电针不同腧穴对心肌缺血模型大鼠细胞凋亡及心肌细胞miRNAs表达的影响  

Effects of electroacupuncture at different acupoints on apoptosis and the expression of miRNAs in myocardial cells in rats model of myocardial ischemia

作　　者：王华[1] 卢继东[1] 吴松[1] 杨书侠[2] 望庐山[1] 周焕娇[1] 付伊萌 刘建民[1] 

机构地区：[1]湖北中医药大学针灸骨伤学院/针灸治未病湖北省协同创新中心,武汉430061 [2]浙江农林大学健康管理系

出　　处：《中国针灸》2016年第3期281-286,共6页Chinese Acupuncture & Moxibustion

基　　金：国家自然科学基金面上项目:81473789

摘　　要：目的:观察电针不同组腧穴对心肌缺血大鼠细胞凋亡相关血清、微小核糖核酸(microRNA,miRNA)表达的影响,探讨其作用机制。方法:48只Wistar雄性大鼠随机分为正常组、模型组、内关电针组(内关组)及标本配穴电针组(配穴组),每组12只。模型组、内关组和配穴组大鼠以盐酸异丙肾上腺素(isoprenaline hydrochloride,ISO)2mg/(kg·d)背部皮下注射14d的方法制备心肌缺血模型;正常组注射等量0.9%NaCl溶液。造模后,内关组选取"内关",配穴组选取"关元""足三里""内关"分别进行电针治疗;正常组、模型组大鼠只抓取固定,每天1次,共21d。采用酶联免疫吸附试验(ELISA)检测各组大鼠血清肌酸激酶同工酶(creatine phosphate enzyme isozyme,CK-MB)、血管内皮细胞黏附分子-1(vascular cell adhesion molecule,VCAM-1)及内皮素1(ET-1)的含量;DNA断裂的原位末端标记法(TUNEL法)检测心肌细胞凋亡指数(apoptosisindex,AI);实时定量荧光PCR法检测心肌细胞miRNA-1、miRNA-133、miRNA-208、miRNA-499的表达量。结果:与正常组比较,模型组、内关组、配穴组大鼠血清CK-MB、VCAM-1、ET-1升高(均P<0.01),心肌细胞凋亡指数增多(均P<0.01);内关组、配穴组CK-MB、VCAM-1、ET-1均明显低于模型组(均P<0.01),心肌细胞凋亡指数下降(均P<0.01),且配穴组低于内关组(P<0.05)。与正常组比较,模型组心肌细胞miRNA-133表达降低(P<0.01),miRNA-208、miRNA-1、miRNA-499表达升高(均P<0.01);与模型组比较,内关组、配穴组心肌细胞miRNA-133表达增加(均P<0.01),miRNA-208、miRNA-1、miRNA-499表达降低(均P<0.01);与内关组比较,配穴组miRNA-133表达增加(P<0.01),miRNA-208、miRNA-1、miRNA-499表达降低(P<0.01,P<0.05)。结论:电针腧穴尤其是配穴组可有效防治心肌缺血,其保护机制可能与上调miRNA-133的表达,抑制心肌miRNA-1、miRNA-208、miRNA-499表达的双重调控作用有关。Objective To observe the effects of electroacupuncture（EA）at different acupoints on apoptosis-related serum and expression of microRNA（miRNA）in rats with myocardial ischemia,so as to explore its mechanism of action.Methods A total of 48 male Wistar rats were randomly divided into a normal group,a model group,a Neiguan group and a acupoint compatibility group,12 rats in each group.Isoprenaline hydrochloride（ISO）with a daily dose of 2mg/kg was subcutaneously injected for 14 days to establish the myocardial ischemia model in the model group,Neiguan group and acupoint compatibility group.Rats in the normal group were subcutaneously injected with an equal volume of normal saline.After modeling,rats in the Neiguan group were treated with EA at＂Neiguan＂（PC 6）,while rats in the acupoint compatibility group were treated with EA at＂Guanyuan＂（CV 4）,＂Zusanli＂（ST 36）and＂Neiguan＂（PC 6）.Rats in the normal group and model group were treated with immobilization,once a day for 21 days.The contents of creatine kinase-MB（CK-MB）,vascular cell adhesion molecule-1（VCAM-1）and endothelin-1（ET-1）in serum were detected by enzyme-linked immunosorbent assay（ELISA）;apoptosis index（AI）of myocardial cells was detected by TUNEL method;the expressions of miRNA-1,miRNA-133,miRNA-208 and miRNA-499 were detected by real-time PCR method.Results Compared with the normal group,the serum CK-MB,VCAM-1and ET-1were significantly increased in the model group,Neiguan group and acupoint compatibility group（all P〈0.01）,and the apoptosis index was significantly increased（all P〈0.01）.The CK-MB,VCAM-1and ET-1in the Neiguan group and acupoint compatibility group were significantly lower than those in the model group（all P〈0.01）;the AI was reduced,which was more significant in the acupoint compatibility group（P〈0.05）.Compared with the normal group,the expression of miRNA-133 was reduced（P〈0.01）and those of miRNA-208,miRNA-1and miRNA-499 were significantly increased in the

关 键 词：心肌缺血 凋亡 电针 腧穴配伍 穴 内关 穴 关元 穴 足三里 微小核糖核酸 

分 类 号：R246.1[医药卫生—针灸推拿学]