过氧化氢诱导HUVECs氧化应激模型的构建

戴青里; 孙贵虎; 闫斌; 郭涛; 戴青原

过氧化氢诱导HUVECs氧化应激模型的构建

1. 大理白族自治州人民医院超声科
2. 昆明医科大学第一附属医院心内科
3. 昆明医科大学第一附属医院门诊部

基金项目:

基金：云南省教育厅科学研究基金资助项目 (2015Y153);

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出版历程
- 收稿日期: 2018-02-04

Establishment of Oxidative Stress Model by Inducing Human Umbilical Vein Endothelial Cells Using Hydrogen Peroxide

Funds:

基金：云南省教育厅科学研究基金资助项目 (2015Y153);

摘要

摘要: 目的利用过氧化氢 (H2O2) 诱导人脐静脉内皮细胞 (human umbilical vein endothelial cells, HUVECs) 构建体外氧化应激细胞模型.方法 H2O2分6个浓度梯度处理HUVECs不同时间, 倒置显微镜观察细胞形态变化, CCK-8法检测细胞存活率, 流式细胞术检测细胞凋亡率, DCFH-DA荧光探针和流式细胞仪检测细胞内活性氧水平, 筛选H2O2的最佳作用浓度和时间.结果不同浓度H2O2处理细胞不同时间, 对HUVECs均有损伤作用;400、600、800μmol/L H2O2处理HUVECs 24 h, 细胞存活率显著降低 (P<0.05) , 800μmol/L H2O2作用HUVECs不同时间, 细胞存活率随处理时间延长逐渐降低 (P<0.01) ;各组H2O2处理HUVECs 24 h, 细胞凋亡率显著增加 (1 000μmol/L除外, P<0.05) , 坏死率亦显著增加 (100μmol/L除外, P<0.05) ;800μmol/L H2O2处理HUVECs不同时间, 细胞凋亡率随处理时间延长逐渐增加, 24 h时达到最高 (P<0.05) , 细胞坏死率亦逐渐增加, 48 h时达到最高 (P<0.05) ;各组H2O2作用HUVECs 24 h, 浓度400μmol/L时, 细胞内ROS水平达到最高 (P<0.01) , 800μmol/L H2O2处理HUVECs, 胞内ROS水平随处理时间延长而递增 (P<0.01) .结论800μmol/L H2O2与HUVECs作用24 h可构建体外细胞氧化应激模型.
- 人脐静脉内皮细胞 /
- 过氧化氢 /
- 氧化应激 /
- 细胞模型
Abstract: Objective To establish an oxidative stress cell model by inducing human umbilical vein endothelial cells (HUVECs) using hydrogen peroxide (H2 O2) . Methods Six gradient concentrations of H2 O2 were co-cultured with HUVECs for 6 h, 12 h, 24 h and 48 h. Inverted microscope was used to observe the change of cell morphologies. The optimal working concentration and effect time of H2 O2 on HUVECs were screened by CCK-8 method for cell viability, flowcymetry for apoptosis rate, and CDFU-DA fluorescent probing and flowcymetry for ROS levels. Re s ults Any concentration of H2 O2 could do harm to HUVECs co-culturing with cells. Cell viabilities decreased statistically when treating cells with 400, 600 or 800 μmol/L H2 O2 for different times (P<0.05) . Cell viabilities also decreased gradually as time prolonged when treating cells with 800 μmol/L H2 O2 (P<0.001) . When treating cells with different concentration of H2 O2 for 24 h, cell apoptosis rates increased dramatically (1000μmol/L as an exception, P<0.05) , so did cell necrosis rates (100 μmol/L as an exception, P<0.05) ;When treating cells with 800 μmol/L H2 O2, cell apoptosis rates increased dramatically as time prolonged, reaching the peak at 24 h (P<0.05) , so did necrosis rates, reaching the peak at 48 h (P<0.05) . ROS level reached thepeak while treating cells with 400 μmol/L H2 O2 (P<0.001) ; ROS levels increased gradully as time prolonged when treating cells with 800 μmol/L H2 O2 (P<0.001) . Conclusions The oxidative stress model in HUVECs was established by coculturing cells with culture medium containing 800 μmol/L hydrogen peroxide for 24 h.
- Human umbilical vein endothelial cells /
- Hydrogen peroxide /
- Oxidative stress /
- Cell model

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参考文献(16)

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