NLRP3炎性小体在治疗性浅低温后处理大鼠心肌缺血-再灌注中的作用  

Role of NLRP3 inflammasome in therapeutic mild hypothermia post-treated myocardial ischemia-reperfusion rats

作　　者：李亚琦[1] 陈祎轩 张静[2] 余树春[2] LI Yaqi;CHEN Yixuan;ZHANG Jing;YU Shuchun(Department of Anesthesiology,Jiangxi Provincial People's Hospital and the First Afflicated Hospital of Nanchang Medical College,Nanchang 330006,China)

机构地区：[1]江西省人民医院(南昌医学院第一附属医院)麻醉科,330006 [2]南昌大学第二附属医院麻醉科

出　　处：《临床麻醉学杂志》2024年第2期178-184,共7页Journal of Clinical Anesthesiology

基　　金：国家自然科学基金(81760338);江西省中医药管理局科技计划项目(2022B1038);江西省卫生健康委员会科技计划项目(202310106)。

摘　　要：目的 分析NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎性小体在治疗性浅低温(34℃)后处理的大鼠离体心肌缺血-再灌注模型中的作用并探讨其机制。方法 选择清洁级成年雄性SD大鼠60只,7~10周龄,体重250~300 g。采用随机数字表法将大鼠分为五组:空白对照组(S组)、心肌缺血-再灌注组(IR组)、34℃浅低温后处理心肌缺血-再灌注组(MH组)、34℃浅低温后处理心肌缺血-再灌注+3-TYP组(HT组)和34℃浅低温后处理心肌缺血-再灌注+3-TYP+MCC950组(HTM组),每组12只。S组在37℃灌流液平衡灌流大鼠心脏180 min;IR组在37℃灌流液平衡灌流大鼠心脏30 min后,缺血30 min, 37℃灌注液再灌注120 min;MH组在37℃灌流液平衡灌流大鼠心脏30 min后,缺血30 min, 34℃灌注液再灌注120 min;HT组在37℃灌流液平衡灌流大鼠心脏30 min后,缺血30 min,在灌注液中加入沉默信息调节因子2同源蛋白3(sirt3)抑制剂3-TYP后行34℃灌注液再灌注120 min;HTM组在37℃灌流液平衡灌流大鼠心脏30 min后,缺血30 min,在灌注液中加入sirt3抑制剂3-TYP和NLRP3抑制剂MCC950后行34℃灌注液再灌注120 min。再灌注120 min后取离体心脏,采用ELISA法测定灌注后心脏漏液中IL-1β、IL-6浓度,Western blot法检测心肌组织中NLRP3和sirt3蛋白相对含量,1%氯化三苯基四氮唑染色计算心肌梗死面积,HE染色观察心肌病理变化。结果 与S组比较,IR组、MH组、HT组和HTM组再灌注30、60、90、120 min时HR明显减慢,LVSP、dp/dt_(max)明显降低,LVEDP明显升高;心脏漏液中IL-6和IL-1β浓度、心肌梗死面积百分比明显升高(P<0.05);IR组、HT组和HTM组心肌组织中sirt3蛋白含量明显降低,NLRP3蛋白含量明显升高(P<0.05);MH组心肌组织中sirt3和NLRP3蛋白含量明显升高(P<0.05)。与IR组比较,MH组和HTM组再灌注30、60、90、120 min时HR明显增快,LVSP、±dp/dt_(max)明显升高,LVEDP明显降低;心脏漏液中IL-6和IL-1β浓度、心肌梗死面积百分比Objective To analyze the role of NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome in a therapeutic mild hypothermia(34℃)treated isolated rat myocardial ischemia-reperfusion model and explore its mechanism.Methods Sixty clean grade adult male SD rats,aged 7-10 weeks,weighing 250-300 g.Using a random number table method,the rats were divided into five groups:blank control group(group S),myocardial ischemia-reperfusion group(group IR),34℃mild hypothermia post-treated myocardial ischemia-reperfusion group(group MH),34℃mild hypothermia post-treated myocardial ischemia-reperfusion+3-TYP group(group HT),and 34℃mild hypothermia post-treated myocardial ischemia-reperfusion+3-TYP+MCC950 group(group HTM),12 rats in each group.Group S perfused the rat heart at 37℃with a balanced perfusion solution for 180 minutes.Group IR received balanced perfusion of the rat heart at 37℃for 30 minutes,followed by ischemia for 30 minutes and reperfusion with 37℃perfusion for 120 minutes.Group MH perfused the rat heart at 37℃for 30 minutes,followed by ischemia for 30 minutes and reperfusion with 34℃perfusion solution for 120 minutes.Group HT perfused the hearts of rats at 37℃for 30 minutes,followed by ischemia for 30 minutes,silent mating type information regulation 2 homolog 3(sirt3)inhibitor 3-TYP was added to the perfusate,and then perfused at 34℃for 120 minutes.Group HTM perfused the hearts of rats at 37℃for 30 minutes,followed by ischemia for 30 minutes,sirt3 inhibitor 3-TYP and NLRP3 inhibitor MCC950 were added to the perfusate,and then perfused at 34℃for 120 minutes.The isolated heart was obtained 120 minutes after reperfusion,and the concentrations of IL-6 and IL-1βin the perfused cardiac fluid was measured using ELISA method,Western blot method for detecting the relative content of NLRP3 and sirt3 proteins in myocardial tissue,1%triphenyl tetrazolium chloride staining for calculating myocardial infarction area,and HE staining for observing myocardial pathological changes.Res

关 键 词：心肌 缺血-再灌注 治疗性浅低温 NOD样受体热蛋白结构域相关蛋白3 沉默信息调节因子2同源蛋白3 炎性小体 线粒体 

分 类 号：R614[医药卫生—麻醉学]