百会及印堂穴对七氟醚麻醉后大鼠认知功能的影响及机制探究

Study on Effects of Baihui(GV 20)and Yintang(EX-HN3)on Cognitive Function of Rats After Sevoflurane Anesthesia and Its Mechanism

目的:观察电针百会穴、印堂穴对七氟醚麻醉后大鼠认知功能的影响,同时检测脑组Sonic Hedgehog(Shh)通路标志性蛋白水平的变化。方法:纳入30只SPF级SD大鼠,随机分为空白组、麻醉组及电针组,每组10只。空白组大仅进行模拟捉拿,麻醉组大鼠吸入3.0%七氟醚30 min,随后用1.5%七氟醚麻醉维持3 h,电针组在此基础上进行电针干预,连续干预7 d。干预结束后用Morris水迷宫检测各组学习记忆能力变化,HE染色法观察各组海马组织的结构变化,Western blot及Real-Time PCR法检测各组海马组织Shh、Gli、PI3K、pAKT蛋白及mRNA的水平变化。结果:1)Morris水迷宫显示:麻醉组及电针组大鼠游泳总路程及逃避潜伏期明显较空白组延长(P<0.05),其中电针组较麻醉组明显改善(P<0.05)。2)HE染色提示空白组大鼠海马组织结构形态正常,麻醉组及电针组大鼠海马组织均出现明显结构破坏,细胞排列错乱,神经元细胞出现明显变形缩小,其中电针组海马形态结构较麻醉组改善。3)Western blot及PT-PCR的结果显示,麻醉组及电针组大鼠海马组织Shh、Gli、PI3K、p-AKT蛋白及mRNA表达较空白组降低(P<0.05),其中电针组较麻醉组改善(P<0.05)。结论:七氟醚可对认知能力产生一定损害效应,电针百会穴、印堂穴可明显提升认知能力,其作用机制可能与激活Shh通路通路有关。

Objective:To observe effects of electroacupuncture at Baihui(GV 20)and Yintang(EX-HN3)on cognitive function after sevoflurane anesthesia in rats,and to detect level changes of Shh pathway marker protein in brain tissue.Methods:Thirty SPF SD rats were included in this study and randomly divided into blank group(n=10),anesthesia group(n=10)and electroacupuncture group(n=10).The rats in the blank group were only subjected to simulated capture.The rats in the anesthesia group inhaled 3.0%sevoflurane for 30 min,then they were anesthetized with 1.5%sevoflurane for 3 h.On this basis,the electroacupuncture group was treated with electroacupuncture for 7 d.After the intervention,Morris water maze was used to detect changes of learning and memory ability in each group.HE staining was used to observe structural changes of hippocampal tissue in each group.Western blot and Real-Time PCR were used to detect level changes of Shh,Gli,PI3K,pAKT protein and mRNA in hippocampal tissue in each group.Results:The Morris water maze showed that total swimming distance and escape latency in the anesthesia group and electroacupuncture group were significantly longer than those in the blank group(P<0.05),and those in the electroacupuncture group were significantly improved compared with those in the anesthesia group(P<0.05).2)HE staining showed that the morphology of hippocampal tissue was normal in the blank group,and the hippocampal tissue in the anesthesia group and electroacupuncture group showed obvious structural destruction,disordered arrangement of cells and obvious deformation and reduction of neuron cells.The morphological structure of hippocampus in the electroacupuncture group was better than that in the anesthesia group.3)Western blot and PT-PCR showed that the expressions of Shh,Gli,PI3K,p-AKT protein and mRNA in hippocampus of rats in the anesthesia group and electroacupuncture group were lower than those in the blank group(P<0.05),and those in electroacupuncture group were better than those in the anesthesia group(P<0.05).Conclusion:Sevoflurane can have a certain damage effect on cognitive ability.Electroacupuncture at Baihui(GV 20)and Yintang(EX-HN3)can obviously improve cognitive ability,and its mechanism may be related to activation of Shh pathway.