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目的:观察电针预处理对大鼠脑缺血再灌注后小胶质细胞活化状态的影响并探讨其机制。方法:成年雄性SD大鼠随机分为7组,分别为假手术组(Sham),脑缺血再灌注组(MCAO),电针(EA)+MCAO组,烟碱型乙酰胆碱受体α7亚单位(α7nAChR)PHA-543,613+MCAO组,溶剂(Vehicle)+MCAO组,α7nAChR抑制剂α-BGT+EA+MCAO组,溶剂+EA+MCAO组。脑缺血再灌注后3 d取材,运用Western Blot技术以及免疫荧光技术检测脑缺血半暗带小胶质细胞经典激活状态(M1型)及选择性激活状态(M2型)特异性标志分子的表达水平。结果:(1)与MCAO组比较,电针预处理组(EA+MCAO)M1型小胶质细胞标志分子iNOS表达下调(P<0.05),M2型小胶质细胞标志分子Arginase表达上调(P<0.05);(2)α7nAChR激动剂PHA-543,613可模拟电针预处理的作用,使小胶质细胞由M1型向M2型转化;α7nAChR抑制剂α-BGT逆转了电针促进小胶质细胞由M1型向M2型转化的作用。结论:电针预处理可以使脑缺血再灌注后小胶质细胞由M1型向M2型转化,α7nAChR参与了电针预处理对小胶质细胞活化状态改变的过程。
Objective: To observe the effect of electroacupuncture (EA) preconditioning on the activation of microglia after cerebral ischemia-reperfusion in rats and its mechanism. Methods: Adult male Sprague-Dawley rats were randomly divided into 7 groups: Sham, MCAO, EA + MCAO, nicotinic acetylcholine receptor α7 subunit (α7nAChR) PHA-543,613 + MCAO group, vehicle + MCAO group, α7nAChR inhibitor α-BGT + EA + MCAO group and solvent + EA + MCAO group. At 3 days after cerebral ischemia / reperfusion, Western Blot and immunofluorescence were used to detect the expression of specific markers (M1) and selective activation (M2) in microglia of ischemic penumbra The expression level. Results: (1) Compared with MCAO group, the iNOS expression of M1 microglial marker in EA + MCAO group was down-regulated (P <0.05), and the expression of Arginase in M2 microglia marker was up-regulated <0.05). (2) α7nAChR agonist PHA-543,613 could mimic the effect of electroacupuncture preconditioning and make microglia transform from M1 type to M2 type. Alpha7nAChR inhibitor α-BGT reversed the effect of EA on microglial cells The role of conversion from M1 to M2. CONCLUSION: Electroacupuncture preconditioning can change microglial cells from M1 type to M2 type after cerebral ischemia-reperfusion. Α7 nAChR is involved in the process of microglial activation induced by EA preconditioning.