衰老对AhR 及其血管生成相关因子表达的影响

目的:探讨芳香烃受体(aryl hydrocarbon receptor,Ah R)在过氧化氢(H_2O_2)诱导的人脐静脉内皮细胞(HUVECs)衰老模型中表达的变化情况及其对血管生成相关因子表达的影响。方法:采用不同浓度的H_2O_2(100μM,200μM,300μM)处理HUVECs诱导内皮细胞衰老,通过β-半乳糖苷酶染色观察细胞衰老情况,Western blot检测HUVECs中Ah R蛋白以及血管生成相关因子低氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)蛋白表达,ELISA检测细胞培养上清液中血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的变化。结果:与对照组相比,经过不同浓度H_2O_2处理后,HUVECs均出现细胞衰老表现,Ah R蛋白表达显著增加,HIF-1α以及VEGF蛋白表达明显减少,且均呈现出剂量依赖性,以300μM H_2O_2处理组最为显著。结论:在过氧化氢诱导衰老的人脐静脉内皮细胞中Ah R蛋白表达明显增加,HIF-1α和VEGF等促血管生成因子明显减少。

Effects of Aging on the Expression of AhR and Angiogenesis Related Factors

Objective:To investigate the expression change of aryl hydrocarbon receptor (aryl hydrocarbon receptor, AhR) in hydrogen peroxide (H2O2) induced senescence and AhR effects on angiogenesis related factor in senescent human umbilical vein endothelial cells (HUVECs).Methods:Senescence induced by different concentrations of H2O2 (100 uM, 200 uM, 300 uM) in HUVECs was examined by beta-galactosidase staining. Protein expression of AhR and hypoxia-inducible factor-1-alpha(HIF-1alpha) were analysed byWestern blot. The concentrations of vascular endothelial growth factor (VEGF) in culture medium were tested by ELISA.Results:Compared with the control group, all of the H2O2 groups showed signs of senescence. Meanwhile, H2O2 significantly increased the AhR protein expression and decreased the protein expression of HIF-1alpha and VEGF in a dose-dependent manner in HUVECs. Those changes were most obviously in 300uM treatment group.Conclusion:Protein expression of AhR up regulated in H2O2-induced senescent HUVECs, as well as angiogenesis related factors (HIF-1alpha, VEGF) decreased significantly.