电针预处理对心肌缺血/再灌注损伤大鼠线粒体活性氧介导的细胞凋亡的影响  

Effect of electroacupuncture preconditioning on cell apoptosis mediated by mitochondrial reactive oxygen species in myocardial ischemia/reperfusion injury rats

作　　者：张小蕾 黄伟 杨琪琪[1,2] 向丽莉 薛艺璇 林依梦 姜功道 ZHANG Xiao-lei;HUANG Wei;YANG Qi-qi;XIANG Li-li;XUE Yi-xuan;LIN Yi-meng;JIANG Gong-dao(College of Acupuncture and Orthopedics,Hubei University of Chinese Medicine/Hubei Provincial Collaborative Innovation Center of Chinese Medicine,Wuhan 430061,China;Department of Acupuncture-moxibustion,Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430061)

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]湖北省中医院针灸科,武汉430061

出　　处：《针刺研究》2020年第12期961-967,共7页Acupuncture Research

基　　金：国家自然科学基金青年项目(No.81704142);湖北省自然科学基金项目(No.2016CFB215);武汉市中青年医学骨干人才培养工程项目(武卫生计生通[2018]116号);中国科协青年人才托举工程(No.2017QNRC001)。

摘　　要：目的:探讨电针预处理对心肌缺血再灌注损伤(MIRI)大鼠的保护作用及其对线粒体活性氧(ROS)的作用机制。方法:将80只SPF级雄性Wistar大鼠随机分为正常组、假手术组、再灌注组和电针组,每组20只。正常组、假手术组、再灌注组均给予捆绑,电针组给予电针预处理,均20 min/次,1次/d,共7 d。第8天正常组麻醉50 min后、假手术组开胸暴露心脏50 min后、再灌注组及电针组开胸缺血20 min再灌注30 min后取材。根据心电图STⅡ段变化,判断MIRI模型是否成功;采用室性心律失常(VA)评分评价心律失常情况;采用ELISA法检测血清肌酸激酶同工酶(CK-MB)含量;采用DHE染色法检测心肌组织中ROS含量;采用实时荧光定量PCR法检测心肌组织中细胞色素C(Cyt-C)、半胱天冬酶9(Caspase-9)、半胱天冬酶3(Caspase-3)基因表达水平。结果:正常组与假手术组在各项指标上差异均无统计学意义(P>0.05);与假手术组比较,再灌注组的VA评分、血清CK-MB含量、心肌ROS含量及Cyt-C、Caspase-9、Caspase-3 mRNA表达均明显升高(P<0.01);与再灌注组比较,电针组VA评分、血清CK-MB含量、心肌ROS含量及Cyt-C、Caspase-9、Caspase-3 mRNA表达均明显下降(P<0.01)。结论:电针预处理对MIRI大鼠的保护作用可能是基于调控ROS介导的凋亡通路产生的。Objective To observe the effect of electroacupuncture(EA)preconditioning on cell apoptosis and the content of mitochondrial reactive oxygen species(ROS)in rats with myocardial ischemia/reperfusion injury(MIRI),so as to explore their mechanisms underlying improvement of MIRI.Methods Eighty male Wistar rats were randomly divided into control,sham,ischemia reperfusion(IR)and EA groups,with 20 rats in each group.Rats of the control,sham and IR groups were just banded on the boards for 30 min,once daily for 7 days.Before modeling,EA(2 Hz,1 mA)was applied to"Neiguan"(PC6),"Zusanli"(ST36),"Guanyuan"(CV4)for 20 min,once daily for 7 successive days in the EA group.The MIRI model was established by occlusion of the left anterior descending branch of the left coronary artery for 20 min,followed by reperfusion for 30 min.Ventricular arrhythmia(VA)score was used to evaluate arrhythmia.Enzyme-linked immunosorbent assay(ELISA)was used to detect the level of CK-MB.DHE staining was used to detect the content of reactive oxygen species(ROS).The gene expression levels of cytochrome C(Cyt-C),Caspase-9 and Caspase-3 were detected by real-time fluorescent quantitative PCR.Results Compared with the sham group,VA score,serum CK-MB content,ROS content in heart tissue and Cyt-C,Caspase-9 and Caspase-3 gene expression were significantly up-regulated in the MIRI group(all P<0.01).Following the intervention,the increased VA score,serum CK-MB content,ROS content in heart tissue,Cyt-C,Caspase-9 and Caspase-3 gene expression were all reversed in the EA group compared with the MIRI group(all P<0.01).Conclusion The protective effect of EA preconditioning on MIRI may be based on the regulation of ROS mediated-apoptosis pathway.

关 键 词：心肌缺血再灌注损伤 电针预处理 肌酸激酶同工酶 活性氧 细胞色素C 半胱天冬酶9 半胱天冬酶3 

分 类 号：R245.97[医药卫生—针灸推拿学]