针刺研究2025，Vol.50Issue(3)：251-259,9.DOI:10.13702/j.1000-0607.20241273
基于海马小胶质细胞α7烟碱乙酰胆碱受体途径探讨电针"足三里"改善系统性炎性反应小鼠认知障碍的作用机制
Mechanism of electroacupuncture at"Zusanli"(ST36)in improving cognitive impairment in mice with systemic inflammation based on the α7 nicotinic acetylcholine receptor pathway of hippocampal microglia
摘要
Abstract
Objective To observe the effect of low-intensity electroacupuncture(EA)at"Zusanli"(ST36)on the hippocampal cholinergic anti-inflammatory pathway and α7 nicotinic acetylcholine receptor(α7nAChR)of hippocampal microglia in mice with systemic inflammatory cognitive impairment,so as to investigate the central mechanism of EA in improving neuroinflammatory response and cognitive function.Methods C57BL/6 mice were randomly divided into blank,model and EA groups(6 mice/group);another set of C57BL/6 mice were randomly divided into blank,model,EA and EA+antagonist groups(6 mice/group);Cx3Cr1-Cre/ERT2 mice and α7nAChRfl/fl mice were mated to construct transgenic mice with microglial α7nAChR knockout(EA+knockout group,6 mice),with the same litter negative mice used as non-knockout mice and randomly divided into blank,model+non-knockout and EA+non-knockout groups(6 mice/group).The systemic inflammatory cognitive impairment model was established by intraperitoneal injection of lipopolysaccharide(LPS,1 mg/kg).Mice in the EA group received EA at bilateral ST36(dense-sparse wave,1 Hz/20 Hz,0.5 mA),for 30 minutes each time;mice in the EA+antagonist group received injection of α7nAChR antagonist methyllycaconitine citrate(MLA)into hippocampus.The episodic memory behavioral tests were used to observe the abilities of mice in new object exploration,object location change recognition and object temporal order change memory.Nuclear magnetic resonance spectroscopy was used to observe the choline level in the hippocampal tissue.Western blot was used to detect the protein expression of α7nAChR in the hippocampus.Immunofluorescence staining was used to detect the expression of α7nAChR in hippocampal microglia and the activation of microglia.Real-time quantitative PCR was used to detect the mRNA expression of tumor necrosis factor-α(TNF-α)in hippocampus,and ELISA was used to detect the contents of α7nAChR,inflammatory factors and microglial phenotypic markers in hippocampal tissue.Results Compared with the blank group,the abilities of new object exploration,location change recognition and object temporal order change memory,the choline level in the hippocampus,the protein expression level and content of α7nAChR and its co-expression with microglial markers,and the contents of anti-inflammatory factor interleukin(IL)-10 and M2 macrophage marker arginase 1(Arg1)of mice in the model group were decreased(P<0.001,P<0.01);while the expression of hippocampal ionized calcium-binding adapter molecule 1(Iba-1),the contents of M1 macrophage marker CD206,inducible nitric oxide synthase(iNOS),the content of pro-inflammatory factor IL-1β and the mRNA expression of TNF-α were increased(P<0.001,P<0.01).Compared with the model group,the above indexes in the EA group were reversed except for the ability of location change recognition(P<0.05,P<0.01,P<0.001).Compared with the EA group,the abilities of new object exploration and object temporal order change memory in the EA+antagonist group were decreased(P<0.05),and the mRNA expression of TNF-α was increased(P<0.01);the abilities of new object exploration and object temporal order change memory in the EA+knockout group were decreased(P<0.05),and the mRNA expression of TNF-α was increased(P<0.01).Conclusion EA at ST36 with low-intensity may improve neuroinflammatory response and systemic inflammatory cognitive dysfunction by increasing the choline level in the hippocampus and reducing the release of inflammatory factors through up-regulating the expression of α7nAChR in hippocampal microglia.关键词
电针/系统性炎性反应/认知障碍/小胶质细胞/α7烟碱乙酰胆碱受体Key words
Electroacupuncture/Systemic inflammation/Cognitive impairment/Microglia/α7nAChR引用本文复制引用
陈小梅,黄于婷,柯以晨,庞莉娜,黄秋玲,兰艳艳,俞向梅,王志福..基于海马小胶质细胞α7烟碱乙酰胆碱受体途径探讨电针"足三里"改善系统性炎性反应小鼠认知障碍的作用机制[J].针刺研究,2025,50(3):251-259,9.基金项目
国家自然科学基金面上项目(No.82274658) （No.82274658）
福建省自然科学基金项目(No.2022J01347) （No.2022J01347）
福建省卫健委医学创新项目(No.2022CXA053) （No.2022CXA053）
福建中医药大学针灸学科开放课题(No.XZJ2024002) （No.XZJ2024002）
福建省自然科学基金项目(No.2023J01333) （No.2023J01333）
福建省中青年教师教育科研项目(No.JAT220116) （No.JAT220116）
十四·五国家中医药管理局中医康复学科开放课题(No.X2023010) （No.X2023010）
福建中医药大学校管项目(No.X2023014) （No.X2023014）
