针刺研究

2018, v.43(04) 215-220

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“益肾调督”电针法对阿尔茨海默病小鼠大脑海马老年斑形成的影响
Kidney-reinforcing and Governor Vessel-regulating EA Intervention May Improve Learning-memory Possibly by Suppressing Formation of Senile Plaques in Hippocampus in APP/PS 1 Double Transgenic Alzheimer's Disease Mice

杨清华;郭玲;陈清;邬开会;伍艳君;贾岩;朱淑娟;唐成林;盛华均;
YANG Qing-hua;GUO Ling;CHEN Qing;WU Kai-hui;WU Yan-jun;JIA Yan;ZHU Shu-juan;TANG Cheng-lin;SHENG Hua-jun;Department of Teaching and Research of Anatomy,Institute of Neuroscience,Chongqing Medical University;C ollege of Traditional Chinese Medicine,Chongqing Medical University;

摘要(Abstract):

目的:观察"益肾调督"电针法对APP/PS 1双转基因阿尔茨海默病(AD)小鼠海马区老年斑(SP)及其相关蛋白表达的影响,探讨电针改善AD的机制。方法:APP/PS 1双转基因AD小鼠随机分为模型组、电针两疗程组和电针三疗程组,每组6只;6只雄性野生型小鼠为对照组。给予各电针组小鼠"百会"和双侧"肾俞"电针治疗,每天1次,7d为1个疗程,分别进行2个或3个疗程。Morris水迷宫实验观察各组小鼠记忆和空间探索能力,免疫组化法检测海马区SP的表达情况,Western blot法检测海马区淀粉样前体蛋白(APP)、β-分泌酶1(BACE 1)和胰岛素降解酶(IDE)的表达水平。结果:与对照组相比,模型组小鼠的逃避潜伏期和搜索路径均明显增加(P<0.01,P<0.05),穿越原平台的次数明显减少(P<0.01);与模型组相比,两治疗组小鼠的第5天逃避潜伏期和第4天、第5天搜索路径均明显缩短(P<0.01),穿越原平台的次数明显增加(P<0.01);与电针两疗程组相比,电针三疗程组的第5天逃避潜伏期和第4天、第5天搜索路径也明显缩短(P<0.05,P<0.01),穿越原平台的次数明显增加(P<0.05)。对照组小鼠海马区无SP阳性斑块;模型组SP数目较对照组明显增加(P<0.01);与模型组相比,两治疗组海马区的SP数目均明显减少(P<0.01);与电针两疗程组相比,电针三疗程组SP数目明显减少(P<0.01)。模型组小鼠海马区APP、BACE 1表达水平明显高于对照组(P<0.01),IDE表达水平明显低于对照组(P<0.01);两治疗组海马区APP、BACE 1表达水平明显低于模型组(P<0.01),IDE表达水平明显高于模型组(P<0.01);电针三疗程组海马区APP、BACE 1表达水平明显低于电针两疗程组(P<0.01,P<0.05),IDE表达水平明显高于电针两疗程组(P<0.05)。结论:"益肾调督"电针法可降低AD小鼠海马区APP、BACE 1表达,提高IDE的表达,从而减少海马区SP的沉积,改善其学习记忆和空间探索能力。
Objective To observe the effect of electroacupuncture(EA)intervention on learning-memory ability and the expression of senile plaques(SP),amyloid precursor protein(APP),β-secretase 1(BACE 1)and insulin degrading enzyme(IDE)in the hippocampus in APP/presenilin 1(PS 1)double transgenic Alzheimer's disease(AD)mice,so as to reveal its mechanisms underlying improvement of AD.Methods A total of 18 male APP/PS 1 double transgenic AD mice were randomly divided into model,EA-2-week and EA-3-week groups(n=6 in each).The control group was consisted of 6 male wild mice.EA(2 Hz,2 mA)was applied to"Baihui"(GV 20)and bilateral"Shenshu"(BL 23)for 15 min,once a day,with 7 days being a therapeutic course,2 or 3 courses altogether and with an one day's interval between every two courses.The spatial learningmemory ability was assessed using Morris water maze test during 5 days' training.The immunoactivity of SP in the hippocampus tissue was detected by immunohistochemistry,and the expression levels of APP,BACE 1 and IDE in the hippocampus were analyzed by Western blot.Results Following modeling,the escape latency and path length of hidden platform tests were significantly increased(P<0.01,P<0.05),and the platform crossing time of spatial probing test significantly decreased(P<0.01)in the model group compared with the control group.After EA intervention,the escape latency on the 5 th day of training,and the path length on the 4 th and 5 th day of training in both EA-2-week and EA-3-week groups were significantly shorter relevant to the model group(P<0.01),and those of the EA-3-week group were considerably shorter than those of the EA-2-week group in the escape latency and path length(P<0.05,P<0.01).The platform crossing times of spatial probing test were significanthy increased in both EA-2-week and EA-3-week groups in comparison with the model group(P<0.01),and that of the EA-3-week group was considerably increased compared with the EA-2-week group(P<0.05).Immunohistochemical staining showed that the number of SP in the hippocampus was markedly increased in the model group compared with the control group(P<0.01),and was markedly reduced in both EA-2-week and EA-3-week groups(P<0.01),and that of the EA-3-week group was significantly decreased compared with the EA-2-week group(P<0.01).The expression levels of hippocampal APP and BACE 1 proteins were significantly higher in the model group than in the control group(P<0.01),and that of hippocampal IDE was markedly lower in the model group than in the control group(P<0.01).After EA,the increased expression levels of APP and BACE 1 proteins and the decreased expression level of IDE in the EA-2-week and EA-3-week groups were significantly inhibited(P<0.01).The effects of EA-3-week were significantly stronger than those of EA-2-week in down-regulating the expression of APP and BACE 1 proteins and up-regulating the expression of IDE(P<0.01,P<0.05).Conclusion EA stimulation of GV 20 and BL 23 can improve the learning-memory ability in APP/PS 1 double transgenic AD mice,which may be related to its effects in down-regulating the expression of SP,APP and BACE 1 proteins and up-regulating the expression of IDE protein in the hippocampus.

关键词(KeyWords): APP/PS 1双转基因阿尔茨海默病小鼠;电针;学习记忆能力;老年斑;淀粉样前体蛋白;β-分泌酶;胰岛素降解酶;海马
APP/presenilin 1(PS 1)double transgenic Alzheimer's disease mice;Electroacupuncture;Learning-memory ability;Senile plaques;Amyloid precusor protein;β-secretase;Insulin degrading enzyme;Hippocampus

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(No.81273870);; 重庆市科委基础与前沿项目(No.cstc 2014jcyj A 10028);; 重庆市渝中区科技计划项目(No.20150122);; 重庆医科大学基础医学院“英耀计划”项目(No.JCYY 201610)

作者(Author): 杨清华;郭玲;陈清;邬开会;伍艳君;贾岩;朱淑娟;唐成林;盛华均;
YANG Qing-hua;GUO Ling;CHEN Qing;WU Kai-hui;WU Yan-jun;JIA Yan;ZHU Shu-juan;TANG Cheng-lin;SHENG Hua-jun;Department of Teaching and Research of Anatomy,Institute of Neuroscience,Chongqing Medical University;C ollege of Traditional Chinese Medicine,Chongqing Medical University;

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DOI: 10.13702/j.1000-0607.170460

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