H_2O_2、LPS和TNF-α诱导人类脐静脉内皮细胞损伤的实验研究

H_2O_2、LPS and TNF-αinduced injury of human umbilical vein endothelial cells

目的探讨人类脐静脉内皮细胞损伤模型建立方法,为体外研究血管内皮细胞提供实验基础。方法分离培养人类脐静脉内皮细胞,分别采用不同浓度的过氧化氢(H2O2)、脂多糖(LPS)、肿瘤坏死因子(TNF)-α刺激细胞,孵育不同时间后,采用四甲基偶氮唑盐(MTT)法检测细胞活力(OD值)。结果各浓度H2O2损伤组较对照组OD值显著降低(P<0.01),不同浓度H2O2损伤组间OD值无统计学差异(P>0.05)。0.1μg/mL LPS损伤组与对照组OD值无统计学差异(P>0.05);其余各浓度LPS损伤组较对照组OD值显著降低(P<0.05),且LPS浓度在一定范围内,OD值随LPS浓度的增加及作用时间的延长而降低,具有一定的浓度与时间依赖性。各浓度TNF-α损伤组较对照组OD值显著降低(P<0.01),且TNF-α浓度在一定范围内,OD值随TNF-α浓度的增加及作用时间的延长而降低,具有一定的浓度与时间依赖性。结论 H2O2、LPS和TNF-α能体外损伤人类脐静脉内皮细胞,成功建立人类脐静脉内皮细胞损伤模型,且在一定浓度范围内各损伤因子对人类脐静脉内皮细胞损伤程度呈浓度和时间依赖性。

Objective To investigate the method of establishing human umbilical vein endothelial cells （HUVECs） injury model ,and to provide experimental basis for the research of vascular endothelial cells in vitro . Methods HUVECs were separated and cultured ,the hydrogen peroxide （H2O2 ） ,lip polysaccharide （LPS） and tumor necrosis factor‐alpha （TNF‐α） with different concentrations were used to induce the cell damage .MTT assay was used to determine the vitality of cells （OD value）at different time interval .Results OD value of H2O2 damage groups was significantly lower than that of normal control group （P〈 0 .01） .No significant differences were found among H2O2 damage groups （P〉 0 .05） .No significant differences were found between 0 .1 μg/mL LPS damage group and normal control group （P〉0 .05） .OD value of the other LPS damage groups was significantly lower than that of normal control group （P〈 0 .05） .The effect of LPS on HUVECs was time and concentration dependent within a certain range .OD value of TNF‐α damage groups was significantly lower than that of normal control group （P〈 0.01）.The effect of TNF‐α on HUVECs was time and concentration dependent within a certain range. Conclusion The injury model of HUVECs could be induced by H2O2 ,LPS and TNF‐αin vitro .