依达拉奉对阿霉素诱导的心肌细胞毒性的影响及其机制

Effect of Edaravone on Adriamycin-induced Myocardial Toxicity and Its Possible Mechanisms

目的观察依达拉奉对蒽环类抗癌药阿霉素诱导的乳鼠心肌细胞毒性的影响及其机制。方法用不同浓度的阿霉素处理乳鼠心肌细胞,建立蒽环类抗癌药心脏毒性的体外模型。依达拉奉在阿霉素处理心肌细胞前1 h加入培养液中作为预处理。应用CCK-8比色法检测细胞存活率;谷胱甘肽试剂盒检测细胞内还原型谷胱甘肽的水平;双氯荧光素染色/荧光显微镜照相检测细胞内活性氧的含量;Western blot法检测胞浆中细胞色素C和Cleaved Caspase-3的表达。结果阿霉素在1-8 mg/L浓度范围内处理乳鼠心肌细胞24 h,可剂量依赖性地降低细胞存活率。在2 mg/L阿霉素处理心肌细胞前1 h,应用不同浓度的依达拉奉预处理可明显地抑制阿霉素诱导的心肌细胞毒性损伤,使细胞存活率显著升高。20μmol/L依达拉奉预处理1 h可抑制2 mg/L阿霉素引起的氧化应激反应,使胞内还原型谷胱甘肽水平升高,活性氧含量降低。在阿霉素损伤心肌细胞前,给予依达拉奉预处理也可降低胞浆中Cytochrome C含量及Cleaved Caspase-3表达。结论依达拉奉可显著减弱阿霉素诱导的心肌细胞毒性,此保护作用可能与抗氧化及抑制凋亡相关蛋白的表达有关。

Aim To explore the effect of edaravone（EDA） on myocardial toxicity –induced by anthracycline antitumor adriamycin（ADR） and the mechanisms underlying. Methods Primary cultured myocardial cells were treated with ADR at different concentrations as a cardiac toxicity model of anthracycline antitumor.EDA was administrated 1 h before ADR as pretreatment.Cell viability was measured by using cell counter kit（CCK-8）.The level of intercellular reduced glutathinone（GSH） was detected according to commercial kit.Intercellular reactive oxygen species（ROS） was observed by DCFH-DA staining and photofluorography.The expressions of Cytochrome C and cleaved Caspase-3 were detected by western blot. Results ADR at the concentrations from 1 to 8 mg/L for 24 h damaged myocardial cells in a dose-dependent manner.Preconditioning of 5-20 μmol/L EDA protected myocardial cells against ADR-induced injury,increasing cell survival rate.The preconditioning of EDA inhibited oxidative stress induced by 2 mg/L ADR,increasing the level of GSH and decreasing the content of ROS,and attenuated the expressions of Cytochrome C and cleaved Caspase-3. Conclusion EDA can attenuate the myocardial toxicity induced by ADR,which may be associated with its anti-oxidation and anti-apoptosis action.