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[ RhoA/ROCK通路参与三焦针刺对阿尔茨海默病小鼠学习记忆及突触可塑性的影响 ]

[RhoA/ROCK pathway involved in effects of Sanjiao acupuncture on learning and memory and synaptic plasticity in Alzheimer's disease mice].

作者信息

Wang Yu, Zhao Lan, Shi Hui-Yan, Jia Yu-Jie, Kan Bo-Hong

机构信息

Department of Acupuncture and Moxibustion, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China.

Research Institute of Acupuncture and Moxibustion, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300193.

出版信息

Zhen Ci Yan Jiu. 2021 Aug 25;46(8):635-41. doi: 10.13702/j.1000-0607.200985.

Abstract

OBJECTIVE

To observe the effect of Sanjiao acupuncture(triple energizer acupuncture)on the small G protein guanosine triphosphate enzyme subfamily protein RhoA/Rho kinase (ROCK) pathway in Alzheimer's disease mice, and explore its effect on learning and memory function and neurosynaptic plasticity.

METHODS

Forty SAMP8 senile dementia mice were randomly divided into model, Sanjiao acupuncture (acupuncture), non acupoint acupuncture (non-acupoint) and fasudil groups, with 10 mice in each group, another 10 SAMR normal aging mice were selected as normal aging group. Mice in the acupuncture group were treated with acupuncture intervention on "Danzhong"(CV18), "Zhongwan"(CV13), "Qihai"(BL24) and bilateral "Xuehai"(SP10) and "Zusanli" (ST36). Mice in the non-acupoint group were treated with acupuncture at each of the left and right non-acupoints under the ribs and mice in the fasudil group were intraperitoneally injected with fasudil (25 mg/kg). The mice in each group were given medicine or acupuncture on the second day after grouping for 28 continuously days, once a day. Morris water maze test was used to test the learning and memory ability of mice. HE staining was used to observe the pathological changes of neurons in hippocampus. The number of hippocampal neuron dendritic spine was detected by FD fast Golgi staining kit. The contents of β-amyloid 42 (Aβ42) and phosphorylated tau protein (p-tau) in hippocampus were detected by ELISA. Western blot was used to detect the protein relative expression levels of RhoA, ROCK, F-actin and p-cofilin in hippocampus.

RESULTS

Compared with those in the normal aging group, the hippocampal neurons of the model group were disorderly arranged, the number of neuron was reduced, the escape latency, hippocampal Aβ42 and p-tau contents, RhoA and ROCK protein expressions increased (<0.05), the number of crossing the original platform, the number of neuronal dendritic spines, expressions of F-actin and p-cofilin decreased (<0.05). After the interventions, there was no statistically significant difference in the above indicators in the non-acupoint group relevant to the model group (>0.05). The acupuncture group and fasudil group improved the hippocampal pathological damage. The escape latency, hippocampal Aβ42 and p-tau contents, the expressions of RhoA and ROCK protein decreased (<0.05), and the number of crossing the original platform, the number of hippocampal neuron dendritic spines, expressions of F-actin and p-cofilin increased (<0.05) in both of the acupuncture and fasudil groups in contrast to the model and non-acupoint groups. Compared with the acupuncture group, there was no significant difference in the above indicators in the fasudil group (>0.05).

CONCLUSION

Sanjiao acupuncture may inhibit the activation of the RhoA/ROCK pathway, so as to improve the learning and memory function of AD mice, increase the number of hippocampal neuron dendritic spines, and promote synaptic plasticity.

摘要

目的

观察三焦针法对阿尔茨海默病小鼠小G蛋白鸟苷三磷酸酶亚家族蛋白RhoA/Rho激酶(ROCK)通路的影响,探讨其对学习记忆功能及神经突触可塑性的作用。

方法

将40只SAMP8老年痴呆小鼠随机分为模型组、三焦针法组(针法组)、非经非穴针法组(非穴组)和法舒地尔组,每组10只,另选10只SAMR正常衰老小鼠作为正常衰老组。针法组小鼠取“膻中”(CV18)、“中脘”(CV13)、“气海”(BL24)及双侧“血海”(SP10)、“足三里”(ST36)进行针刺干预。非穴组小鼠取双侧肋下非经非穴点针刺,法舒地尔组小鼠腹腔注射法舒地尔(25 mg/kg)。各组小鼠于分组后第2天开始给药或针刺,连续28天,每日1次。采用Morris水迷宫试验检测小鼠学习记忆能力。采用HE染色观察海马神经元病理变化。采用FD快速高尔基染色试剂盒检测海马神经元树突棘数量。采用ELISA法检测海马中β淀粉样蛋白42(Aβ42)和磷酸化tau蛋白(p-tau)含量。采用Western blot法检测海马中RhoA、ROCK、F-肌动蛋白和p-丝切蛋白的蛋白相对表达水平。

结果

与正常衰老组相比,模型组海马神经元排列紊乱,神经元数量减少,逃避潜伏期、海马Aβ42和p-tau含量、RhoA和ROCK蛋白表达增加(P<0.05),穿越原平台次数、神经元树突棘数量、F-肌动蛋白和p-丝切蛋白表达减少(P<0.05)。干预后,非穴组上述指标与模型组比较,差异无统计学意义(P>0.05)。针法组和法舒地尔组改善了海马病理损伤。与模型组和非穴组比较,针法组和法舒地尔组逃避潜伏期、海马Aβ42和p-tau含量、RhoA和ROCK蛋白表达降低(P<0.05),穿越原平台次数、海马神经元树突棘数量、F-肌动蛋白和p-丝切蛋白表达增加(P<0.05)。与针法组比较,法舒地尔组上述指标差异无统计学意义(P>0.05)。

结论

三焦针法可能通过抑制RhoA/ROCK通路的激活,从而改善AD小鼠的学习记忆功能,增加海马神经元树突棘数量,促进突触可塑性。

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