芸香宁碱抑制血管生成作用的研究

目的：研究芸香宁碱对血管生成的抑制作用，并研究其初步的作用机制。方法用人脐静脉内皮细胞（HUVEC）的生长、迁移实验及体内动物模型－鸡胚绒毛尿囊膜法（CAM 法）研究化合物的体内外抗血管生成作用；用酶联免疫吸附法检测芸香宁碱在非凋亡剂量时对肿瘤细胞培养上清液中 VEGF 蛋白分泌量的影响。结果芸香宁碱对血管内皮细胞具有优先抑制作用，对 HUVEC 的半数抑制剂量为（33．2±0．4）μmol·L －1，而对其他肿瘤细胞的 IC50值均大于这一数值；芸香宁碱在体外能够明显抑制内皮细胞的迁移和对细胞外基质黏附作用，并呈剂量依赖性，体内实验显示30μmol·L －1剂量浓度时显著抑制 CAM新生血管形成；芸香宁碱在15μmol·L －1和30μmol·L －1的浓度剂量时显著抑制人肝癌 HepG2细胞培养上清液中VEGF 蛋白的分泌，具有显著性差异。结论芸香宁碱在非凋亡浓度剂量具有抑制新生血管形成作用，其机制与抑制血管内皮细胞增殖以及抑制肿瘤细胞表达 VEGF 有关。

The experimental study on the anti-angiogenic effects of graveolinine

Objective To investigate the inhibitory effects and the mechanism of action of graveolinine on angiogenesis.Methods The in vitro and in vivo anti-angiogenic effects of graveolinine were tested on models of angiogenesis:proliferation,migration and chick em-bryo chorioallantoic membrane (CAM)assays.VEGF protein level in supernatant of tumour cells was determined by ELISA (enzyme-linked immunosorbent assay).Results Graveolinine inhibited cell proliferation in HUVECs and some tumour cells,but the IC50 was much lower in HUVECs (33.2 ±0.4 μmol·L -1 )than that in tumor cells (≥33.2 ±0.4 μmol·L -1 ).And graveolinine showed a remarkable inhibitive effect against the migration,adhesion of HUVEC and significant anti-angiogenesis activities in CAM assay.Also graveolinine at 15 μmol·L -1 and 30 μmol·L -1 significantly inhibited VEGF secretion in supernatants of cultured HepG2 cells.Con-clusions Graveolinine inhibits angiogenesis at non-apoptosis-inducing dosage both in vitro and in vivo.Its anti-angiogenic effect might be mediated by inhibiting endothelial cells proliferation as well as VEGF expression in tumor cells.