过氧化氢诱导大鼠主动脉内皮细胞氧化损伤模型的建立  

Establishment of Oxidative Injury Model of SVAREC in Vitro

作　　者：黄晶晶[1] 施晓峰[1] 陈宇驰[1] 夏威标 金波[1] 丁志山[1] 

机构地区：[1]浙江中医药大学生命科学学院,浙江杭州310053

出　　处：《云南中医学院学报》2015年第3期5-9,共5页Journal of Yunnan University of Traditional Chinese Medicine

基　　金：中国博士后科学基金(2014M550335)

摘　　要：目的采用过氧化氢(H2O2)诱导大鼠主动脉内皮细胞(SVAREC)氧化损伤,建立SVAREC氧化损伤模型。方法体外培养SVAREC细胞,绘制细胞生长曲线,用不同浓度的H2O2刺激细胞,并在不同时间点用MTS法测定细胞存活率,以确定SVAREC氧化损伤模型条件。倒置显微镜下观察细胞形态学变化,微孔酶标法测乳酸脱氢酶(LDH)活力,流式细胞术检测细胞凋亡率,WB检测细胞中半胱氨酸蛋白酶(Caspase-3)的表达情况。结果与空白对照组细胞比较,当H2O2浓度为100μM,作用时间为2 h时,造成细胞损伤,其细胞凋亡率和Caspase-3蛋白表达量明显高于空白对照组。结论该实验条件下H2O2浓度为100μmol/L,作用时间为2 h是模型的最佳复制条件。Objective To establish the oxidative damage model of SVAREC. Methods SVAREC were cultured in vitro and the growth curves were draw. Different concentration of H2O2 were stimulated the cells,and the cell viability were measured by the method of MTS at different time. Under the condition of this model,cellular morphology were observed by inverted microscope,the activity of LDH were measured by microplate reader,the apoptosis rate of cells also were detected by flow cytometry,and Caspase-3 expression were tested by Western blotting. Results Compared with control,cells were injured at the concentration of H2O2 is 100μmol/L for 2 h,which,the apoptosis rate and protein expression of Caspase-3 were significantly higher. Conclusion The best replication model of this experimental condition is exposure the SVAREC at the 100μmol/L concentration of H2O2 for 2 h.

关 键 词：内皮细胞 氧化应激 过氧化氢 细胞损伤 凋亡 

分 类 号：R285.5[医药卫生—中药学]