血管内皮生长因子对过氧化氢诱导的内皮细胞凋亡的影响及其机制

Mechanisams of Vascular Endothelial Growth Factor on Apoptosis Induced by Hydrogen Peroxide in Vascular Endothelial Cell

目的研究血管内皮生长因子对过氧化氢诱导的血管内皮细胞凋亡的影响,同时检测凋亡相关基因Bcl-2 mRNA与Fas mRNA表达的变化。方法体外培养人脐静脉内皮细胞,随机分成对照组、过氧化氢组和过氧化氢+血管内皮生长因子组,12 h后,采用原位末端标记法和流式细胞仪分别观察各组细胞的凋亡数和凋亡率,通过逆转录聚合酶链反应观察各组细胞中凋亡相关基因Bcl-2 mRNA与Fas mRNA表达的变化。结果过氧化氢组凋亡细胞数(27.83±2.14)明显高于对照组(2.50±1.05)(P<0.01)和过氧化氢+血管内皮生长因子组(13.00±2.10)(P<0.01)。过氧化氢组细胞凋亡率(14.17%±0.45%)明显高于对照组(1.55%±0.87%)(P<0.01)和过氧化氢+血管内皮生长因子组(5.69%±0.38%)(P<0.01)。与过氧化氢组比较,过氧化氢+血管内皮生长因子组Fas mRNA表达明显减少(40.67%±2.16%比94.50%±3.45%,P<0.01),而Bcl-2 mRNA表达明显增加(60.33%±1.75%比23.17%±1.17%,P<0.01)。结论血管内皮生长因子能拮抗过氧化氢诱导的内皮细胞凋亡,其抗凋亡的机制可能与上调Bcl-2 mRNA表达与下调Fas mRNA表达有关。

To observe the effect of vascular endothelial growth factor （VEGF） on the apoptosis and the expression of Fas mRNA and Bcl-2 mRNA in human umbilical vein endothelial cell （hUVEC ）. Methods hUVEC were randomly divided into three groups： control group, hydrogen peroxide group and hydrogen peroxide ± VEGF group. After 12 hours, the apoptosis of hUVEC was determined by flow cytometry （FCM） and terminal deoxynueleofidyl transferase biofin-dUTP nick end labeling （TUNEL）, the expression of Be1-2 mRNA and Fas mRNA were examined by reverse transcription polymerase chain reaction（RT- PCR）. Results Apoptosis number in hydrogen peroxide group was higher than that in control group （27.83±2.14 vs 2.50 ±1.05, P〈0.01）. However, inhydrogen peroxide±VEGF group, apoptosis number was lower than that in hydrogen peroxidegroup（13.00±2.10 vs 27.83±2.14, P〈0.01）. Apoptosis rate in hydrogen peroxide group was higher than that in centrol group （ 14.17% ± 0.45% vs 1.55% ± 0.87%, P 〈 0.01 ） and in hydrogen peroxide ± VEGF group（ 14.17% ± 0.45% vs 5.69% ± 0.38%, P 〈 0.01 ）. Compared with control group, hydrogen peroxide markedly increased Fas mRNA expression （94.50% ± 3.45% vs 21.17% ± 1.17%, P 〈 0.01） and decreased Bel-2 mRNA expression （23.17% ± 1.17% vs 85.17% ±1.47%, P 〈 0.01）. Compared with hydrogen peroxide group, VEGF significantly decreased Fas mRNA expression （40.67% ± 2.16% vs 94.50% ± 3.45%, P 〈 0.01 ） and increased Bcl-2 mRNA expression（60.33% ± 1.75% vs 23.17% ± 1.17%, P 〈0.01）. Conclusions VEGF can inhibit the apoptosis induced by hydrogen peroxide in HUVEC, which may correlated with upregulation Bcl-2 mRNA expression and inhibiting Fas mRNA expression.