过氧化氢诱导人脐静脉内皮细胞株的凋亡及其调控的研究

Studies on apoptosis induced by hydrogen peroxide in human umbilical vein endothelial cells

目的 探讨过氧化氢 (H2 O2 )对人脐静脉内皮细胞株 (ECV 30 4)的损伤作用及其可能机制。方法 对体外培养的ECV 30 4,采用形态学、琼脂糖凝胶电泳及流式细胞术等方法 ,观察过氧化氢的损伤方式及调控基因蛋白Fas和bcl 2的影响。结果 一定剂量 ( 10 0、30 0、5 0 0mmol/L)的过氧化氢刺激后 ,内皮细胞发生凋亡 ,凋亡率分别是 8.38± 2 .19、14 .60± 2 .75、2 0 .63± 3.89,与对照组 2 .38± 0 .96相比 ,差异具有非常显著性 ( P <0 .0 1)。同时Fas蛋白显著增加 ,分别为17.0 3± 5 .5 9、2 0 .38± 7.0 2、2 7.60± 4.5 5 ,bcl 2蛋白明显下调 ,分别为 19.35± 3.36、18.75± 3.33、13.0 5± 2 .93,与对照组相比差异均具有显著性 (P <0 .0 5或P <0 .0 1)。结论 一定剂量的过氧化氢对血管内皮细胞的损伤可通过诱导Fas表达增加 ,bcl 2表达下调 ,而致细胞凋亡。

Objective To study the damage indu ced by hydrogen peroxide on human umbilical vein endothelial cells (HUVECs) and possib le mechanism. Methods Cultured vascular endothelial cells (VECs) were treated with H_2O_2 of various concentrations. Light microscopy, agarose gel e lectrophoresis of DNA and flow cytometer were employed to detect and observe cel l injury and expression of Fas and bcl-2 protein. Results H_2O_2 was able to induce apoptosis in HUVEC s with the rate of apoptosis being 8.38±2.19, 14.60±2.75, 20.63±3.89 in H_2O _2 groups with different concentrations of 100,300,500mmol/L respectively, s ignificantly higher than in the control group (2.38±0.96,P<0.01).Fas pr otein expression was 17.03±5.59, 20.38±7.02, 27.60±4.55 respective ly in the H_2O_2 groups, significantly higher than in the control group (7.98±1.38, P<0.05 or P<0.01). bcl-2 protein expression was 19.35±3.36, 18.75±3.3 3, 13.05±2.93 respectively in the H_2O_2 groups, significantly lower than in the control group (24.90±2.95,P<0.05 or P<0.01). Conclusion H_2O_2 could result in apoptosis of vascul ar endothelial cells by up-regulation of Fas expression and down-regulation of bcl-2 protein.