白藜芦醇通过上调沉默信息调节因子1调节香烟提取物诱导心肌线粒体产生活性氧簇的机制

SIRT1 regulates cigarette smoke-induced production of reactive oxygen species in myocardial mitochondria: Role of resveratrol

目的探讨沉默信息调节因子1(SIRT1)调节香烟提取物(CSE)诱导心肌线粒体产生活性氧簇(ROS)的机制及白藜芦醇(Res)心肌保护作用。方法 H9C2细胞分5组:C组(对照组)、二甲亚砜组(DMSO溶剂对照组)、CSE组(CSE刺激组)、CSE+Res组、Res组。采用Taqman探针荧光定量PCR检测各组SIRTI的mRNA表达、Weston Blot检测各组细胞SIRTI蛋白表达、荧光分光光度计检测细胞线粒体膜电位、荧光微量平板法检测不同浓度CSE刺激H9C2细胞产生ROS的变化、特异性荧光探针CM-H2DCFDA检测活细胞内ROS变化。结果与C组相比,CSE组SIRT1mRNA及蛋白质表达显著降低,ROS表达显著增强。与CSE组相比,CSE+Res组SIRT1mRNA及蛋白质表达升高,ROS表达显著减少。随CSE刺激浓度增加,线粒体膜电位降低,ROS产生增加。提前给予Res可使线粒体膜电位升高,ROS产生减少。结论 Res可通过刺激SIRT1的表达而抑制CSE诱导的心肌细胞线粒体功能受损、ROS的释放,这可能是其吸烟相关慢性阻塞性肺疾病诱导的心脏氧化应激损伤中发挥心脏保护作用的机制之一。

Objective We aimed to explore the regulation of SIRT1 in cigarette smoke-induced reactive oxygen species（ROS） release in myocardial mitochondria,and the protective effect of resveratrol（Res） in myocardium.Methods In vitro,H9C2 cells were divided into C group（control group）,dimethylsulfoxide group（solvent control group）,CSE group（cigarette smoke extract group）,CSE group with Res treatment group（CSE＋Res group）,and Res group.RT-PCR was employed for the determination of the expression of SIRT1 mRNA.The protein of SIRT expression was observed by Weston Blot.Mitochondrial membrane potential was measured by fluorescence spectrophotometer.Specific florescent probe CM-H2DCFDA and fluorescent micro-plate method was employed for the determination of ROS expression.Results The expression of ROS increased in H9C2 cells of CSE group against the cells of C group（P0.05）,whereas the expression of SIRT1 decreased（P0.05）.Compared to CSE group,the expression of ROS decreased in CSE＋Res group（P0.05）,and the expression of SIRT1increased.The mitochondria membrane potential drop（P0.05） in CSE group,whereas can be increased by Res（P 0.05）.Conclusions The myocardial protection of resveratrol against CSE-induced ROS release.In H9C2 cells,Res,by simulating SIRT expression,attenuates CSE-induced the oxidant stress and mitochondrial dysfunction.The myocardial protection of resveratrol could potentially serve as therapeutic value of oxidative stress injury induced by smoking-related COPD in cardiovascular system.