白头翁联合恩度体外抗血管生成作用

The effect of Pulsatillae combined with Endostar on the proliferation,migration,build of tube of vessel endothelial cells

目的:探讨白头翁醇提物联合恩度体外对血管内皮细胞增殖、迁移、小管形成的作用。方法:以人结肠癌Lovo细胞为对照,采用MTT法观察中药白头翁联合恩度对人脐静脉内皮细胞(HUVEC)体外增殖的影响;利用transwell小室趋化实验、体外小管形成实验观察中药白头翁联合恩度对HUVEC体外迁移、成血管能力的影响。结果:白头翁醇提物2-8μg/ml在体外具有抑制HUVEC增殖的作用,联合恩度10μg/ml时具有协同作用(细胞存活率在37.6%-22.5%),与浓度呈负相关(r=-0.850,P=0.032),此浓度范围内对人结肠癌Lovo细胞增殖的抑制明显低于对HUVEC的抑制,具有明显差异细胞毒作用(P=0.000)。白头翁醇提物2-8μg/ml联合恩度10μg/ml作用12h后具有抑制HUVEC迁移的作用,与浓度呈负相关(r=-0.919,P=0.000)。白头翁醇提物2-8μg/ml联合恩度10μg/ml作用24h后内皮细胞小管数目减少,不能相互连接成网状结构,而且管腔不完整,与对照组相比有显著性差异,与浓度呈负相关(r=-0.908,P=0.000)。结论:小剂量白头翁在体外具有抑制血管生成作用,联合恩度具有协同效应。

Objective：To study the effect of pulstatissal combined with Endostar on the proliferation,migration,build of tube of vessel endothelial cells.Methods：Proliferations of human umbilical vein endothelial cells（HUVEC） were measured by MTT colorimetric assay treated by Pulsatillae combined with Endostar in vitro compared with human colon carcinoma Lovo cell.Using transwell cabin test,canaliculization test to observe the effect of Pulsatillae combined with Endostar on the migration,vaso-formed ability.Results： The survival rate of HUVEC was 37.6%-22.5% within the dose of 2-8μg/ml and negative correlated with the concentration（r=-0.850,P=0.032） treated with the alcohol extract of Pulsatillae.The survival rate of Lovo cell was more than HUVEC,so the alcohol extract of Pulsatillae had differential cytotoxicity to Lovo cell and HUVEC（P=0.000）.And there was a synergistic effect when combined with Endostar 10μg/ml.The migration of HUVEC was obviously depressed treated with the alcohol extract of Pulsatillae at the doe of 2-8μg/ml combined with Endostar 10μg/ml in 12 hours,and negatively correlated with the concentration（r=-0.919,P=0.000）.The number of tubule was reduced and lumina become unintegrated,there was significant difference compared with the control group treated with Pulsatillae 2-8μg/ml combined with Endostar 10μg/ml,and negatively correlated with the concentration（r=-0.908,P=0.006）.Conclusion： Low dose Pulsatillae can actively suppress angiogenesis in vitro,and has synergistic effect with Endostar.