ERK1/2介导依达拉奉保护H9c2心肌细胞对抗异丙肾上腺素诱导的损伤作用  

ERK1/2 mediates edaravone-triggered protection against myocardial damage induced by isoprenaline in H9c2 cells

作　　者：黄涌[1] 王秀玉[2] 傅璐[2] 杨春涛[2] 莫利球[3] 杨战利[2] 董小变[4,5] 廖新学[4,5] 冯鉴强[2,3] 

机构地区：[1]中山大学附属第一医院东山院区内科,广东广州510080 [2]中山大学附属第一医院中山医学院生理教研 ,广东广州510080 [3]中山大学附属第一医院附属第一医院黄埔院区麻醉科 ,广东广州510080 [4]中山大学附属第一医院附属第一医院心血管内科 ,广东广州510080 [5]中山大学附属第一医院高血压血管病科 ,广东广州510080

出　　处：《南方医科大学学报》2010年第12期2663-2666,共4页Journal of Southern Medical University

基　　金：广东省科技计划项目(2009B080701014;2007B080701030)

摘　　要：目的探讨胞外信号调节激酶1/2(ERK1/2)在依达拉奉(EDA)保护H9c2心肌细胞对抗异丙肾上腺素(ISO)损伤中的作用。方法用不同浓度的ISO处理H9c2心肌细胞,建立β1肾上腺素受体持续兴奋诱导心脏毒性的体外模型。EDA在ISO处理心肌细胞前1h加入培养基中作为预处理。PD-98059(ERK1/2抑制剂)在应用EDA前加入培养基中并作用1h,以抑制ERK1/2的磷酸化。CCK-8比色法检测细胞存活率;Western blot法检测总量及磷酸化ERK1/2蛋白的表达;罗丹明123(Rh123)染色荧光显微镜照相检测线粒体膜电位(MMP)。结果 80μmol/L ISO处理H9c2心肌细胞24h,可使磷酸化的ERK1/2及MMP的水平明显降低。EDA在10～40μmol/L浓度范围内预处理1h可以剂量依赖性地减弱80μmol/L ISO处理H9c2心肌细胞48h引起的毒性反应;40μmol/L EDA预处理1h可明显抑制ISO作用24h引起的ERK1/2磷酸化水平降低及MMP受损。ERK1/2抑制剂,PD-98059,可以取消EDA诱导的细胞保护作用,使细胞毒性及MMP受损加重。结论 EDA可保护H9c2心肌细胞对抗ISO诱导的损伤作用,其机制之一可能与拮抗ISO对ERK1/2的磷酸化抑制作用有关。Objective To explore the effect of extracellular signal regulated kinase 1/2 （ERK1/2） on edaravone （EDA） -triggered protection against myocardial toxicity induced by isoprenaline （ISO） in H9c2 myocardial cells （H9c2 cells）. Methods H9c2 cells were exposed to ISO at different concentrations to establish a cardiac toxicity model induced by persistent excitation of β1 receptor. EDA was added before ISO as a pretreatment. PD-98059,an ERK1/2 inhibitor,was administered 1 h prior to EDA to inhibit the phosphorylation of ERK1/2. Cell viability was measured using cell counter kit （CCK-8）. The expressions of p-ERK1/2 and t-ERK1/2 were tested by Western blotting. Mitochondrial membrane potential （MMP） was detected by Rhodamine123 （Rh123） staining and photofluorography. Results Exposure of H9c2 cells to 80 μmol/L ISO for 24 h down-regulated ERK1/2 phosphorylation and repressed MMP. Pretreatment with 10-40 μmol/L EDA for 1 h inhibited ISO-induced myocardial toxicity and pretreatment of 40 μmol/L EDA partially rescued ERK1/2 phosphorylation and MMP level. PD-98059 abolished cardiac protection of EDA,leading to myocardial toxicity and MMP loss. Conclusion EDA can protect H9c2 cells against myocardial injury induced by ISO by suppressing ISO-triggered inhibition of ERK1/2 activation.

关 键 词：细胞外信号调节激酶1/2 依达拉奉 异丙肾上腺素 细胞毒性 线粒体膜电位 

分 类 号：R363[医药卫生—病理学]