基于内体-溶酶体系统探讨电针“百会”“涌泉”改善APP/PS1双转基因小鼠学习记忆能力的作用机制  

Electroacupuncture at “Baihui”(GV20) and “Yongquan”(KI1) improves learning-memory ability of APP/PS1 double transgenic mice by regulating endosomal-lysosomal system

作　　者：栗晨璐 杨晓坤 高誉珊 郭孟玮[1] 谭韵湘 张洋 陈浩天 薛卫国[1] LI Chen-lu;YANG Xiao-kun;GAO Yu-shan;GUO Meng-wei;TAN Yun-xiang;ZHANG Yang;CHEN Hao-tian;XUE Wei-guo(School of Acupuncture-Moxibustion and Tuina,Beijing University of Chinese Medicine,Beijing 100029,China;School of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学针灸推拿学院,北京100029 [2]北京中医药大学中医学院,北京100029

出　　处：《针刺研究》2023年第8期791-798,共8页Acupuncture Research

基　　金：北京市自然科学基金项目(No.7202117)。

摘　　要：目的:观察电针“百会”“涌泉”对APP/PS1双转基因小鼠学习记忆能力、海马区β淀粉样蛋白沉积及相关蛋白表达的影响,从内体-溶酶体系统角度探讨电针改善阿尔茨海默病(AD)的作用机制。方法:将6月龄雄性APP/PS1转基因小鼠随机分为模型组和电针组,每组10只;另以10只6月龄雄性C57BL/6小鼠作为正常组。电针组针刺“百会”“涌泉”,双侧“涌泉”接电针,15 min/次;模型组和正常组在相同时间以相同方法束缚15 min/次。以上干预均隔日1次,共6周。以Morris水迷宫实验检测各组小鼠学习记忆能力,免疫组织化学法检测小鼠海马区老年斑(SP)的阳性表达,透射电镜观察小鼠海马神经元内体、溶酶体超微结构,Western blot法检测小鼠海马Ras相关蛋白5(Rab5)、Ras相关蛋白7(Rab7)、组织蛋白酶D(CTSD)等内体-溶酶体系统相关蛋白的表达水平。结果:与正常组比较,模型组小鼠逃避潜伏期延长(P<0.05,P<0.01),穿越原平台次数及原平台象限游泳路程均减少(P<0.05),海马区SP阳性表达增加(P<0.01),海马神经元胞体中内体、溶酶体堆积,海马组织Rab5、Rab7、CTSD蛋白表达水平均升高(P<0.01)。与模型组比较,电针组小鼠逃避潜伏期缩短(P<0.05),穿越原平台次数及原平台象限游泳路程均增加(P<0.05),海马区SP阳性表达减少(P<0.01),海马神经元胞体中内体、溶酶体堆积减少,海马组织Rab5、Rab7、CTSD蛋白表达水平均降低(P<0.01)。结论:电针“百会”“涌泉”可改善AD小鼠学习记忆能力,减少海马区β淀粉样蛋白沉积,其机制可能与电针调控内体-溶酶体系统功能有关。Objective To explore the mechanism of electroacupuncture(EA) in improving learning-memory ability in Alzheimer's disease(AD) mice from the perspective of endosomal-lysosomal system.Methods Male APP/PS1 transgenic mice were randomly divided into model group and EA group(n=10 in each group) and 10 male C57BL/6wild mice were taken as the normal group.EA(1 Hz/50 Hz,1 mA) was applied at bilateral “Yongquan”(KI1) and acupuncture was applied at “Baihui”(GV20) for 15 min.The mice of the model and normal groups were subjected to restriction with the same method as those of the EA group for 15 min.The treatment was conducted once every other day for 6 weeks.The spatial learning-memory ability(shown by escape latency of place navigation test and the time of crossing the target platform and total swimming distance in the target quadrant in 1 min of spatial probe test) was detected by Morris water maze test.The immunoactivity of senile plaques(SP) in the hippocampus tissue was detected by immunohistochemistry.The ultrastructural characters of hippocampal neurons were observed by transmission electron microscope,and the expression levels of Ras-related protein 5(Rab5),Ras-related protein 7(Rab7) and cathepsin D(CTSD) in the hippocampus were detected by Western blot,separately.Results Compared with the normal group,the escape latency,SP immunoactivity,and protein expression levels of Rab5,Rab7 and CTSD were significantly increased(P<0.05,P<0.01),while the number of crossing the original platform and the total swimming distance in the platform quadrant were considerably reduced(P<0.05) in the model group.In contrast to the model group,the EA group had a marked decrease in the escape latency,SP immunoactivity,and protein expression levels of Rab5,Rab7 and CTSD(P<0.05,P<0.01),and a striking increase in the number of crossing the original platform and the swimming distance in the platform quadrant(P<0.05).Results of transmission electron microscope showed an accumulation of endosome,lysosome,and endolysosomes in the hippoca

关 键 词：阿尔茨海默病 电针 内体-溶酶体系统 学习记忆能力 老年斑 海马 

分 类 号：R245.97[医药卫生—针灸推拿学]