ROS在二氮嗪后处理上调HIF-1α表达减轻大鼠离体心脏缺血再灌注损伤中的作用  

Role of ROS in diazoxide postconditioning-induced upregulation of hypoxia inducible factor-1 to alleviate myocardial ischemia-reperfusion injury in isolated rat hearts

作　　者：周雯静 李进 张琳 喻田 王海英 Zhou Wenjing;Li Jin;Zhang Lin;Yu Tian;Wang Haiying(Guizhou Key Laboratory of Anesthesia and Organ Protection,Affiliated Hospital of Zunyi Medical University,Zunyi 563000,China;Department of Anesthesiology, Affiliated Hospital of Southwest Medical University, Luzhou 646003,China;Department of Anesthesiology,the Affiliated Hospital of Zunyi Medical University,Zunyi 563000,China)

机构地区：[1]遵义医科大学附属医院,贵州省麻醉与器官保护基础研究重点实验室,563000 [2]西南医科大学附属医院麻醉科,泸州646003 [3]遵义医科大学附属医院麻醉科,563000

出　　处：《中华麻醉学杂志》2020年第12期1520-1524,共5页Chinese Journal of Anesthesiology

摘　　要：目的评价活性氧(ROS)在二氮嗪后处理上调缺氧诱导因子-1α(HIF-1α)表达减轻大鼠离体心脏缺血再灌注损伤中的作用。方法SPF级健康成年雄性SD大鼠,体重250~280 g。制备成功的离体心脏模型72个,采用随机数字表法分为4组(n=18):对照组(C组)、心脏缺血再灌注组(I/R组)、二氮嗪后处理组(DPO组)和ROS清除剂N-(2-巯基丙酰)-甘氨酸+二氮嗪后处理组(MPO组)。C组采用K-H液持续灌注120 min;I/R组采用K-H液灌注20 min,随后灌注4℃的ST.Thomas心脏停跳液40 min,再灌注K-H液60 min;DPO组于再灌注即刻灌注含二氮嗪50μmol/L的K-H液5 min,继续灌注K-H液55 min;MPO组于再灌注即刻灌注含MPG 2 mmol/L的K-H液3 min,随后灌注含二氮嗪的K-H液5 min,继续灌注K-H液52 min。于平衡灌注20 min和再灌注120 min时分别记录HR、LVEDP、LVDP和+dp/dtmax。于灌注120 min时取心肌组织,TTC法检测左心室心肌梗死体积;电镜下观察心肌超微结构,行线粒体Flameng评分;采用RT-PCR和Western blot法分别检测HIF-1α、诱导型一氧化氮合酶(iNOS)、血红素加氧酶-1(HO-1)及血管内皮生长因子(VEGF)及其mRNA的表达。结果与C组比较,I/R组HR、LVDP、+dp/dtmax降低,LVEDP、心肌梗死体积百分比和线粒体Flameng评分增加,VEGF、HO-1、iNOS及其mRNA和HIF-1α表达上调(P<0.05);与I/R组比较,DPO组HR、LVDP和+dp/dtmax升高,LVEDP、心肌梗死体积百分比和线粒体Flameng评分减少,VEGF、HO-1和iNOS及其mRNA和HIF-1α表达上调(P<0.05);与DPO组比较,MPO组HR、LVDP和+dp/dtmax降低,LVEDP、心肌梗死体积百分比和线粒体Flameng评分增加,VEGF、HO-1、iNOS及其mRNA和HIF-1α表达下调(P<0.05)。结论ROS参与了二氮嗪后处理上调HIF-1α表达减轻大鼠离体心脏缺血再灌注损伤的过程。Objective To evaluate the role of reactive oxygen species(ROS)in diazoxide postconditioning-induced upregulation of hypoxia inducible factor-1(HIF-1α)to alleviate myocardial ischemia-reperfusion(I/R)injury in isolated rat hearts.Methods SPF healthy adult male Sprague-Dawley rats,weighing 250-280 g,were used in this study.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5%CO2.Seventy-two isolated rat hearts with I/R injury were divided into 4 groups(n=18 each)using a random number table method:control group(C group),I/R group,diazoxide postconditioning group(DPO group),and ROS scavenger N-(2-mercaptopropionyl)-glycine plus diazoxide postconditioning group(MPO group).Hearts were continuously perfused with K-H solution for 120 min in group C.In group I/R,the hearts were perfused with K-H solution for 20 min,followed by ST.Thomas cardioplegia solution at 4℃for 40 min,and then with K-H solution for 60 min.In DPO group,the hearts were perfused with K-H solution containing 50μmol/L diazoxide for 5 min starting from the time point immediately after onset of reperfusion,and then with K-H solution for 55 min.In MPO group,the hearts were perfused with K-H solution containing MPG 2 mmol/L for 3 min starting from the time point immediately after onset of reperfusion,followed by perfusion with K-H solution containing diazoxide for 5 min,and then with K-H solution for 52 min.Heart rate(HR),left ventricular end-diastolic pressure(LVEDP),left ventricular developed pressure(LVDP),and+dp/dtmax were recorded at 20 min of equilibration and 120 min of reperfusion.The myocardial tissues were removed at the end of reperfusion to measure the myocardial infarct size of left ventricle and observe the ultrastructure.The Flameng score of mitochondria was performed.The expression of hypoxia inducible factor-1α(HIF-1α),inducible nitric oxide synthase(iNOS),heme oxygenase-1(HO-1)and vascular endothelial growth factor(VEGF)protein and mRNA in myocardial tissues was detected by real-ti

关 键 词：活性氧 二氮嗪 缺血后处理 缺氧诱导因子1 Α亚基 心肌再灌注损伤 

分 类 号：R363[医药卫生—病理学]