复氧前给予不同浓度乳化异氟醚的大鼠缺氧/复氧心肌细胞损伤情况观察  

Observation of hypoxic/reoxygenated cardiomyocyte injury in rats given different concentrations of emulsified isoflurane before reoxygenation

作　　者：王佳琪 徐鹏 陈伟 李晓娟 王海英 喻田 WANG Jiaqi;XU Peng;CHEN Wei;LI Xiaojuan;WANG Haiying;YU Tian(The Affiliated Hospital of Zunyi Medical University,Zunyi 563000,China;不详)

机构地区：[1]遵义医科大学附属医院,贵州遵义563000 [2]湘潭市中心医院

出　　处：《山东医药》2020年第9期36-39,共4页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(30960366)。

摘　　要：目的观察复氧前给予不同浓度乳化异氟醚(EI)的大鼠缺氧/复氧心肌细胞损伤情况。方法手术分离雄性SD大鼠心肌细胞并培养,将心肌细胞随机分为FAT组和不同浓度EI处理组(EI1组、EI2组、EI3组),采用混合气体培养法制备缺氧/复氧心肌细胞,其中FAT组复氧前加入脂肪乳(FAT)进行处理,EI1组、EI2组、EI3组复氧前分别加入0.84、1.68、2.52 mmol/L EI进行处理。收集各组心肌细胞,进行细胞线粒体损伤评分(Flameng评分),测定细胞内游离Ca2+荧光强度,观察细胞内NF-E2相关因子2(Nrf2)蛋白核转位荧光强度,采用RT-PCR法和Western blotting法分别检测各组细胞中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和蛋白。结果不同浓度EI处理组细胞线粒体Flameng评分和细胞内游离Ca2+荧光强度均低于FAT组,且EI2组最低。EI1组、EI2组、EI3组、FAT组细胞内Nrf2蛋白核转位荧光强度分别为152.71±13.36、181.95±10.59、156.39±11.20、135.25±19.73,其中EI2组细胞内Nrf2核转位荧光强度与EI1组、EI3组、FAT组相比,P均<0.05。不同浓度EI处理组Nrf2、HO-1、SOD1、NQO1 mRNA和蛋白相对表达量均高于FAT组,且EI2组最高。结论复氧前给予不同浓度EI处理对大鼠缺氧/复氧心肌细胞均可起到保护作用,1.68 mmol/L的EI保护作用最优;EI可能是通过激活Nrf2/ARE通路发挥保护作用。Objective To observe the hypoxia/reoxygenated cardiomyocyte injury in rats given different concentrations of emulsified isoflurane(EI)before reoxygenation.Methods We isolated and cultured the cardiomyocytes from male SD rats,and divided the cardiomyocytes into the group FAT and group EI(treat with different concentrations of emulsified isoflurane:groups EI1,EI2,and EI3).The cardiomyocytes were exposed to the mixed air to establish the cardiomyocyte H/R damage models;during this process,the cardiomyocytes in the group FAT were treated with fat emulsion(FAT)before reoxygenation,cells in the group EI1,group EI2,and group EI3 were treated with 0.84,1.68,and 2.52 mmol/L EI before reoxygenation,respectively.We collected cardiomyocytes from each group,performed cell mitochondrial injury score(Flameng score),measured the intracellular free Ca2+fluorescence intensity,and observed the nuclear translocation fluorescence intensity of NF-E2 related factor 2(Nrf2)protein in cells.The expression levels of Nrf2,heme oxygenase-1(HO-1),superoxide dismutase 1(SOD1),quinine oxidoreductase 1(NQO1)protein and mRNA were detected by using real-time polymerase chain reaction and Western blotting.Results The mitochondrial Flameng score and intracellular free Ca2+fluorescence intensity of cells in the group EI were lower than those of group FAT,and group EI2 was the lowest.The Nrf2 protein nuclear translocation fluorescence intensities in the group EI1,group EI2,group EI3,and group FAT were 152.71±13.36,181.95±10.59,156.39±11.20,and 135.25±19.73,respectively,with statistically significant difference between the group EI2 and the group E11,group EI3 and group FAT(all P<0.05).The expression levels of Nrf2,HO-1,SOD1,NQO1 mRNA and protein in the group EI were higher than those in the FAT group,with the highest level in the group EI2.Conclusion Treatment with EI at different concentrations before reoxygenation can protect cardiomyocytes from hypoxic/reoxygenated injury,and the protective effect of EI at 1.68 mmol/L is the best;EI may play a

关 键 词：乳化异氟醚 心肌细胞 缺氧/复氧损伤 NF-E2相关因子2/抗氧化反应原件通路 心肌保护 

分 类 号：R310.17[医药卫生—基础医学]