电针对脑缺血大鼠海马神经元microRNA132和p250GAP蛋白表达的影响  

Effects of electroacupuncture on expression of microRNA132 and p250GAP protein in hippocampus of cerebral ischemia rats

作　　者：郑彩霞[1] 黄晓琳[1] 陆敏[1] 张凤霞[1] 郭雅碧 韩肖华[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院康复医学科,武汉430030

出　　处：《中国康复》2016年第6期422-425,共4页Chinese Journal of Rehabilitation

基　　金：国家自然科学基金项目(NO81171858)

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。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 Sprague-Dawley （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 com-bined with ICV injection of normal saline （NS）（EA＋NS）group.Cerebral ischemia was induced by permanent,bi-lateral 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 7,14 or 21 after the EA treatment.Results：On the day 7,14 and 21,the expression levels of miR132 were significantly de-creased and the p250GAP 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 p250GAP protein were significantly lower in the EA group than in the model group （P 〈0.05）.In the EA＋H89 group,protective effects of EA were weak-ened,whereas in the EA＋NS group they were not affected.Among the 3 time points in each group,the expressi-on 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 increase＆amp;nbsp;of miR132 and inhibit the decrease of p250GAP 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.

关 键 词：脑缺血 电针 microRNA132 PKA/CREB 信号通路 

分 类 号：R245.3[医药卫生—针灸推拿学]