过氧化氢致H9c2大鼠心肌细胞DNA损伤作用  

Study on hydrogen peroxide induced DNA damage in myocardial H9c2 cells

作　　者：冯星[1] 孙磊[1] 郭彩霞[1] 金明华[1] 刘晓梅[1] 刘颖[1] 孙志伟[1] 

机构地区：[1]吉林大学公共卫生学院,长春130021

出　　处：《中国公共卫生》2009年第3期296-297,共2页Chinese Journal of Public Health

基　　金：教育部博士点基金(2003183006)

摘　　要：目的研究不同浓度过氧化氢(H2O2)对H9 c2大鼠心肌细胞DNA损伤及诱导细胞凋亡的作用。方法H9 c2心肌细胞处理组分别暴露于50,100,200,400μmol/L H2O2中1,3,6,12,24 h后,采用四甲基偶氮噻唑蓝(MTT)法检测细胞存活率;采用丫啶橙/溴乙锭(AO/EB)双染技术观察细胞凋亡;利用单细胞凝胶电泳技术(SCGE)观察细胞DNA损伤。结果H2O2可以引起H9 c2心肌细胞损伤,随时间呈剂量效应关系(P<0.05),其中24 h,400μmol/L剂量组细胞存活率最低达40.6%。H2O2可诱导H9 c2心肌细胞出现凋亡并观察到明显的凋亡形态学的改变,与阴性对照组比较,差异有统计学意义(P<0.05),其中6 h时,100μmol/L剂量组细胞凋亡最明显,凋亡率达22.2%。H2O2可引起H9 c2心肌细胞DNA损伤,且损伤随剂量增加而增大(P<0.01),以400μmol/L剂量组最为明显,细胞DNA损伤率可达64.0%。结论H2O2造成H9 c2心肌细胞氧化损伤和DNA损伤,随着其暴露剂量和时间的不同表现为凋亡和坏死。Objective To study the effects of hydrogen peroxide on DNA damage and apoptosis in myocardial H9c2 cells line. Methods H9c2 cells were exPosed to different concentration of H202 （50,100,200,400 μmol/L） in different time （1,3,6,12,24h）. The cell viability was measured by MTT assay. APoptosis was determined by AO/EB double fluorescent staining assay. The single cell gel electrophoresis（SCGE） was used to study the DNA damage of H9c2 cells. Results The experimental results demonstrated that H202 could damage H9c2 cells in dose dependant and time dependant way（ P 〈 0.05 ）, and induce apoptosis with obviously morphological changes in exPosed groups （ P 〈 0. 05 ）, especially at 6h, 100 μmol/L level of H2 02, compared with NC （ P 〈 0. 01 ）. H2 O2 could cause DNA damage. The rate of DNA damage was increased with the increase of dosage（P 〈 0. 01 ）. Conclusion H202 could cause oxidative injury and DNA damage in H9c2 cells. APoptosis and necrosis are main outcomes depending on exPosure dose and exPosure time.

关 键 词：过氧化氢(H2O2) H9C2心肌细胞 细胞凋亡 坏死 DNA损伤 

分 类 号：R994.4[医药卫生—毒理学]