【摘要】 目的:探讨电针(electroacupuncture,EA)对脑缺血大鼠海马区microRNA132(miR132)和p250GAP蛋白表达的影响及可能内在机制。方法:将90只雄性SD大鼠随机分为假手术组、模型组、EA组、EA+H89(N-[2-(pBromocinnamylamino)ethyl]-5-isoquinolinesulfonamide·2HCl hydrate)组和EA+NS(normal saline)组。永久性结扎双侧颈总动脉(2-vessel occlusion,2VO)制备脑缺血模型,造模成功次日于百会穴(GV20)和大椎穴(GV14)施以EA治疗。治疗后的7,14,21d,采用实时荧光定量PCR(Quantitative Real-time PCR,RT-PCR)方法检测海马miR132的表达,并通过蛋白免疫印迹(western blot)方法检测海马GTP酶活化蛋白p250GAP蛋白的表达。结果:同时间点各组大鼠间相比,7、14、21d时,模型组与假手术组相比,大鼠海马组织miR132的表达量均显著下降,同时p250GAP蛋白量均显著增加(P<0.05);EA组与模型组相比,miR132的表达量均显著增加(P<0.05);同时p250GAP蛋白量均显著下调(P<0.05);EA+H89组与EA+NS组相比,miR132表达量均显著下调(P<0.05);p250GAP蛋白量均显著增加(P<0.05)。同组别三个不同时间点之间miR132表达量相比,EA+NS组大鼠在14d时miR132的表达量较7d显著增加,但21d时miR132的表达量较14d显著减少(P<0.05),其余组各时间点间无显著统计学差异。结论:EA能促进脑缺血大鼠海马区miR132增加并下调p250GAP蛋白的表达,给予H89后电针的作用被部分抑制,提示电针的作用可能与激活PKA/CREB信号通路相关。更多还原

【Abstract】 Objective:To investigate whether and how electroacupuncture(EA)affects the expression of miR132 and p250GAP protein in the hippocampal neurons of rats following cerebral ischemia.Methods:Ninety SpragueDawley(SD)rats were randomly divided into five groups:sham-operated control group,model group,EA group,EA combined with intracerebroventricular(ICV)injection of PKA blocker H89(EA+H89)group and EA combined with ICV injection of normal saline(NS)(EA+NS)group.Cerebral ischemia was induced by permanent,bilateral common carotid artery occlusion(2-vessel occlusion,2VO),and EA was delivered the day after the operation at Baihui(GV20)and Dazhui(GV14)acupoints.Quantitative real-time PCR(RT-PCR)and Western blotting were respectively employed to detect the changes of miR132 and p250GAP protein in the hippocampus on the day 7,14or21 after the EA treatment.Results:On the day 7,14 and 21,the expression levels of miR132 were significantly decreased and the p250 GAP protein were significantly increased in the model group as compared with the control group(P<0.05),while those of miR132 were significantly higher and those of p250 GAP protein were significantly lower in the EA group than in the model group(P<0.05).In the EA+H89group,protective effects of EA were weakened,whereas in the EA+NS group they were not affected.Among the 3time points in each group,the expression of miR132 in the EA+NS groups was significantly higher on the day 14 than that on the day 7(P<0.05),but that was significantly lower on the day 21 than that on the day 14(P<0.05).No statistically significant difference was found among the other groups at each time point.Conclusion:EA could promote the increaseof miR132 and inhibit the decrease of p250 GAP protein in the hippocampus of cerebral ischemia rats,and these effects could be inhibited by H89,suggesting that PKA/CREB signaling pathway might be activated in effects of EA.更多还原