Nrf2-ARE信号通路在乳化异氟醚后处理减轻大鼠心肌细胞缺氧复氧损伤中的作用  

Role of Nrf2-ARE signaling pathway in mitigation of anoxia/reoxygenation-induced damage to rat cardiomyocytes by emulsified isoflurane postconditioning

作　　者：李小娟 王海英[2] 杜文娟 喻田[2] 

机构地区：[1] 湘潭市中心医院麻醉科 [2]遵义医学院附属医院麻醉科,563000 [3] 山西长治市人民医院正骨科

出　　处：《中华麻醉学杂志》2014年第8期992-995,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（30960366）

摘　　要：目的 评价转录因子NF-E2相关因子2（Nrf2）-抗氧化反应元件（ARE）信号通路在乳化异氟醚后处理减轻大鼠心肌细胞缺氧复氧损伤中的作用.方法 原代培养16 - 20周龄SD大鼠心肌细胞,以104/cm2的细胞密度平铺于层粘连蛋白预处理过的6孔板中孵育20 h.采用随机数字表法,将其分为3组（n=12）：对照组（C组）常规培养110 min;缺氧复氧组（A/R组）心肌细胞缺氧45 min,复氧60 min;乳化异氟醚后处理组（EI组）心肌细胞缺氧45 min时以1.68 mmol/L乳化异氟醚孵育5 min,随后复氧60 min.于复氧末电镜下观察心肌细胞超微结构,并进行线粒体功能评分.采用Real-TimePCR和Western blot法检测心肌细胞Nrf2、血红素加氧酶1（HO-1）、超氧化物歧化酶1（SOD1）、醌氧化还原酶1（NQO1） mRNA及其蛋白表达水平.采用荧光共聚焦显微镜观察Nrf2的核转位情况,记录心肌细胞核内Nrf2活性.结果 与C组比较,A/R组和EI组心肌细胞线粒体损伤评分升高,心肌细胞Nrf2、HO-1、SOD1和NQO1 mRNA及其蛋白表达下调,心肌细胞核内Nrf2活性增强（P＜0.05）;与A/R组比较,EI组心肌细胞线粒体损伤程度评分降低,心肌细胞Nrf2、HO-1、SOD1和NQO1 mRNA及其蛋白表达上调,心肌细胞核内Nrf2活性增强（P＜0.05）.结论 乳化异氟醚后处理减轻大鼠心肌细胞缺氧复氧损伤的机制可能与诱导Nrf2核转位从而激活Nrf2-ARE信号通路有关.Objective To evaluate the role of nuclear factor E2-related factor 2 （Nrf2）-antioxidant response element （ARE） signaling pathway in mitigation of anoxia/reoxygenation （A/R）-induced damage to rat cardiomyocytes by emulsified isoflurane postconditioning.Methods Primarily cultured cardiomyocytes obtained from Sprague-Dawley rats,aged 16-20 weeks,were seeded into the laminin pre-treated 6-well plates at a density of 104/cm2 and cultured for 20 h.The cells were randomly divided into 3 groups （n =12 each） using a random number table：control group （group C）,group A/R,and emulsified isoflurane postconditioning group （group EI）.The cells were cultured for 110 min without any treatment in group C.The cells were exposed to 45 min anoxia followed by 60 min reoxygenation in A/R and EI groups.After 45 min of hypoxia,the cells were cultured with emulsified isoflurane 1.68 mmol/L for 5 min before onset of reoxygenation in group EI.At the end of reoxygenation,transmission electron microscope was used to observe the ultrastructure of myocardial cells,and mitochondrial function was scored.The mRNA and protein expression of Nrf2,heme oxygenase-1 （HO-1）,superoxide dismutase 1 （SOD1）,and quinone oxidoreductase 1 （NQO1） was detected by real-time PCR and Western blot.Laser scanning confocal microscopy was used to detect Nrf2 nuclear translocation at the end of incubation,and Nrf2 activity was recorded in the nucleus of myocardial cells.Results Compared with group C,the mitochondrial function score was significantly increased,the mRNA and protein expression of Nrf2,HO-1,SOD1 and NQO1 was down-regulated,and Nrf2 activity in the nucleus was enhanced in A/R and EI groups.Compared with group A/R,the mitochondrial function score was significantly decreased,the mRNA and protein expression of Nrf2,HO-1,SOD1 and NQO1 was up-regulated,and Nrf2 activity in the nucleus was enhanced in group EI.Conclusion The mechanism by which emulsified isoflurane postconditioning mitigates A/R-induced damage to rat cardiomyoc

关 键 词：NF-E2相关因子2 心肌再灌注损伤 异氟醚 脂肪乳剂 静脉注射用 

分 类 号：R614[医药卫生—麻醉学]