沉默IL36R通过SIRT1/FOXO1/DRP1介导心肌细胞线粒体分裂减轻体外循环机心肌缺血再灌注损伤  

Silencing IL36R alleviates myocardial ischemia-reperfusion injury via SIRT1/FOXO1/DRP1 mediated mitochondrial fission of cardiomyocytes

作　　者：陈基鸿 魏永钧 孟子龙 郑宝石[1] Chen Jihong;Wei Yongjun;Meng Zilong;Zheng Baoshi(Department of Cardiothoracic Surgery,the First Affiliated Hospital of Guangxi Medical University,Nanning 530000,China;不详)

机构地区：[1]广西医科大学第一附属医院心胸外科,南宁530000

出　　处：《中国循证心血管医学杂志》2025年第1期22-27,共6页Chinese Journal of Evidence-Based Cardiovascular Medicine

基　　金：国家自然科学基金(8206020365)。

摘　　要：目的通过建立深低温停循环(DHCA)模型,探讨抑制白介素36受体(IL36R)对深低温停循环(DHCA)大鼠心肌细胞线粒体的影响及相关机制。方法42只雄性成年SPF级的SD大鼠,随机分为七组(每组各6只):SHAM组(对照组)、DHCA组(深低温停循环组)、Si-IL36R+DHCA组(转染沉默病毒+深低温停循环)、Si-NC+DHCA组(转染沉默空载病毒+深低温停循环)、OE-IL36R+DHCA组(转染过表达病毒+深低温停循环)、OE-NC+DHCA组(转染过表达空载病毒+深低温停循环)、NAM+DHCA组(抑制剂+深低温停循环)。病毒转染组术前1个月,通过大鼠尾静脉注射IL36R过表达或者沉默病毒,同时注射相对应的空载病毒,NAM+DHCA组术前1 h注射沉默信息调节因子1(SIRT1)抑制剂尼克酰胺(NAM)。除SHAM组外,其余六组建立体外循环(CPB)模型,经历降温、停循环、复温过程,全过程2 h。苏木精-伊红(HE)染色观察心脏组织形态变化,透射电镜观察心肌细胞线粒体形态,血清检测肌酸激酶同工酶MB(CKMB)、乳酸脱氢酶(LDH)水平,心肌组织匀浆检测超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)水平,Western Blot检测通路蛋白SIRT1、FOXO1,线粒体动力蛋白DRP1及其磷酸化蛋白p-DRP1(Ser616)、p-DRP1(Ser637)表达水平。结果沉默IL36R后减轻了大鼠体外循环后心肌组织损伤以及线粒体损伤,同时降低了心肌氧化应激水平,SOD、GSH活性上升、MDA含量下降(P<0.05),SIRT1、FOXO1、p-DRP1(Ser637)的蛋白表达上升(P<0.05),抑制了p-DRP1(Ser616)蛋白表达(P<0.05),反之,过表达IL36R或抑制SIRT1后,心肌组织损伤以及线粒体损伤加重,氧化应激水平升高,SIRT1、FOXO1、p-DRP1(Ser637)的蛋白表达下降(P<0.05),促进p-DRP1(Ser616)蛋白表达(P<0.05)。结论沉默IL36R在体外循环后心肌缺血再灌注损伤中起心肌保护作用,其保护机制可能是通过上调SIRT1/FOXO1抑制DRP1介导的心肌细胞线粒体过度分裂实现的。Objective By establishing a deep hypothermic circulatory arrest(DHCA)model,this study explores the effects of inhibiting interleukin-36 receptor(IL36R)on the mitochondria of cardiomyocytes in rats undergoing DHCA and the related mechanisms.Methods Forty-two male adult SPF SD rats were randomly divided into 7 groups(6 rats per group):SHAM group(control group),DHCA group(cryopreservation group),Si-IL36R+DHCA group(transfection of silenced virus+cryopreservation group),Si-NC+DHCA group(transfection of silenced no-load virus+cryopreservation group),OE-IL36R+DHCA group(transfection of overexpressed virus+cryopreservation group),OE-NC+DHCA group(overexpression of no-load virus+cryopreservation),NAM+DHCA group(inhibitor+cryopreservation).In the viral transfection group,IL36R overexpressed or silenced virus was injected through the rat tail vein one month before surgery,and the corresponding no-load virus was injected simultaneously.In the NAM+DHCA group,the silencing information regulatory factor 1(SIRT1)inhibitor Nicotinamide(NAM)was injected 1 h before surgery.In addition to SHAM group,the other 6 were used to construct stereoscopic external circulation(CPB)model,and underwent cooling,stopping circulation,and rewarming for 2 hours.The morphological changes of cardiac tissue were observed by hematoxylin-eosin(HE)staining,mitochondrial morphology of cardiomyocytes was observed by transmission electron microscopy,the levels of creatine kinase isoenzyme MB(CKMB)and lactate dehydrogenase(LDH)in serum,and the levels of superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione(GSH)in myocardial tissue homogenate were detected.Western Blot analysis was performed to detect the expression levels of pathway proteins SIRT1,FOXO1,mitochondrial dynamin DRP1 and its phosphorylated proteins p-DRP1(Ser616)and p-DRP1(Ser637).Results Silenced IL36R alleviated myocardial tissue damage and mitochondrial damage after cardiopulmonary bypass in rats,decreased myocardial oxidative stress level,increased SOD and GSH activities,decreased MD

关 键 词：体外循环 缺血再灌注损伤 IL36R 线粒体损伤 

分 类 号：R654.1[医药卫生—外科学]