基础医学与临床 ›› 2019, Vol. 39 ›› Issue (4): 552-557.

• 研究论文 • 上一篇    下一篇

丙泊酚减轻过氧化氢诱导的心肌细胞系H9C2损伤

王磊,朱自强,袁丽霞,袁孝伟   

  1. 郑州市第七人民医院
  • 收稿日期:2018-09-25 修回日期:2019-01-24 出版日期:2019-04-05 发布日期:2019-03-26
  • 通讯作者: 王磊 E-mail:2672578697@qq.com

Propofol alleviates myocardial injury induced by hydrogen peroxide in cell line H9C2

  • Received:2018-09-25 Revised:2019-01-24 Online:2019-04-05 Published:2019-03-26

摘要: 目的 研究丙泊酚对过氧化氢(H2O2)诱导的心肌细胞系H9C2损伤及活性氧(ROS)水平的影响。方法 将心肌细胞分成对照组、H2O2处理组和15、30和60 μmol/L丙泊酚干预组。MTT检测细胞活力;二硝基苯肼显色法检测乳酸脱氢酶(LDH)漏出率;流式细胞计量术检测细胞凋亡;Western blot检测细胞中激活型caspase-3(cleaved caspase-3)、胞浆和线粒体中细胞色素C(Cytochrome C)蛋白水平;黄嘌呤氧化法检测超氧化物歧化酶 ( SOD)活性;钼酸铵法检测过氧化氢酶(CAT)活性;二硫代二硝基苯甲酸法检测谷胱甘肽过氧化物酶(GSH-Px)活性;硫代巴比妥酸法检测丙二醛(MDA)含量;二氯二氢荧光素-乙酰乙酸酯法检测ROS活性;JC-1法检测线粒体膜电位。结果 与对照组比较,H2O2处理组细胞活力降低;LDH漏出率和凋亡率升高;cleaved caspase-3蛋白水平及胞浆中Cytochrome C水平升高;线粒体Cytochrome C水平降低;SOD、CAT和GSH-Px活性降低;MDA及ROS含量升高;线粒体膜电位下降(P<0.05)。与H2O2处理组比较,15、30和60 μmol/L丙泊酚组细胞活性升高;LDH漏出率降低;凋亡率和cleaved caspase-3水平降低;胞质中Cytochrome C蛋白水平减少;线粒体中Cytochrome C水平升高;细胞SOD活性、CAT活性和GSH-Px活性升高;MDA含量和ROS含量降低;线粒体线粒体膜电位升高(P<0.05)。结论 丙泊酚可以减轻H2O2诱导的心肌细胞损伤,减少细胞凋亡并降低细胞ROS水平。

关键词: 心肌细胞, 过氧化氢, ROS, 凋亡, 丙泊酚

Abstract: Objective To study the effects of propofol on myocardial injury and ROS level induced by H2O2. Methods Cardiac myocytes are treated with H2O2 and propofol,as control group, H2O2 group and 15, 30 and 60 μmol/L propofol group.MTT detection of cell viability, the leakage rate of LDH was detected by two nitrophenyl hydrazine colorimetric assay, cell apoptosis was detected by flow cytometry, Western blot was used to detect the level of cleaved caspase-3 protein in cells and the level of Cytochrome C protein in cytoplasm and mitochondria, SOD activity was detected by xanthine oxidation, CAT activity was detected by ammonium molybdate method, the activity of GSH-Px was detected by two thio two Nitrobenzoic Acid method, thiobarbituric acid method was used to detect the content of MDA, the level of ROS was detected by two chloro two fluorescein acetoacetate method, the JC-1 method was used to detect the mitochondrial membrane potential. Results Compared with control group,the activity of myocardial cells decreased in H2O2 group, the leakage rate of LDH was increased, the rate of apoptosis increased, the level of cleaved caspase-3 protein in the cells increased, SOD activity, CAT activity and GSH-Px activity reduced, the content of MDA in the cells increased and the content of ROS increased, the membrane potential of cell mitochondria decreased, the level of Cytochrome C protein in the cytoplasm increased, the levels of Cytochrome C protein in mitochondria decreased(P<0.05). Compared with H2O2 group, the activity of cardiomyocytes in propofol group improved, the leakage rate of LDH decrease , the rate of apoptosis and the level of cleaved caspase-3 protein in cells decreased, SOD activity , CAT activity and GSH-Px activity in cells increased, the content of MDA and ROS in cells decreased, mitochondrial mitochondrial membrane potential improved, the Cytochrome C protein level in the cytoplasm reduced, the level of Cytochrome C protein in mitochondria improved(P<0.05). Conclusions Propofol alleviated the injury of cardiomyocytes induced by H2O2, reduce cell apoptosis and reduce the level of ROS in cells.

Key words: cardiomyocytes, hydrogen peroxide, ROS, apoptosis, Propofol