电针对阿尔茨海默病大鼠学习记忆能力及前额叶P35/P25-周期蛋白依赖性激酶5-Tau蛋白磷酸化通路的影响

doi:10.3969/j.issn.1006-9771.2020.00.006

《中国康复理论与实践》 ›› 2020, Vol. 26 ›› Issue (6): 654-661.doi: 10.3969/j.issn.1006-9771.2020.00.006

电针对阿尔茨海默病大鼠学习记忆能力及前额叶P35/P25-周期蛋白依赖性激酶5-Tau蛋白磷酸化通路的影响

王依滢^1a,唐成林^1b,邱国平^1a,徐进^1a,隆令^1a,许毅²,王健蓉³,甘胜伟^1a,盛华均^1a,朱淑娟^1a()

1.重庆医科大学, a.人体解剖教研室; b.中医药学院, 重庆市 400016
2.成都大学医学院, 四川成都市 610106
3.成都市第五人民医院神经内科, 四川成都市 611130

收稿日期:2019-12-26 修回日期:2020-02-03 出版日期:2020-06-25 发布日期:2020-06-29
通讯作者: 朱淑娟 E-mail:shujuanzhu@cqmu.edu.cn
作者简介:王依滢(1995-),女,汉族,重庆市人,硕士研究生,主要研究方向：神经退行性疾病的防治研究。
基金资助:
1.国家自然科学基金面上项目(81273870);2.重庆市市科委项目(cstc2017jcyjA0414);重庆市市科委项目(cstc2016jcyjA0229);重庆市市科委项目(cstc2016jcyjA0073);3.重庆市教委项目(KJ1600213)

Effects of Electroacupuncture on Learning-memory and P35/P25-Cyclin-dependent Kinase 5-Tau Phosphorylation Signal Pathway in Prefrontal Cortex of Rats with Alzheimer's Disease

WANG Yi-ying^1a,TANG Cheng-lin^1b,QIU Guo-ping^1a,XU Jin^1a,LONG Ling^1a,XU Yi²,WANG Jian-rong³,GAN Sheng-wei^1a,SHENG Hua-jun^1a,ZHU Shu-juan^1a()

1.a. The Department of Human Anatomy; b. College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing 400016, China
2. College of Medicine, Chengdu University, Chengdu, Sichuan 610106, China
3. Department of Neurology, the Fifth People’s Hospital of Chengdu, Chengdu, Sichuan 611130, China

Received:2019-12-26 Revised:2020-02-03 Published:2020-06-25 Online:2020-06-29
Contact: ZHU Shu-juan E-mail:shujuanzhu@cqmu.edu.cn
Supported by:
National Natural Science Foundation of China(81273870);Science Foundation Project of CQCSTC(cstc2017jcyjA0414);Science Foundation Project of CQCSTC(cstc2016jcyjA0229);Science Foundation Project of CQCSTC(cstc2016jcyjA0073);Scientific and Technological Research Program of Chongqing Municipal Education Commission(KJ1600213)

摘要/Abstract

摘要：

目的观察电针百会、肾俞两穴对阿尔茨海默病(AD)模型大鼠学习记忆能力及前额叶皮质(PFC) P35/P25-周期蛋白依赖性激酶5 (CDK5)-Tau蛋白磷酸化信号通路的影响,以探讨电针治疗AD的相关机制。方法雄性成年Sprague-Dawley大鼠随机分为正常对照组、假手术组、模型组和电针组,每组6只。后两组双侧海马注射Aβ_25-35,假手术组注射等量生理盐水。造模后次日,电针组电针百会、肾俞,留针15 min,每天1次,共10 d。治疗后,行Morris水迷宫实验,免疫组织化学染色和Western blotting检测各组PFC P35/P25-CDK5-Tau蛋白磷酸化相关蛋白的表达情况。结果与正常对照组和假手术组比较,模型组的逃避潜伏期和搜索路径均增加(P < 0.05),穿越平台次数减少( P < 0.05);与模型组比较,电针组逃避潜伏期和搜索路径均缩短( P < 0.05),穿越平台次数增加( P < 0.05)。模型组P35/P25和CDK5阳性表达明显高于正常对照组和假手术组( P < 0.01),电针组显著低于模型组( P < 0.001)。模型组P35/P25、CDK5、Tau[pS199]、Tau[pS202]相对表达量高于正常对照组和假手术组( P < 0.05),电针组上述蛋白的表达低于模型组( P < 0.05)。结论电针刺激能有效改善AD大鼠的学习记忆和空间探索能力,可能通过影响大鼠PFC的P35/P25-CDK5-Tau蛋白磷酸化信号通路,延缓AD的发生与发展。

关键词: 阿尔茨海默病, 电针, 前额叶皮质, P35/P25-周期蛋白依赖性激酶5-Tau蛋白磷酸化, 大鼠

Abstract:

Objective To observe the effect of electroacupuncture at Baihui (DU20) and Shenshu (BL23) acupoints on learning-memory ability and expression of the relative protein of P35/P25-cyclin-dependent kinase 5 (CDK5)-Tau phosphorylation signaling pathway in the prefrontal cortex (PFC) in rats with Alzheimer's disease (AD), so as to reveal its potential mechanisms in treating AD. Methods Male adult Sprague-Dawley rats were randomly divided into normal control group, sham group, model group and treatment group with six rats in each group. The AD model was constructed by bilateral hippocampal injection of Aβ_25-35 in latter two groups. Equal amount of normal saline was injected into the sham group. The treatment group was acupunctured at Baihui and Shenshu once a day for ten days. All the rats were tested with Morris Water Maze. Immunohistochemistry staining and Western blotting were used to detect the related protein of P35/P25-CDK5-Tau protein phosphorylation in the PFC.Results Compared with the normal control group and the sham group, the escape latency and escape length increased (P< 0.05) and the times crossing the platform reduced (P< 0.05) in the model group; compared with the model group, the escape latency and escape length reduced (P< 0.05), and the times crossing the platform increased (P< 0.05) in the treatment group. The optical density of P35/P25 and CDK5 were significantly higher in the model group than in the normal control group and the sham group (P< 0.01), and they were lower in the treatment group than in the model group (P< 0.001). The relative expression of P35/P25, CDK5, Tau[pS199] and Tau[pS202] were higher in the model group than in the normal control group and the sham group (P< 0.05), and the expression of the above proteins was lower in the treatment group than in the model group (P< 0.05).Conclusion Electroacupuncture could improve the learning-memory and spatial exploration ability, which associate with inhabiting the P35/P25-CDK5-Tau protein phosphorylation signaling pathway in the PFC to delay the development of AD.

Key words: Alzheimer's disease, electroacupuncture, prefrontal cortex, P35/P25-cyclin-dependent kinase 5-Tau phosphorylation, rats

中图分类号:

R749.1

王依滢,唐成林,邱国平,徐进,隆令,许毅,王健蓉,甘胜伟,盛华均,朱淑娟. 电针对阿尔茨海默病大鼠学习记忆能力及前额叶P35/P25-周期蛋白依赖性激酶5-Tau蛋白磷酸化通路的影响[J]. 《中国康复理论与实践》, 2020, 26(6): 654-661.

WANG Yi-ying,TANG Cheng-lin,QIU Guo-ping,XU Jin,LONG Ling,XU Yi,WANG Jian-rong,GAN Sheng-wei,SHENG Hua-jun,ZHU Shu-juan. Effects of Electroacupuncture on Learning-memory and P35/P25-Cyclin-dependent Kinase 5-Tau Phosphorylation Signal Pathway in Prefrontal Cortex of Rats with Alzheimer's Disease[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2020, 26(6): 654-661.

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组别	n	逃避潜伏期(s)	搜索路径(cm)
正常对照组	6	31.38±4.086	783.3±110.40
假手术组	6	37.30±2.591	815.3±67.10
模型组	6	36.78±4.032	890.7±101.20
电针组	6	33.14±3.898	719.4±66.82
F值		0.597	0.647
P值		0.624	0.593

组别	n	第3天	第4天	第5天	第6天
正常对照组	6	19.29±1.652	17.59±1.064	12.98±2.355	9.34±0.980
假手术组	6	23.14±2.211	18.22±2.162	13.24±2.199	11.64±2.117
模型组	6	35.71±2.896^a,b	33.80±2.056^a,b	30.43±3.222^a,b	24.79±3.192^a,b
电针组	6	19.67±3.316^c	19.17±2.320^c	13.49±1.505^c	12.64±2.991^c
F值	6	8.787	15.36	12.85	13.64
P值	6	0.001	< 0.001	< 0.001	< 0.001

组别	n	第3天	第4天	第5天	第6天
正常对照组	6	359.9±57.27	296.2±20.04	218.4±33.42	137.2±15.05
假手术组	6	392.0±38.6	335.5±30.55	219.8±30.86	202.3±32.48
模型组	6	683.3±106.2^a,b	587.6±39.98^a,b	481.4±62.57^a,b	370.8±47.82^a,b
电针组	6	353.9±65.62^c	304.3±61.18^c	248.6±27.25^c	160.2±25.24^c
F值	6	4.919	11.550	9.591	10.610
P值	6	0.013	< 0.001	< 0.001	< 0.001

组别	n	穿越平台次数
正常对照组	6	3.917±0.6994
假手术组	6	3.083±0.5128
模型组	6	1.917±0.4876^a,b
电针组	6	3.167±0.4641^c
F值		4.371
P值		0.009

组别	n	P35/P25	CDK5	Tau5
正常对照组	6	0.255±0.006	0.265±0.011	0.267±0.013
假手术组	6	0.271±0.009	0.269±0.009	0.245±0.006
模型组	6	0.346±0.017^a,b	0.369±0.016^a,b	0.274±0.012
电针组	6	0.255±0.008^c	0.270±0.006^c	0.276±0.007
F值		16.740	19.790	2.183
P值		< 0.001	< 0.001	0.122

电针对阿尔茨海默病大鼠学习记忆能力及前额叶P35/P25-周期蛋白依赖性激酶5-Tau蛋白磷酸化通路的影响

Effects of Electroacupuncture on Learning-memory and P35/P25-Cyclin-dependent Kinase 5-Tau Phosphorylation Signal Pathway in Prefrontal Cortex of Rats with Alzheimer's Disease

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组别	n	Tau5	Tau[pS199]	Tau[pS202]	CDK5	P35	P25
正常对照组	6	2.231±0.186	1.064±0.073	0.880±0.096	0.707±0.176	0.672±0.077	0.544±0.088
假手术组	6	2.351±0.173	1.217±0.068	1.218±0.067	0.882±0.126	0.8204±0.056	0.616±0.135
模型组	6	2.322±0.202	1.591±0.144^a,b	1.876±0.139^d,e	1.340±0.118^g,h	1.200±0.118^j,k	1.164±0.128^m,n
电针组	6	2.257±0.210	1.198±0.067^c	1.235±0.102^f	0.927±0.139ⁱ	0.795±0.090^l	0.696±0.038^o
F值		0.083	5.813	15.910	3.592	6.683	7.158
P值		0.969	0.005	< 0.001	0.032	0.001	0.012

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