电针预处理对AMI小鼠心肌局部炎症及DNA-PK/Rictor/Myc信号通路的影响  

Effects of Electroacupuncture Preconditioning on Local Inflammation and DNA-PK/Rictor/Myc Signaling Pathway in Myocardium of Acute Myocardial Ischemia Mice

作　　者：江敏娇 彭柔 闫煜杭 刘肖儿 钱丹颖 鲁晓涵 陈栎遥 于美玲[1] 卢圣锋[1,3] JIANG Minjiao;PENG Rou;YAN Yuhang;LIU Xiaoer;QIAN Danying;LU Xiaohan;CHEN Liyao;YU Meiling;LU Shengfeng(Key Laboratory of Acupuncture and Medicine Research of Ministry of Education,Nanjing University of Chinese Medicine,Nanjing 210023,China;Zhangjiagang TCM Hospital Affiliated to Nanjing University of Chinese Medicine,Suzhou 215600,China;School of Elderly Care Services and Management,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学针药结合教育部重点实验室,江苏南京210023 [2]南京中医药大学附属张家港医院,江苏苏州215600 [3]南京中医药大学养老服务与管理学院,江苏南京210023

出　　处：《南京中医药大学学报》2024年第6期589-597,共9页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82374337,81774210);江苏省高校自然科学研究重大项目(21KJA360003);江苏高校“青蓝工程”中青年学术带头人项目(2020年);江苏省研究生科研创新计划(KYCX22-1964);张家港市科技计划项目(ZKYL2253)。

摘　　要：目的观察内关穴电针预处理后急性心肌梗死小鼠(AMI)心功能、局部炎症水平及巨噬细胞M2型极化变化,并从调控DNA-PK/Rictor/Myc信号通路角度探讨其可能机制。方法将雄性C57BL/6J小鼠随机分为假手术组、模型组和电针组,每组10只。采用结扎冠状动脉左前降支制备心肌缺血模型。电针组接受双侧内关穴电针干预,疏密波,2/15 Hz,1 mA,每次20 min,每日1次,连续3 d,电针干预结束后30 min造模。采用超声心动图检测心脏射血分数(EF)和短轴收缩率(FS);HE、TUNEL染色观察小鼠心肌病理形态和心肌细胞凋亡;免疫组化和ELISA法检测梗死区心肌组织和外周血中IL-1β、TNF-α、NLRP3的表达水平;流式细胞术检测心脏中巨噬细胞极化状态,Western blot法检测梗死心肌中DNA-PK、p-DNA-PK、Rictor、Myc的蛋白表达水平。结果与假手术组相比,模型组的EF、FS明显降低(P<0.0001),炎性细胞浸润明显,心肌细胞凋亡指数显著升高(P<0.001),心肌组织和血清中IL-1β、NLRP3、TNF-α表达上调(P<0.01,P<0.001),巨噬细胞百分比明显增加(P<0.001),M2型巨噬细胞百分比下降(P<0.0001),心肌组织中p-DNA-PK、Rictor和Myc蛋白表达水平明显降低(P<0.05,P<0.0001);与模型组相比,电针组的EF、FS显著升高(P<0.0001),炎性细胞浸润减少,心肌细胞凋亡指数下降(P<0.01),心肌组织和血清中IL-1β、NLRP3、TNF-α表达下调(P<0.05,P<0.01,P<0.001),巨噬细胞百分比下降(P<0.05),M2型巨噬细胞百分比上升(P<0.01),心肌组织中p-DNA-PK、Rictor和Myc蛋白表达显著升高(P<0.05,P<0.01,P<0.0001)。结论电针预处理可能通过调控DNA-PK/Rictor/Myc信号通路,促进巨噬细胞M2极化,减轻局部炎症,减少心肌细胞凋亡,从而提高心功能,实现心肌保护效应。OBJECTIVE To observe the changes of cardiac function,local inflammation level and macrophage M2 polarization in mice with acute myocardial infarction(AMI)after electroacupuncture preconditioning at the Neiguan point,and to explore the possible mechanisms from the perspective of regulating the DNA-PK/Rictor/Myc signaling pathway.METHODS Male C57BL/6J mice were randomly divided into sham group,model group and electroacupuncture group,with 10 mice in each group.The electroacupuncture group received bilateral electroacupuncture interventions at the Neiguan points,sparse-dense wave,2/15 Hz,1 mA,20 min/time,once a day for 3 consecutive days,and AMI models were performed 0.5 h after the electroacupuncture interventions.The myocardial ischemia model was prepared by ligating the left anterior descending branch.Echocardiography was used to detect cardiac ejection fraction(EF)and fractional shortening(FS)to evaluate cardiac function;HE and TUNEL staining were used to observe the pathological morphology of myocardium and apoptosis of cardiomyocytes,and immunohistochemistry and ELISA were used to detect IL-1β,TNF-α and NLRP3 in infarcted myocardium and peripheral blood to evaluate the level of inflammation;flow cytometry was used to detect cardiac macrophage polarization status,and Western blot method to detect the protein expression levels of DNA-PK,p-DNA-PK,Rictor and Myc in infarcted myocardium.RESULTS Compared with the sham group,the model group showed significantly lower EF and FS(P<0.0001),significant inflammatory cell infiltration,significantly higher cardiomyocyte apoptotic index(P<0.001),up-regulated expression of IL-1β,NLRP3 and TNF-α in the myocardium and serum(P<0.01,P<0.001),a significant increase in the percentage of macrophages(P<0.001),a decrease in the percentage of cardiac M2-type macrophages(P<0.0001),and a significant decrease in the expression levels of p-DNA-PK,Rictor and Myc proteins in myocardium(P<0.05,P<0.0001).Compared with the model group,EF and FS were significantly higher in the electroacupun

关 键 词：心肌梗死 电针预处理 巨噬细胞极化 DNA-PK RICTOR 

分 类 号：R285.5[医药卫生—中药学]