作用机制可能与调控小鼠海马与皮层胆碱能系统相关酶的含量有关。Objective：To observe the effects of early intervention with effective components from a Chinese herbal formula（Huannao Yicong formula,HNYCF）on behavior and related indicators of cholinergic system in β-amyloid precursor protein（APP）transgenic mice.Methods：Sixty 3-month-old APP695V717I transgenic mice were randomly divided into model group,high-dose HNYCF group（2.80 g/（kg·d））,low-dose HNYCF group（1.40 g/（kg·d））and donepezil group（0.65 mg/（kg·d））,with 15 mice in each group.Fifteen non-transgenic mice of the same genetic background were used as normal group.The model group and normal group were fed with equal volume of distilled water by gavage.After 6-month continuous medication,the Morris water maze and the passive avoidance test were used to detect the visual spatial learning and memory ability of each mouse.Then the mice were decapitated and their cerebral cortex and hippocampus were isolated to homogenate by sonication.Contents of acetylcholine（ACh）and acetylcholinesterase（AChE）and choline acetyltransferase（ChAT）activity in the homogenate were determined by enzyme-linked immunosorbent assay,and protein contents of cerebral cortex and hippocampus were measured by Coomassie brilliant blue method.Results：Compared with the model group,high-and low-dose HNYCF and donepezil hydrochloride all improved spatial learning of APP mice in the Morris water maze.The ratio of swimming distance in the central area in the high-dose HNYCF group was longer than that in the model group（P0.05）.In the passive avoidance test,high-and low-dose HNYCF and donepezil hydrochloride improved memory function of APP mice by improving the escape latency and reducing the number of errors（P0.05,P0.01）.High-and low-dose HNYCF and donepezil hydrochloride reduced the content of AChE,increased the activity of ChAT（P0.01,P0.05）and improved the content of ACh in hippocampus（P0.05）;high-and low-dose HNYCF and donepezil hydrochloride increased the content of ACh in cortex（P0.05）.Do

关 键 词：阿尔茨海默病 中草药 淀粉样Β蛋白前体 动物 基因修饰 神经行为学表现 乙酰胆碱 乙酰胆碱酯酶 胆碱O-乙酰转移酶 小鼠 

分 类 号：R285.5[医药卫生—中药学]