摘要
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.
作者
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;College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing 400016, China)
出处
《针刺研究》
CAS
CSCD
北大核心
2018年第4期215-220,共6页
Acupuncture Research
基金
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)
关键词
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
I5-secretase
Insulin degrading enzyme
Hippocampus
