中文摘要：

探讨沙蟾毒精（arenobufagin）对体内外血管生成的抑制作用。采用MTT法检测沙蟾毒精对人鼻咽癌细胞（CNE-2）、人喉癌细胞（Hep2）、人神经母细胞瘤（SH-SY5Y）、人结肠癌细胞（LOVO）、人前列腺癌细胞（PC-3及DU145）和人脐静脉内皮细胞（HUVECs）的细胞毒性及运用光学显微镜观察细胞形态的变化，发现沙蟾毒精剂量依赖性抑制CNE-2、Hep2、SH-SY5Y、LOVO、PC-3、DU145和HUVECs增殖，且在对人癌细胞亚细胞毒浓度下能够有效抑制HUVECs的增殖。采用鸡胚尿囊膜（chick embryo chorioallantoic membrane，CAM）模型观察新生血管的生成，结果发现沙蟾毒精作用24h后，尿囊膜新生血管生成明显减少。采用细胞周期分析法观察到沙蟾毒精阻滞HUVECs于G2/M期，且随着浓度增加，细胞出现sub-G，凋亡峰。运用流式细胞术检测沙蟾毒精作用HUVECs后的凋亡现象以及线粒体膜电位的变化，确证沙蟾毒精呈时问和剂量依赖性诱导HUVECs凋亡，同时检测到线粒体膜电位下降。Westem blotting检测发现，沙蟾毒精作用HUVECs后，凋亡标志分子PARP被切割。上述研究表明，沙蟾毒精具有明显的体内外抑制血管生成作用，其抑制作用可能与诱导血管内皮细胞周期阻滞及凋亡有关。

英文摘要：

This study is to investigate the anti-angiogenetic effect of arenobufagin in vitro and in vivo. The anti-proliferation effect of arenobufagin on CNE-2, Hep2, SH-SY5Y, LOVO, PC-3 and DU 145 cells as well as human umbilical vein endothelial cells （HUVECs） was determined by MTT assay. Cell morphological changes of LOVO and HUVECs after arenobufagin treatment were observed by microscopy. Arenobufagin inhibited the proliferation of CNE-2, Hep2, SH-SY5Y, LOVO, PC-3, DU145 and HUVECs in a dose-dependent manner. Furthermore, it was obviously observed that the subcytotoxic concentration of arenobufagin in human carcinoma cells induced a marked decrease in the viability of HUVECs. Chick embryo chorioallantoic membrane （CAM） model was used to detect the anti-angiogenetic effect of arenobufagin in vivo. Arenobufagin significantly suppressed the angiogenesis of CAM. Cell cycle analysis demonstrated that G2/M phase was arrested and the sub-G1 peak appeared with the increase of arenobufagin concentration. PI/Annexin V double stainingassay further demonstrated that arenobufhgin could induce apoptosls in a dose- and time-dependent manner. Mitochondrial potential collapse detected by flow cytometric analysis was increased after arenobufagin treatment. It also observed that PARP was cleaved to p85 active form by Western blotting. Taken together, arenobufagin has significant anti-angiogenetic effect in vitro and in vivo, and the action mechanisms behind its anti-angiogenesis may be associated with cell cycle arrest and apoptosis of vein endothelial cells.