电针对阿尔茨海默病不同病理阶段自噬状态影响差异性研究  

Study of the effect differences of electroacupuncture on autophagy state in different pathological stages of Alzheimer's disease

作　　者：杨晓坤 高誉珊[1] 栗晨璐 陈浩天 郭孟玮[1] 谭韵湘 张洋[1] 薛卫国[1] YANG Xiaokun;GAO Yushan;LI Chenlu;CHEN Haotian;GUO Mengwei;TAN Yunxiang;ZHANG Yang;XUE Weiguo(Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学,北京100029

出　　处：《上海针灸杂志》2023年第8期854-863,共10页Shanghai Journal of Acupuncture and Moxibustion

基　　金：北京市自然科学基金资助项目面上项目(7202117)。

摘　　要：目的 观察电针对APP/PS1双转基因小鼠老年斑出现前后不同病理阶段自噬状态影响的差异性,从自噬角度探讨电针干预阿尔茨海默病(Alzheimer’s disease,AD)的作用机制。方法 4月龄和8月龄APP/PS1双转基因雄性小鼠各20只,随机分为模型组(4月龄和8月龄模型组)和电针组(4月龄和8月龄电针组),每组10只;以同月龄同性别C57BL/6野生型小鼠为正常组(4月龄和8月龄正常组)各10只。电针组取双侧涌泉穴直刺后接电针仪,百会穴平刺后单向滞针再留针,共20 min。干预隔日1次,共6周。采用Morris水迷宫实验评价小鼠空间学习记忆能力;通过免疫组化技术观察小鼠海马淀粉样斑块沉积情况;用免疫荧光技术观察小鼠海马LC3和β-淀粉样蛋白(β-amyloid,Aβ)表达情况;用酶联免疫吸附法检测小鼠海马Aβ表达水平;用Western blot法检测小鼠海马自噬相关蛋白LC3和p62相对表达量。结果 Morris水迷宫结果显示,6月龄和10月龄模型组小鼠平均逃避潜伏期均较同月龄正常组增加(P<0.01),平台象限停留时间均减短(P<0.01);与6月龄和10月龄模型组相比,同月龄电针组小鼠平均逃避潜伏期均减少(P<0.05),平台象限停留时间均延长(P<0.01)。免疫组化结果显示,6月龄模型组海马区域可见少量老年斑,10月龄模型组海马区域可见大量老年斑。免疫荧光技术结果显示,LC3和Aβ阳性表达主要位于海马神经元核周,LC3阳性还表达在细胞突起中。酶联免疫吸附法检测结果显示,同月龄模型组海马Aβ水平均高于正常组(P<0.01);同月龄电针组海马Aβ水平均低于同月龄模型组(P<0.01);6月龄模型组海马Aβ水平明显低于10月龄模型组(P<0.01)。Western blot法检测结果显示,与同月龄正常组比较,6月龄模型组LC3Ⅱ/Ⅰ比值和p62表达均降低(P<0.01);与同月龄模型组比较,电针组LC3Ⅱ/Ⅰ比值增加(P<0.01),p62差异无统计学意义(P>0.05)。与同月龄正常组比较,10月龄模型组LObjective To observe the effect differences of electroacupuncture(EA)on the autophagy state in APP/PS1 double transgenic mice in different pathological stages,before and after the occurrence of senile plaques,and to explore the mechanism of EA treatment of Alzheimer’s disease(AD)from the perspective of autophagy.Method Four-month-old and 8-month-old APP/PS1 double transgenic male mice,20 in each age group,were randomized into model groups(4-month-old model group and 8-month-old model group)and EA groups(4-month-old EA group and 8-month-old EA group),with 10 mice in each group.Wild-type C57BL/6 mice of the same ages and gender were taken as control groups(4-month-old and 8-month-old normal groups),with 10 mice in each group.The EA groups received EA at bilateral Yongquan(KI1)and Baihui(GV20)was horizontally punctured with the needle twirled till stuck and then retained for 20 min.The intervention was performed once every other day for 6 weeks.Morris water maze test was used to evaluate the spatial learning and memory abilities of the mice;immunohistochemistry was taken to observe the deposition of amyloid plaques in the hippocampus of mice;the immunofluorescence technique was employed to detect the expression of LC3 andβ-amyloid(Aβ)in the hippocampus of mice;the enzyme-linked immunosorbent assay(ELISA)was used to determine the expression level of Aβin the hippocampus of mice;Western blot(WB)was used to detect the relative expression levels of autophagy-related proteins LC3 and p62 in the hippocampus.Result Morris water maze results showed that compared with the normal groups,the 6-month-old and 10-month-old model groups had a longer average escape latency(P<0.01)and shorter platform dwelling time(P<0.01);compared with the 6-month-old and 10-month-old model groups,mice in the same age EA groups had a reduced average escape latency(P<0.05)and extended platform dwelling time(P<0.01).The immunohistochemistry results showed a small amount of senile plaques in the hippocampus of 6-month-old model mice and a large a

关 键 词：针刺疗法 电针 阿尔茨海默病 APP/PS1双转基因小鼠 自噬 

分 类 号：R2-03[医药卫生—中医学]