摘要
目的观察H2O2作用下乳鼠心肌细胞早期凋亡和细胞内氧化还原状态的改变及淫羊藿总黄酮注射液(epimedium flavonoids injection,EFI)的干预作用。方法体外培养大鼠乳鼠心肌细胞,复制H2O2损伤模型,流式细胞仪检测心肌细胞早期凋亡和活性氧水平,测定心肌细胞内总抗氧化能力,免疫印迹法检测心肌细胞硫氧还蛋白(thioredoxin,Trx)和超氧化物歧化酶(superoxide dismutase,SOD)蛋白表达水平。结果与正常对照组比较,100μmol·L-1H2O2作用12h后心肌细胞早期凋亡率显著增高(P<0.001);心肌细胞平均荧光强度(MFI)明显上升(P<0.01);总抗氧化能力显著下降(P<0.01或P<0.05);Trx和SOD蛋白表达减少。EFI可剂量依赖性降低心肌细胞早期凋亡率和MFI、升高总抗氧化能力、Trx和SOD蛋白表达增加,与H2O2作用组比较,200和400mg·L-1EFI组差异显著(P<0.05或P<0.01)。结论H2O2可诱导心肌细胞早期凋亡,促使心肌细胞处于氧化状态;EFI可促使心肌细胞处于还原状态,对H2O2诱导的心肌细胞损伤具有保护作用。
OBJECTIVE To explore the effects of EFI on early apoptosis and redox status in neonatal rat cardiomyocytes. METHODS Ventricular myocytes were isolated from neonatal rat and cultured. The oxidative stress model for neonatal rat cardiomyocytes was made by H2O2, Earlyapoptosis rates and ROS levels were detected by flow cytometer. Total anti-oxidative abilities were measured. The protein expressions of thioredoxin（Trx） and superoxide dismutase（SOD） were observed by western blotting. RESULTS After administrating 100 μmol·L^-1 H2O2 for 12 h, the early apoptosis rates were enhanced significantly （P 〈 0. 001 ）, mean fluorescence intensity（MFI） was increased obviously （ P 〈 0. 01 ）, total anti-oxidative ability was decreased remarkably （ P 〈 0. 01 or P 〈 0.05） , Trx and SOD protein levels were reduced. EFI dose-dependently decreased early apoptosis rates and MFI, enhanced total anti-oxidative abilities,Trx and SOD protein levels were increased as well. Compared with H2O2 group, there were different significantly between the groups treated with 200 and 400 mg ·L^-1 EFI （P 〈 0. 05 or P 〈 0. 01 ）. CONCLUSION H2O2 can induce neonatal rat cardiomyocytes to be in early apoptosis and in oxidative stress state. EFI can reduce the redox state in neonatal rat cardiomyocytes. It suggests that EFI can protect the oxidative injury in neonatal rat cardiaomyocytes induced by H2O2.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2007年第11期832-835,共4页
Chinese Pharmaceutical Journal
基金
国家民族事务委员会资助项目(05XY07)
