硒对砷诱导的大鼠H9c2心肌细胞氧化损伤的抑制作用和可能机制  

Protective effects and possible mechanisms of selenium on oxidative insults of H9c2 cardiac cells induced by arsenic

作　　者：薛晚利[1] 王治伦[1] 李选社[1] 董璐[1] 杨浩杰[1] 

机构地区：[1]西安交通大学地方病研究所,西安710061

出　　处：《中国地方病防治》2010年第2期84-88,共5页Chinese Journal of Control of Endemic Diseases

基　　金：国家自然科学基金资助项目<砷对大鼠卵巢毒作用分子机制和相关蛋白质组学的研究>(30872188)

摘　　要：目的探讨硒对砷诱导的H9c2心肌细胞氧化损伤的保护作用及其可能机制。方法H9c2细胞按处理方法不同分对照组、Na2SeO3(1μmol/L)组、As2O3(5μmol/L)组、As2O3(5μmol/L)+Na2SeO3(1μmol/L)组以及NF-κB抑制剂PDTC(100 nmol/L)不同处理组。应用MTT法检测细胞存活率;DTNB法检测细胞内还原型谷胱甘肽(GSH)含量;二氢乙啶(Dihydroethidium,DHE)染色法检测细胞活性氧(reactive oxygen species,ROS)产生;提取细胞核蛋白,Western blotting法检测核蛋白中NF-κB的p65亚单位表达。结果与对照组相比,5μmol/L As2O3作用下,H9c2细胞生存率、细胞内GSH含量降低,ROS生成和核蛋白中NF-κB p65的表达增加;1μmol/L Na2SeO3可以提高As2O3损伤的H9c2细胞存活率和细胞内GSH含量,减少ROS生成和p65的表达。PDTC干预可减轻砷所致心肌细胞的损伤,表现为细胞ROS产生减少;Na2SeO3和抑制剂PDTC共同作用时,H9c2细胞生成ROS进一步减少。结论NF-κB信号途径参与了As2O3诱导的H9c2心肌细胞氧化应激反应,Na2SeO3可能通过抑制NF-κB减轻氧化应激诱导的H9c2细胞损伤。Objective To investigate the protective effects and possible mechanisms of selenium on oxidative insults of H9c2 cardiac cells induced by arsenic.Methods H9c2 cells were classified as four experimental groups giving different treatment： Control group,Na2SeO3（1 μmol/L）group,As2O3（5 μmol/L） group,As2O3（5 μmol/L） ＋ Na2SeO3（1 μmol/L）group and NF-κB inhibitor PDTC（100 nmol/L） combined with different groups.The cell viability was measured by MTT assay;GSH content was detected by DTNB method;Dihydroethidium（DHE） staining was used to evaluate in situ concentration of reactive oxygen species（ROS）;The expressions of NF-κB p65 were determined by western blotting.Results Compared with control group,cell viability and GSH content were decreased;ROS production and expression of p65 in nuclear were increased in As2O3 group group.Compared with As2O3 group,cell viability and GSH content were increased;ROS production and expression of p65 were decreased in As2O3 ＋ Na2SeO3 group.Meanwhile,studies showed that combined effects of PDTC significantly decreased the oxidative insults in As-induced H9c2 as judged by the ROS production measurements.When Na2SeO3 treatment was performed in the presence of PDTC,there was a further reduction in ROS production.Conclusion NF-κB pathway involved in the arsenic-induced oxidative stress of H9c2 cardiac cell.Na2SeO3 probably attenuated the oxidative insults by suppressing NF-κB pathway.

关 键 词：硒 砷 氧化应激 NF-ΚB H9C2细胞 

分 类 号：O613.52[理学—无机化学] O613.63[理学—化学]