miR-155-5p通过调控心肌细胞焦亡对大鼠心肌缺血-再灌注损伤的影响及机制研究  

Effect and mechanism of miR-155-5p on myocardial ischemia-reperfusion injury in rats by regulating myocardial pyroptosis

作　　者：卢秋玉[1] 陈燕青 申庆荣[1] 李鑫[3] 夏冰雨 苏金妹 Lu Qiuyu;Chen Yanqing;Shen Qingrong;Li Xin;Xia Bingyu;Su Jinmei(Department of Pharmacy,the People’s Hospital ofGuangxi Zhuang Autonomous Region,Nanning 530021,China;不详)

机构地区：[1]广西壮族自治区人民医院药学部,南宁530021 [2]广西壮族自治区江滨医院药学部 [3]广西中医药大学 [4]广西国际壮医医院明秀分院药剂科

出　　处：《器官移植》2024年第6期903-911,共9页Organ Transplantation

基　　金：广西自然科学基金项目(2023GXNSFBA026095)。

摘　　要：目的探讨微小RNA(miR)-155-5p对心肌缺血-再灌注损伤(IRI)大鼠心肌细胞焦亡的影响及机制。方法60只SD大鼠随机分为假手术(sham)组、IRI组、agomir-NC组、miR-155-5p agomir组、antagomir-NC组和miR-155-5p antagomir组,每组10只。采用超声心动图仪测定大鼠左心室舒张末期内径(LVEDD)、左心室收缩末期内径(LVESD)、左心室射血分数(LVEF)和左心室短轴缩短率(LVFS)。酶联免疫吸附试验检测大鼠血清肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)和心肌肌钙蛋白T(cTnT)水平,心肌组织白细胞介素(IL)-1β、IL-6、IL-18和肿瘤坏死因子(TNF)-α水平;苏木素-伊红染色观察大鼠心肌组织病理学变化;实时荧光定量聚合酶链反应检测大鼠心肌组织miR-155-5p和沉默信息调节因子1(SIRT1)信使RNA(mRNA)表达水平;双荧光素酶报告基因实验验证miR-155-5p与SIRT1的靶向关系;蛋白质印迹法检测大鼠心肌组织SIRT1、NOD样受体蛋白3(NLRP3)、剪切型半胱氨酸天冬氨酸蛋白水解酶-1(Cleaved Caspase-1)和GSDMD蛋白表达水平。结果与sham组比较,IRI组大鼠LVEDD和LVESD升高,LVEF和LVFS降低,血清CK-MB、LDH和cTnT水平升高,心肌组织IL-1β、IL-6、IL-18、TNF-α水平升高,心肌组织结构破坏严重,心肌纤维排列紊乱,NLRP3、Cleaved Caspase-1和GSDMD蛋白相对表达量升高,SIRT1蛋白相对表达量降低(均为P<0.05/5)。与IRI组比较,miR-155-5p agomir组大鼠LVEDD和LVESD升高,LVEF和LVFS降低,血清CK-MB、LDH和cTnT水平升高,心肌组织中IL-1β、IL-6、IL-18、TNF-α水平升高,心肌组织病变程度加重,NLRP3、Cleaved Caspase-1和GSDMD蛋白相对表达量升高,SIRT1蛋白相对表达量降低;miR-155-5pantagomir组大鼠LVEDD和LVESD降低,LVEF和LVFS升高,血清CK-MB、LDH和cTnT水平降低,心肌组织中IL-1β、IL-6、IL-18、TNF-α水平降低,心肌组织病变程度减轻,NLRP3、Cleaved Caspase-1和GSDMD蛋白相对表达量下降,SIRT1蛋白相对表达量升高(均为P<0.05/5)。miR-155-5pObjective To explore the effect and mechanism of microRNA(miR)-155-5p on myocardial pyroptosisin rats with myocardial ischemia-reperfusion injury(IRI).Methods Sixty SD rats were randomly divided into shamgroup,IRI group,agomir-NC group,miR-155-5p agomir group,antagomir-NC group,and miR-155-5p antagomir group,with 10 rats in each group.Echocardiography was used to measure the left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),left ventricular ejection fraction(LVEF),and left ventricular fractionalshortening(LVFS)of rats.Enzyme-linked immune absorbent assay(ELISA)was used to detect the levels of creatinekinase isoenzyme(CK-MB),lactate dehydrogenase(LDH),and cardiac troponin T(cTnT)in serum,as well as the levelsof interleukin(IL)-1β,IL-6,IL-18,and tumor necrosis factor(TNF)-αin myocardial tissue of rats.Hematoxylin-eosinstaining was used to observe pathological changes in rat myocardial tissue.Real-time fluorescent quantitative polymerasechain reaction was used to detect the expression levels of miR-155-5p and silent information regulator 1(SIRT1)messenger RNA(mRNA)in myocardial tissue of rats.Dual-luciferase reporter gene assay was used to verify the targetingrelationship between miR-155-5p and SIRT1.Western blot was used to detect the expression levels of SIRT1,NOD-likereceptor protein 3(NLRP3),cleaved cysteine aspartate specific proteinase-1(Cleaved Caspase-1),and gasdermin D(GSDMD)proteins in myocardial tissue of rats.Results Compared with the sham group,the LVEDD and LVESD of ratsin the IRI group were increased,LVEF and LVFS were decreased,serum levels of CK-MB,LDH,and cTnT wereincreased,IL-1β,IL-6,IL-18 and TNF-αlevels in myocardial tissue were increased,myocardial tissue structure wasseverely damaged,myocardial fibers were disordered,relative expression of NLRP3,Cleaved Caspase-1,and GSDMDproteins were increased,and the relative expression of SIRT1 protein was decreased(all P<0.05/5).Compared with the IRIgroup,the rats in the miR-155-5p agomir group had increased

关 键 词：心肌 缺血-再灌注损伤 微小RNA 沉默信息调节因子1 NOD样受体蛋白3 细胞焦亡 炎症因子 炎症小体 

分 类 号：R617[医药卫生—外科学] R654.2[医药卫生—临床医学]