【摘要】 目的探讨经皮穴位电刺激(TEAS)对行腹部手术老年大鼠的脑保护作用及其可能的作用机制。方法取35只大鼠行腹腔探查术+脾切除术,将30只术后无行动障碍的大鼠随机分为手术组、TEAS组和西药组各10只;另取10只大鼠作为对照组,仅腹腔注射生理盐水。TEAS组选取特定穴位给予电刺激和生理盐水灌胃,西药组给予非特定穴位电刺激和石杉碱灌胃,手术组和对照组给予非特定穴位电刺激和生理盐水灌胃。末次治疗后12 h,利用Morris水迷宫实验检测各组大鼠的学习记忆能力,旷场实验检测大鼠的行为学改变;处死大鼠后取海马组织,苏木精-伊红染色观察海马组织病理学变化,TUNEL染色观察海马神经元凋亡率,透射电镜观察海马自噬情况,蛋白免疫印迹法检测海马组织腺苷酸活化蛋白激酶(AMPKα)、磷酸化AMPKα(p-AMPKα)、Beclin1、液泡分选蛋白34(Vps34)、微管相关蛋白1轻链3(LC3)蛋白表达,比较各组上述指标的差异。结果与对照组比较,其他3组的逃避潜伏期延长,原平台象限停留时间缩短,穿越平台次数、水平运动得分、垂直运动得分减少,海马组织神经元细胞形态发生改变,神经元细胞凋亡率及自噬体均增加,LC3-Ⅱ/LC3-Ⅰ比值和海马组织p-AMPKα、Beclin1、Vps34的蛋白表达升高(均P<0.05);TEAS组和西药组以上指标均较手术组有所改善(均P<0.05),而TEAS组和西药组之间差异并无统计学意义(均P>0.05)。结论 TEAS可改善行腹部手术的老年大鼠的认知功能障碍,减少神经元细胞凋亡和自噬,具有与常规西药相似的脑保护作用,其可能是通过抑制AMPK-Beclin1-Vps34通路发挥作用。更多还原

【Abstract】 Objective To explore the brain protective effect of transcutaneous electrical acupoint stimulation(TEAS) on aged rats undergoing abdominal surgery and its possible action mechanism.Methods Thirty-five rats underwent abdominal exploration and splenectomy, wherein 30 rats without impaired mobility after operation were randomly divided into operation group(n=10), TEAS group(n=10) and western medicine group(n=10). Another 10 rats were selected as control group, and were intraperitoneally injected with normal saline only. Electrical stimulation at specific acupoints combined with intragastric administration of normal saline was performed on the TEAS group, electrical stimulation at non-specific acupoints combined with intragastric administration of huperzine on the western medicine group, and electrical stimulation at non-specific acupoints combined with intragastric administration of normal saline on the operation group and control group. Twelve hours after last administration, Morris water maze was used to detect the learning and memory ability of rats in each group, and open field test was employed to detect changes in rat behavior. The rats were sacrificed to harvest their hippocampus tissues, then the histopathological changes in hippocampus tissues were observed after hematoxylin-eosin staining, hippocampal neuronal apoptosis rate was determined by TUNEL staining, hippocampal autophagy was observed under transmission electron microscope, and the protein expression of adenosine monophosphate-activated protein kinase α(AMPKα), phosphorylated AMPKα(p-AMPKα), Beclin1, vacuolar protein sorting 34(Vps34), and microtubule-associated protein 1 light chain 3(LC3) in hippocampus tissues was detected by Western blotting. Differences in the aforementioned indicators were compared among groups.Results Compared with the control group, the other three groups possessed longer escape latency, shorter length of original platform quadrant stay, less frequency of crossing platforms, lower scores of horizontal and vertical movement, changes in hippocampal neuron morphology, increased neuronal cell apoptosis rate and autophagosome number, as well as higher LC3-Ⅱ/LC3-Ⅰ ratio and elevated expression of p-AMPKα, Beclin1 and Vps34 proteins in hippocampus tissues(all P<0.05). The indicators mentioned above were improved in the TEAS and western medicine groups than in the operation group(all P<0.05), whereas there was no statistically significance between the TEAS group and the western medicine group(all P>0.05).Conclusion TEAS can improve cognitive dysfunction of aged rats undergoing abdominal surgery, reduce neuronal cell apoptosis and autophagy, and has similar brain protection as routine western medicine, which may exert an effect by inhibiting the AMPK-Beclin1-Vps34 pathway.更多还原