电针改善血管性痴呆大鼠认知障碍的机制研究
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国家自然科学基金项目(82074513,82305356);国家中医药管理局高水平中医药重点学科建设项目(zyyzdxk-2023102)——中医康复学;福建省自然科学基金计划项目(2020J01762)


Mechanism of Electroacupuncture in Improving Cognitive Impairment in Vascular Dementia Rats
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    摘要:

    目的:观察电针神庭、百会穴对血管性痴呆(VD)大鼠的影响并探讨其改善VD大鼠认知障碍的可能机制。方法:无特定病原体(SPF)级斯泼累格·多雷(SD)大鼠60只,随机选取12只作为假手术组,其余大鼠均通过双侧颈总动脉闭塞法(2-VO)构建VD模型。将造模成功的32只大鼠随机分为模型组、电针组、非穴组和电针+AMP活化的蛋白激酶(AMPK)抑制剂组(n=8)。电针组选取神庭、百会穴,非穴组选择双肋下髂嵴上10 mm进行电针干预,30 min/次,1次/d,共干预7 d;电针+AMPK抑制剂组在电针前2 h腹腔注射Compound C溶液(20 mg/kg);假手术组和模型组仅捆绑。干预结束后,使用Morris水迷宫观察大鼠的学习记忆功能;酶联免疫吸附试验(ELISA)检测血清白细胞介素-1β(IL-1β),海马组织中炎症介质白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量;蛋白质免疫印迹法和实时荧光定量PCR法分别检测海马组织中AMPKα1、核因子κB(NF-κB)p65蛋白水平和mRNA表达。结果:与假手术组比较,模型组大鼠学习、记忆功能下降;血清IL-1β与海马组织IL-6、TNF-α表达增加;AMPKα1蛋白和mRNA表达减少,NF-κB p65蛋白和mRNA表达增加(均P<0.05)。与模型组比较,电针组大鼠学习、记忆功能改善;炎症介质IL-1β、IL-6和TNF-α含量减少;AMPKα1蛋白和mRNA表达增加,NF-κB p65蛋白和mRNA表达减少(均P<0.05)。与电针组比较,非穴组和电针+AMPK抑制剂组削弱了对VD大鼠认知功能改善能力及对炎症介质和AMPK/NF-κB通路的调控作用。结论:电针神庭、百会穴能够改善VD大鼠认知功能障碍,可能与调节AMPK/NF-κB信号通路,减轻外周及海马神经炎症相关。

    Abstract:

    To observe the effects of electroacupuncture at Shenting(GV 24) and Baihui(GV 20) on vascular dementia(VD) rats and explore its potential mechanism in improving cognitive impairment in VD rats.Methods:Sixty specific-pathogen-free(SPF) Sprague-Dawley(SD) rats were randomly selected,with 12 rats as the sham surgery group.The remaining rats were subjected to bilateral common carotid artery occlusion(2-VO) to establish the VD model.Thirty-two successful model rats were randomly divided into four groups:model group,electroacupuncture group,non-acupoint group,and electroacupuncture+AMP-activated protein kinase(AMPK) inhibitor group(n=8 per group).The electroacupuncture group received electroacupuncture at Shenting(GV 24) and Baihui(GV 20),while the non-acupoint group received electroacupuncture at a site 10 mm above the iliac crest.The interventions were carried out for 30 min per session,once daily for 7 days.The electroacupuncture+AMPK inhibitor group received a 20 mg/kg Compound C solution by intraperitoneal injection 2 hours before electroacupuncture.The sham surgery group and model group only underwent binding.After the intervention,the Morris water maze test was used to observe learning and memory function.Enzyme-linked immunosorbent assay(ELISA) was used to detect serum interleukin-1β(IL-1β),and hippocampal levels of inflammatory mediators IL-6 and tumor necrosis factor-α(TNF-α).Western blotting and real-time quantitative PCR were used to detect the protein and mRNA expression levels of AMPKα1 and nuclear factor-κB(NF-κB) p65 in the hippocampus.Results:Compared with the sham surgery group,the model group exhibited decreased learning and memory function,increased serum IL-1β,and elevated hippocampal IL-6 and TNF-α levels.AMPKα1 protein and mRNA expression was reduced,while NF-κB p65 protein and mRNA expression was increased(all P<0.05).Compared with the model group,the electroacupuncture group showed improved learning and memory function,decreased levels of inflammatory mediators IL-1β,IL-6,and TNF-α,increased AMPKα1 protein and mRNA expression,and decreased NF-κB p65 protein and mRNA expression(all P<0.05).Compared with the electroacupuncture group,the non-acupoint group and electroacupuncture+AMPK inhibitor group showed reduced cognitive function improvement and weakened regulation of inflammatory mediators and the AMPK/NF-κB pathway.Conclusion:Electroacupuncture at Shenting(GV 24) and Baihui(GV 20) can improve cognitive impairment in VD rats,possibly by regulating the AMPK/NF-κB signaling pathway and alleviating peripheral and hippocampal neuroinflammation.

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唐文静,李惠菁,游咏梅,杨珊莉,王莹,李涓,饶婷,罗霄,龚一萌,钟冬灵,江一静.电针改善血管性痴呆大鼠认知障碍的机制研究[J].世界中医药,2025,(04).

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  • 收稿日期:2024-07-24
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  • 在线发布日期: 2025-05-09
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