甜橙黄酮对斑马鱼及人脐静脉内皮细胞抗血管新生活性的作用

目的:观察甜橙黄酮在斑马鱼及人脐静脉内皮细胞(HUVECs)上的抗血管新生能力,并探讨其相关作用机制。方法:以150 nmol/L内皮细胞生长因子受体酪氨酸激酶抑制剂(VRI)为阳性对照组、以相应浓度DMSO为空白对照组,观察不同浓度(3,10,30μmol/L)甜橙黄酮对转基因斑马鱼血管系统的影响,并通过实时荧光定量PCR检测相关血管新生基因表达的影响;分别采用XTT与流式细胞术检测3～100μmol/L甜橙黄酮对20 ng/ml内皮细胞生长因子(VEGF)诱导的HUVECs增殖和细胞周期的影响。结果:甜橙黄酮可以剂量依赖性地抑制斑马鱼节间血管(ISV)的形成,下调血管新生相关基因hras、kdrl和vegfaa的表达;可剂量依赖性抑制VEGF诱导的HUVECs增殖,并诱导HUVECs停滞于细胞周期G0/G1期。结论:甜橙黄酮可能通过阻滞细胞周期和影响相关基因的表达达到在斑马鱼体内模型和HUVECs体外模型的抗血管新生作用。

Effects of Anti-angiogenesis Activity of Sinensetin on Human Umbilical Vein Endothelial Cells and Zebrafish

Objective: To investigate the anti-angiogenesis activity of sinensetin on human umbilical vein endothelial cells(HUVECs) and zebrafish,and to explore the underlying mechanism of the anti-angiogenesis activity.Methods: The positive control group was treated by 150 nmol/L VEGF receptor kinase inhibitor and blank control group was treated by 150 nmol/L DMSO.The effect of sinensetin at different concentrations on zebrafish’s vascular system was observed,and the expression of angiogenesis gene was analyzed by real-time PRC;proliferation of HUVECs induced by VEGF and cell cycle were detected by XTT abd FCM.Results: Sinensetin dose-dependently caused regression in intersegmental vessel(ISV) and down-regulated mRNA expressions of hras,kdrl and vegfaa in zebrafish;It also inhibited HUVEC proliferation by inducing cell-cycle arrest at the G0/G1 phase.Conclusion: New anti-angiogenesis activity of sinensetin may result in cell-cycle arrest and expression of numerous angiogenesis genes in HUVECs and zebrafish.