针刺研究

2020, v.45(07) 529-534

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电针对SAMP8小鼠海马区细胞周期蛋白依赖性激酶5及Tau蛋白的影响
Effect of electroacupuncture on cyclin-dependent kinase 5 and Tau protein in hippocampus of SAMP8 mice

马冉;孔立红;齐凤军;何瑞阳;郑清;卢威;付晓平;
MA Ran;KONG Li-hong;QI Feng-jun;HE Rui-yang;ZHENG Qing;LU Wei;FU Xiao-ping;College of Acupuncture and Orthopedics,Hubei University of Chinese Medicine;Guanggu Branch of Hubei Hospital of Traditional Chinese Medicine/Hubei Province Academy of Traditional Chinese Medicine;

摘要(Abstract):

目的:观察电针"肾俞"对SAMP8小鼠海马区神经元超微结构及细胞周期蛋白依赖性激酶5(CDK5)、Tau蛋白的影响,探讨电针防治阿尔茨海默病(AD)的作用机制。方法:将5月龄雄性SAMP8小鼠随机分为模型组、电针组和针刺组,以同品系SAMR1小鼠为正常组,每组15只。针刺组针刺小鼠"百会"及双侧"肾俞";电针组针刺"百会"并电针双侧"肾俞"。两组均20 min/次,每日1次,7 d为1个疗程,共治疗4个疗程。治疗后行Morris水迷宫检测小鼠的学习记忆能力,透射电镜观察海马区超微结构,免疫组织化学法检测海马CDK5的阳性表达,Western blot法检测海马区Tau-5和p25、CDK5蛋白的表达水平。结果:①水迷宫测试结果显示:与正常组比较,模型组小鼠平均逃避潜伏期延长(P<0.05,P<0.01),原平台象限停留时间缩短、跨越原平台次数减少(P<0.01);与模型组比较,电针组和针刺组平均逃避潜伏期缩短(P<0.05,P<0.01),电针组原平台象限停留时间延长、跨越原平台次数增多(P<0.01),针刺组原平台象限停留时间延长(P<0.05);与电针组比较,针刺组平均逃避潜伏期延长(P<0.05),原平台象限停留时间缩短(P<0.05)。②透射电镜结果显示:模型组小鼠海马形态不规则且模糊不清,神经元体积缩小,线粒体数目减少,肿胀并伴随线粒体嵴畸形样改变;针刺组海马区神经元形态较规则,细胞器数目有所减少,胞质水肿严重,胞核固缩,内质网、核糖体及线粒体未见明显异常;电针组神经元突触形态完整,核和膜结构清晰,可见较多数量的细胞器,内质网、线粒体、溶酶体和高尔基体等形态均正常。③与正常组比较,模型组海马区Tau-5和p25、CDK5蛋白相对表达量及CDK5阳性表达升高(P<0.01,P<0.05);与模型组比较,电针组和针刺组Tau-5和p25、CDK5蛋白相对表达量下降(P<0.05,P<0.01),电针组CDK5阳性表达下降(P<0.01);与电针组比较,针刺组p25和CDK5蛋白相对表达量升高(P<0.05)。结论:电针"肾俞"能降低CDK5的表达,抑制Tau蛋白,对AD的防治具有积极作用。
Objective To observe the effect of electroacupuncture(EA) and manual acupuncture(MA) on learning-memory ability, changes of ultrastructure of neurons and expression of CDK5 and Tau proteins in hippocampus of SAMP8 mice,so as to explore its mechanisms underlying improvement of Alzheimer's disease(AD).Methods A total of 45 male SAMP8 mice were randomly divided into model, EA and MA groups, with 15 mice in each group. The other 15 SAMR1 mice were used as the normal group. In the EA group, EA(2 Hz, 1 mA) was applied to bilateral "Shenshu"(BL23) and manual acupuncture was applied to "Baihui"(GV20) for 20 min. In the MA group, MA was applied to GV20 and bilateral BL23 for 20 min. Both group were treated once a day for 31 days, and with an interval of one day between every two 7 days. Morris water maze was performed to assess the animals' learning-memory ability. The morphological changes of hippocampal neurons were observed under transmission electron microscopy. The expression levels of CDK5, p25 and Tau-5 proteins in the hippocampus were detected by immunohistochemistry and Western blot, separately.Results ①Compared with the normal group, the average escape latency of Morris water maze test was prolonged in the model group(P<0.05, P<0.01), duration of swimming in the original platform quadrant and the number of original platform crossing were significantly shorter and less respectively(P<0.01). Compared with the model group, the average escape latency in the EA group was shortened(P<0.05, P <0.01), the duration of swimming in the original platform quadrant and the number of original platform crossing were significantly prolonged and increased(P<0.01); The average escape latency in the MA group was shortened(P<0.05, P <0.01),and the duration of swimming in the original platform quadrant was prolonged(P<0.05). Compared with the EA group, the average escape latency of the MA group was prolonged(P<0.05), the duration of swimming in the original platform quadrant was shortened(P<0.05). ②Transmission electron microscopy revealed that the neurons in the hippocampal CA1 area had irregular shape and vague structure, reduction in size and number of mitochondria accompanied with swelling, and malformed changes of mitochondrial crest in the model group, which was relatively milder in both EA and MA groups. ③The expression levels of hippocampal Tau-5, p25 and CDK5 proteins were significantly up-regulated in the model group in contrast to the normal group(P<0.01, P<0.05), and obviously down-regulated in both EA and MA groups relevant to the model group(P<0.05, P<0.01). Compared with the EA group, the expression levels of p25 and CDK5 proteins were significantly increased in the MA group(P<0.05). Conclusion EA of BL23 can improve the learning-memory ability in SAMP8 mice, which is associated with its effect in down-regulating the expression of hippocampal CDK5, p25 and Tau-5 proteins.

关键词(KeyWords): 阿尔茨海默病;电针;学习记忆能力;细胞周期蛋白依赖性激酶5;Tau蛋白;海马
Alzheimer's disease;Electroacupuncture;Learning-memory ability;Cyclin dependent kinase 5;Tau protein;Hippocampus

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(No.81373741)

作者(Author): 马冉;孔立红;齐凤军;何瑞阳;郑清;卢威;付晓平;
MA Ran;KONG Li-hong;QI Feng-jun;HE Rui-yang;ZHENG Qing;LU Wei;FU Xiao-ping;College of Acupuncture and Orthopedics,Hubei University of Chinese Medicine;Guanggu Branch of Hubei Hospital of Traditional Chinese Medicine/Hubei Province Academy of Traditional Chinese Medicine;

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DOI: 10.13702/j.1000-0607.190752

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