针刺研究

2020, v.45(03) 194-201

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电针对阿尔茨海默病大鼠海马P35/P25-细胞周期素依赖性蛋白激酶5-Tau蛋白信号通路的影响
Effect of electroacupuncture on the P35/P25-cyclin-dependent kinase 5-Tau pathway in hippocampus of rats with Alzheimer’s disease

王依滢;高蒙;邱国平;徐进;伍艳君;许毅;王健蓉;盛华均;朱淑娟;
WANG Yi-ying;GAO Meng;QIU Guo-ping;XU Jin;WU Yan-jun;XU Yi;WANG Jian-rong;SHENG Hua-jun;ZHU Shu-juan;Department of Human Anatomy, Chongqing Medical University;College of Medicine, Chengdu University;Department of Neurology, The Fifth People's Hospital of Chengdu;

摘要(Abstract):

目的:通过观察电针对阿尔茨海默病(AD)大鼠P35/P25-细胞周期素依赖性蛋白激酶5(CDK5)-Tau蛋白信号通路的影响,探讨电针防治AD的机制。方法:SD大鼠随机分为对照组、假手术组、模型组和治疗组,每组12只。采用双侧海马注射Aβ_(25-35)复制AD大鼠模型。治疗组电针"百会"和双侧"肾俞"穴,每次15 min,每日1次,共10 d。用Morris水迷宫检测各组大鼠学习记忆能力,免疫组织化学法检测海马组织P35/P25、CDK5、Tau5蛋白分布及表达情况,Western blot法检测上述蛋白及Tau蛋白Ser199和Ser202位点的磷酸化表达水平。结果:在可视平台实验中,各组大鼠平均逃避潜伏期与搜索路径差异均无统计学意义(P>0.05)。在隐蔽平台实验中,与对照组比较,假手术组大鼠平均逃避潜伏期和搜索路径差异均无统计学意义(P>0.05);模型组平均逃避潜伏期和搜索路径较对照组、假手术组明显延长(P<0.05);与模型组比较,治疗组平均逃避潜伏期和搜索路径明显缩短(P<0.05)。在空间探索实验中,与对照组比较,假手术组穿越原平台次数差异无统计学意义(P>0.05);模型组穿越原平台的次数较对照组和假手术组显著减少(P<0.01,P<0.05);与模型组比较,治疗组穿越原平台次数显著增加(P<0.05)。与对照组及假手术组比较,模型组P35/P25、CDK5蛋白表达均明显升高(P<0.001);治疗组P35/P25、CDK5蛋白表达较模型组明显降低(P<0.001)。各组间Tau5蛋白表达差异无统计学意义(P>0.05)。Western blot法和免疫组织化学法的结果趋势一致。模型组大鼠海马区Tau蛋白Ser199和Ser202位点的磷酸化表达水平明显高于对照组和假手术组(P<0.01,P<0.05);治疗组Tau蛋白Ser199和Ser202位点的磷酸化表达水平较模型组明显降低(P<0.05,P<0.01)。结论:电针刺激可能通过影响大鼠海马区P35/P25-CDK5-Tau信号通路相关蛋白的表达变化,从而延缓AD的进展。
Objective To investigate the effect of electroacupuncture on the P35/P25-cyclin-dependent kinase 5(CDK5)-Tau pathway in rats with Alzheimer's disease(AD), as well as the mechanism of electroacupuncture in the prevention and treatment of AD. Methods Sprague-Dawley rats were randomly divided into control group, sham-operation group, model group, and electroacupuncture treatment group, with 12 rats in each group. A rat model of AD was established by injection of Aβ_(25-35) into the bilateral hippocampus. The rats in the electroacupuncture treatment group were given electroacupuncture at "Baihui"(GV20) and "Shenshu"(BL23) once a day, 15 min each time, for 10 days. Morris water maze was used to evaluate learning and memory abilities, immunohistochemistry was used to measure the distribution and expression of P35/P25, CDK5, and Tau5 in the hippocampus, and Western blot was used to measure the expression of the above mentioned proteins, phosphory-lated Tau(Ser199, Ser202). Results In the visual platform test, there were no significant differences in escape latency and search path between groups(P>0.05). In the hidden platform test, there were no significant differences in escape latency and search path between the control group and the sham-operation group(P>0.05); the model group had significantly longer escape latency and search path than the control group and the sham-operation group(P<0.05). Compared with the model group, the electroacupuncture treatment group had significantly shorter escape latency and search path(P<0.05). In the spatial exploration test, there was no significant difference in the number of platform crossings between the control group and the sham-operation group(P<0.05). The model group had a significantly lower number of platform crossings than the control group and the sham-operation group(P<0.01, P<0.05). The electroacupuncture treatment group had a significantly higher number of platform crossings than the model group(P<0.05). Compared with the control group and the sham-operation group, the model group had significant increases in the protein expression of P35/P25 and CDK5(P<0.001), and the electroacupuncture treatment group had significant reductions compared with the model group(P<0.001). There was no significant difference in the protein expression of Tau5 between groups(P>0.05). The model group had significantly higher protein expression of phosphorylated Tau(Ser199, Ser202) in the hippocampus than the control group and the sham-operation group(P<0.01, P<0.05). The electroacupuncture treatment group had significantly lower protein expression of phosphorylated Tau(Ser199,Ser202) than the model group(P<0.05, P<0.01). Conclusion Electroacupuncture may delay the progression of AD by affecting the expression of proteins involved in the P35/P25-CDK5-Tau pathway in the hippocampus of rats.

关键词(KeyWords): 电针;阿尔茨海默病;海马;P35/P25-CDK5-Tau蛋白信号通路
Electroacupuncture;Alzheimer's disease;Hippocampus;P35/P25-cyclin-dependent kinase 5-Tau pathway

Abstract:

Keywords:

基金项目(Foundation): 重庆市市科委项目(No.cstc2017jcyjAX0414、cstc2016jcyjA0229);; 重庆市教委项目(No.KJ1600213)

作者(Author): 王依滢;高蒙;邱国平;徐进;伍艳君;许毅;王健蓉;盛华均;朱淑娟;
WANG Yi-ying;GAO Meng;QIU Guo-ping;XU Jin;WU Yan-jun;XU Yi;WANG Jian-rong;SHENG Hua-jun;ZHU Shu-juan;Department of Human Anatomy, Chongqing Medical University;College of Medicine, Chengdu University;Department of Neurology, The Fifth People's Hospital of Chengdu;

Email:

DOI: 10.13702/j.1000-0607.190415

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