“标本配穴”电针预处理调控心肌缺血再灌注损伤大鼠心肌细胞线粒体分裂的作用机制  

Mechanism of the pretreatment with electroacupuncture of"biaoben acupoint combination"for regulating cardiomyocyte mitochondrial fission in the rats of myocardial ischemia-reperfusion injury

作　　者：张艳琳 吴松 郭倩茹 余云涛 王孙伊 韦禹岐 万小曼 路珍 何晓茹 ZHANG Yanlin;WU Songg;GUO Qianru;YU Yuntao;WANG Sunyi;WEI Yuqi;WAN Xiaoman;LUZhen;HE Xiaoru(College of Acupuncture-Moxibustion,Orthopedics and Traumatology,Hubei University of CM/Hubei Provincial Collaborative Innovation Center of Disease Prevention with Acupuncture and Moxibustion,Wuhan 430061,China;Hubei Shizhen Laboratory,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]湖北时珍实验室,武汉430061

出　　处：《中国针灸》2025年第3期335-344,共10页Chinese Acupuncture & Moxibustion

基　　金：2022年国家中医药管理局青年岐黄学者培养项目:国中医药人教函[2022]256号;全国名老中医药专家传承工作室建设项目:国中医药人教函[2022]75号。

摘　　要：目的:观察“标本配穴”电针预处理对心肌缺血再灌注损伤(MIRI)大鼠心肌细胞线粒体分裂的影响,并探讨其作用机制。方法:将50只雄性SD大鼠随机分为假手术组、模型组、电针预处理组、电针预处理+Compound C组和电针预处理+ML385组,各10只。电针预处理组、电针预处理+Compound C组和电针预处理+ML385组于双侧“内关”“足三里”和“关元”行电针干预20 min,连续波,频率2 Hz,电流1 mA,每日1次,连续7 d。第8天,电针预处理+Compound C组和电针预处理+ML385组于造模前30 min分别腹腔注射Compound C溶液(0.3 mg/kg)和ML385溶液(30 mg/kg)。除假手术组外,其余组大鼠均采用冠状动脉左前降支结扎法建立MIRI大鼠模型,假手术组穿线不结扎。造模结束后,采用流式细胞仪检测缺血区心肌组织活性氧(ROS)含量,黄嘌呤氧化酶法检测缺血区心肌组织超氧化物歧化酶(SOD)含量,硫代巴比妥酸比色法检测缺血区心肌组织丙二醛(MDA)含量,HE染色法观察缺血区心肌组织形态,透射电镜观察缺血区心肌细胞线粒体超微结构,免疫荧光法检测缺血区心肌组织线粒体分裂因子(MFF)、线粒体裂变1蛋白抗体(Fis1)和动力蛋白相关蛋白1(Drp1)阳性表达,免疫组织化学法检测缺血区心肌组织腺苷酸活化蛋白激酶(AMPK)、核因子E2相关因子2(Nrf2)和Drp1蛋白表达。结果:与假手术组比较,模型组缺血区心肌组织ROS、MDA含量及MFF、Fis1、Drp1阳性表达均升高(P<0.01),SOD含量及AMPK、Nrf2蛋白表达均降低(P<0.01),Drp1蛋白表达升高(P<0.01)。与模型组比较,电针预处理组缺血区心肌组织ROS、MDA含量及MFF、Fis1、Drp1阳性表达均降低(P<0.01),SOD含量及AMPK、Nrf2蛋白表达均升高(P<0.01),Drp1蛋白表达降低(P<0.01);电针预处理+Compound C组和电针预处理+ML385组缺血区心肌组织MFF、Fis1、Drp1阳性表达及Drp1蛋白表达均降低(P<0.01)。与电针预处理+Compound C组和电针预处理+ML3Objective To observe the effect of electroacupuncture(EA)pretreatment of"biaoben acupoint combination"on cardiomyocyte mitochondrial fission in the rats with myocardial ischemia-reperfusion injury(MIRI)and explore its mechanism.Methods Fifty male SD rats were randomly divided into a sham-operation group,a model group,an EA pretreatment group,an EA pretreatment+Compound C group and an EA pretreatment+ML385 group,10 rats in each group.In the EA pretreatment,the EA pretreatment+Compound C group and the EA pretreatment+ML385 group,EA was delivered at bilateral"Neiguan"(PC6),"Zusanli"(ST36)and"Guanyuan"(CV4)for 20 min,with continuous wave and 2 Hz of frequency,1 mA of current,once daily for consecutive 7 days.On day 8,in the EA pretreatment+Compound C group and the EA pretreatment+ML385 group,30 min before model preparation,the intraperitoneal injection with Compound C(0.3 mg/kg)and ML385(30 mg/kg)was administered respectively.Except in the sham-operation group,the ligation of the left anterior descending coronary artery was performed to prepare MIRI rat model in the rest groups.In the sham-operation group,the thread was not ligated.After modeling,the content of reactive oxygen species(ROS)in the ischemic area was measured by flow cytometry,superoxide dismutase(SOD)was detected using xanthine oxidase method,and malondialdelyde(MDA)was detected using thiobarbituric acid(TBA)chromatometry.The morphology of myocardial tissue in the ischemic area was observed with HE staining,and the mitochondria ultrastructure of cardiomyocytes observed under transmission electron microscopy.Using immunofluorescence analysis,the positive expression of mitochondrial fission factor(MFF),mitochondrial fission 1 protein antibody(Fis1)and dynamin-related protein 1(Drp1)was detected;and with immunohistochemical method used,the protein expression of adenosine monophosphate-activated protein kinase(AMPK),nuclear factor E2-associated factor2(Nrf2)and Drp1 in the ischemic area was detected.Results Compared with the sham-operation group,the content

关 键 词：心肌缺血再灌注损伤 电针 标本配穴 预处理 线粒体分裂 AMPK Nrf2 Drp1 

分 类 号：R245[医药卫生—针灸推拿学]