基于SLC7A11/GPX4通路探讨电针预处理对心肌缺血再灌注损伤大鼠心肌细胞铁死亡的影响  

Effect of EA Pretreatment on Ferroptosis of Myocardial Cells in Rats with MIRI Based on SLC7A11/GPX4 Pathway

作　　者：张艳琳 吴松[1,2,3,4] 路珍 韦禹岐 郭倩茹 ZHANG Yanlin;WU Song;LU Zhen;WEI Yuqi;GUO Qianru(Hubei University of Chinese Medicine,Wuhan 430061,China;Hubei Provincial Collaborative Innovation Center of Preventive Treatment by Acupuncture and Moxibustion,Wuhan 430061,China;Hubei Shizhen Laboratory,Wuhan 430061,China;Affiliated Hospital of Hubei University of Chinese Medicine,Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430006,China)

机构地区：[1]湖北中医药大学,湖北武汉430061 [2]针灸治未病湖北省协同创新中心,湖北武汉430061 [3]湖北时珍实验室,湖北武汉430061 [4]湖北中医药大学附属医院湖北省中医院,湖北武汉430006

出　　处：《针灸临床杂志》2025年第4期80-87,共8页Journal of Clinical Acupuncture and Moxibustion

基　　金：国家中医药管理局青年岐黄学者培养项目,编号: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,编号:国中医药人教函[2022]75号。

摘　　要：目的:观察电针预处理对心肌缺血再灌注损伤(MIRI)大鼠心肌细胞铁死亡的影响,并探讨其可能的作用机制。方法:45只Sprague-Dawley(SD)大鼠随机分为假手术组(Sham)、模型组(MIRI)和模型+电针预处理组(MIRI+EA),各15只。适应性喂养1周后,MIRI+EA组取双侧“内关”(PC 6)、双侧“足三里”(ST 36)和“关元”(SN 4)进行电针干预20 min,1次/d,连续治疗7 d,其余大鼠捆绑20 min/次,1次/d,连续捆绑7 d,第8天所有大鼠均进行MIRI造模,其中Sham组缝线穿过左冠状动脉前降支下方,但不结扎。造模结束后,TTC染色法检测各组大鼠心肌梗死面积;HE染色法观察心肌细胞病理变化;透射电镜观察线粒体超微结构变化;试剂盒检测大鼠缺血区心肌细胞亚铁(Fe^(2+))、还原型谷胱甘肽(GSH)和氧化谷胱甘肽二硫化物(GSSG)水平;流式细胞仪检测缺血区心肌组织ROS水平;Western blot法检测溶质载体家族7成员11(SLC7A11)、谷胱甘肽过氧化酶4(GPX4)、转铁蛋白受体1(TFR1)、铁蛋白重链1(FTH1)和长链脂酰CoA合成酶4(ACSL4)蛋白表达;免疫组织化学染色法(IHC)检测缺血区心肌组织SLC7A11和GPX4蛋白表达。结果:HE染色及透射电镜结果显示,MIRI组心肌纤维断裂、细胞肿胀变大、线粒体嵴断裂和线粒体膜破碎,MIRI+EA组病理改变明显减轻;与Sham组比较,MIRI组心肌梗死面积显著增加,差异具有统计学意义(P<0.01),ROS和Fe^(2+)水平均明显升高,差异具有统计学意义(P<0.01),GSH/GSSG水平明显降低,差异具有统计学意义(P<0.01),TFR1和ACSL4蛋白表达显著上调,差异具有统计学意义(P<0.01),FTH1、SLC7A11和GPX4蛋白表达均显著下调,差异具有统计学意义(P<0.01),心肌细胞质内SLC7A11和GPX4蛋白阳性表达均显著减少,差异具有统计学意义(P<0.01);与MIRI组比较,MIRI+EA组心肌梗死面积显著减小,差异具有统计学意义(P<0.01),ROS、Fe^(2+)水平均降低,差异具有统计学意义(P<0.01),GSH/GSSG水平明显升高,�Objective:To observe the effect of electro-acupuncture(EA) pretreatment on ferroptosis of myocardial cells in rats with myocardial ischemia-reperfusion injury(MIRI) and to explore its possible mechanism.Methods:A total of 45 SD rats were randomly divided into the Sham operation(Sham) group,the model(MIRI) group and the model plus EA pretreatment group(MIRI+EA),with 15 rats in each group.After 1 week of adaptive feeding,bilateral Neiguan(PC6),Zusanli(ST36) and Guanyuan(RN4) were subjected to EA intervention for 20 min in the MIRI+EA group,once a day for 7 consecutive days,and the rats in the other groups were bound for 20 min without other intervention,once a day for 7 consecutive days.MIRI rat model was established in all rats on the 8th day.In the Sham group,sutures passed below the anterior descending branch of the left coronary artery without ligation.After modeling,the myocardial infarction size was detected by TTC staining,the pathological changes of cardiomyocytes were observed by HE staining,the mitochondrial ultrastructure changes were observed by transmission electron microscopy,and the levels of ferrous(Fe^(2+)),reduced glutathione(GSH) and oxidized glutathione disulfide(GSSG) were detected by kits.Flow cytometry was used to detect ROS level in ischemic myocardium.Western blot assay was used to detect the expressions of 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),transferrin receptor 1(TFR1),ferritin heavy chain 1(FTH1) and long chain fatty acyl CoA synthetase 4(ACSL4).Immunohistochemical staining(IHC) was used to detect the protein expressions of SLC7A11 and GPX4 in ischemic myocardial tissues.Results:The results of HE staining and transmission electron microscopy showed that myocardial fiber breakage,cell swelling,mitochondrial ridge breakage and mitochondrial membrane breakage in the MIRI group,and the pathological changes in the MIRI+EA group were significantly reduced.Compared with those in the Sham group,the myocardial infarction area was significantly increased(P<0.01),the levels of ROS a

关 键 词：心肌缺血再灌注损伤 电针预处理 铁死亡 SLC7A11/GPX4 

分 类 号：R246.1[医药卫生—针灸推拿学]