枸橘苷抑制血管生成作用的研究

The experimental study on the anti-angiogenic effects of poncirin

目的研究传统中药枸橘主要成分之一枸橘苷对血管生成的抑制及初步的作用机制。方法用四甲基偶氮唑盐(MTT)法、黏附抑制试验、迁移抑制实验及体内鸡胚绒毛尿囊膜法(CAM法)研究枸橘苷体外抗血管生成作用;用逆转录聚合酶链反应(RT-RCR)方法检测枸橘苷对肿瘤细胞血管内皮生长因子A(VEGF-A)mRNA表达的影响。结果枸橘苷对血管内皮细胞增殖表现出良好的抑制作用,IC50值为(27.8±0.2)μmol·L-1,优于其抑制肿瘤细胞增殖的能力[IC50≥(33.8±0.1)μmol·L-1];在体外该化合物能够明显抑制内皮细胞的黏附和迁移作用,同时体内实验显示该化合物在30μg剂量浓度时能够显著抑制鸡胚绒毛尿囊膜新生血管形成;RT-RCR实验显示枸橘苷在12.5μmol·L-1和25μmol·L-1剂量时显著抑制A549细胞的VEGF-A mRNA表达,且结果具有显著的差异。结论枸橘苷具有抑制新生血管形成作用,其作用机制可能与与抑制血管内皮细胞增殖以及降低肿瘤细胞表达VEGF的量有关。

Objective This s tudy was to investigate the inhibitory effects and the mechanism of action of poncirin on angiogenesis. Methods MTT assay was used to research the effect of poncirin the proliferation of HUVECs（ human umbilical vein endothelial cells） and some human cancer cells. Adhesion inhibition experiment,migration inhibition experiment and chick embryo chorioallantoic membrane（ CAM） assays were testas models of angiogenesis. VEGF- A（ vascular endothelial growth factor A） mRNA expression was detected by RT- PCR（ reverse transcriptase polymerase chain reaction） assay. Results Poncirin inhibited cell proliferation in some tumour cells,and the IC50 was much lower in HUVECs（ 27. 8 ± 0. 2μmol·L- 1） than that in tumor cells（ IC50≥33. 8 ± 0. 1μmol·L- 1）. And poncirin showed a remarkable inhibitive effect against the migration,adhesion of HUVEC and significant anti- angiogenesis activities in CAM assay. Also poncirin at 12. 5μmol·L- 1and 25μmol·L- 1inhibited significantly inhibited VEGF- A mRNA expression in cultured A549 cells. Conclusion Poncirin could inhibit angiogensis through inhibiting endothelial cells proliferation and VEGF expression in tumor cells.