槲皮素对血管内皮细胞的作用及其机制研究

Effect of quercetin on angiogenesis and its molecular mechanism in human umbilical vein endothelial cells

目的:观察不同浓度槲皮素对人脐静脉血管内皮细胞(HUVECs)生物学特性的影响,探讨槲皮素抑制血管生成的机制。方法:应用不同浓度槲皮素处理细胞,在不同时间点,采用CCK-8法检测槲皮素对HUVECs增殖能力的影响;采用细胞划痕实验、基质胶体外成管实验检测槲皮素对HUVECs迁移及形成管腔能力的影响;采用实时荧光定量PCR检测槲皮素对HUVECs中血管内皮细胞因子(VEGF)、碱性成纤维细胞生长因子(bFGF)mRNA表达的影响;采用Western免疫印迹法检测HUVECs中AKT信号通路相关蛋白表达的变化。采用SPSS 17.0软件包对数据进行单因素方差分析和Tukey post-hoc分析。结果:槲皮素能抑制HUVECs的增殖;与对照组相比,槲皮素处理后,HUVECs细胞迁移及形成管腔结构的数量明显减少,抑制作用呈剂量依赖性。槲皮素能抑制HUVECs中VEGF、bFGF m RNA的表达,抑制HUVECs中AKT相关信号通路。结论:槲皮素抑制HUVECs增殖及体外血管形成能力,其机制部分通过AKT相关信号通路介导。

PURPOSE：To investigate the effect of quercetin on angiogenesis and the biological characteristics of human umbilical vein endothelial cells（HUVECs）. METHODS： HUVECs were treated with quercetin（1,5 and 10 μmol/L）.CCK8 assay was used to assess the effect of quercetin on HUVECs proliferation. Wound healing assay and Matrigel-based tube formation assay were applied to observe migration and tube formation of HUVECs. Real-time PCR was used to evaluate gene expressions of VEGF and bFGF in HUVECs. Western blot was preformed to detect the change of AKT signaling pathway. Differences between groups were analyzed with SPSS 17.0 software package for ANOVA and Tukey post hoc analysis. RESULTS： Compared with the control group, quercetin significantly suppressed HUVECs proliferation,migration and tube formation in a dose-dependent manner. Gene expression of VEGF and bFGF in quercetin treated groups was significantly less than in the control group. Western blot indicated that quercetin inhibited AKT signaling pathway in HUVECs in vitro. CONCLUSION： Quercetin suppresses proliferation and angiogenesis of HUVECs in vitro,the potential mechanism of the inhibition partly involves in blockage of AKT signaling pathway.