和厚朴酚抗血管生成作用的实验研究  

The experimental study on the anti-angiogenenic effect of honokiol

作　　者：王嘉[1] 王弢[2,3] 王自强[1] 陈菲[2] 覃文新[2] 

机构地区：[1]华东师范大学生命科学学院生物医学系 [2]上海交通大学医学院,上海市肿瘤研究所 [3]复旦大学医学院,上海200032

出　　处：《肿瘤》2007年第7期527-530,共4页Tumor

摘　　要：目的:本研究通过检测和厚朴酚在体内对鸡胚尿囊膜(chicken chorilallantoic membranes,CAM)血管生成的抑制作用及体外对肿瘤血管生成的影响,探讨和厚朴酚的抗血管生成作用及其初步机制。方法:(1)用四甲基偶氮唑盐(MTT)法检测和厚朴酚对人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)、人成纤维细胞及人结肠癌RKO细胞增殖的抑制作用;(2)采用鸡胚绒毛尿囊膜模型,观测和厚朴酚对CAM新生血管生成的抑制作用;(3)逆转录聚合酶链反应(RT-RCR)方法检测和厚朴酚对RKO细胞血管内皮生长因子A(VEGF-A) mRNA表达及细胞培养上清液VEGF蛋白分泌的影响。结果:和厚朴酚对血管内皮细胞具有优先抑制作用,对HUVEC的半数抑制剂量(inhibitory concentration 50%,IC50)为14.5μmol/L,而对原代培养的人成纤维细胞的IC50高达150μmol/L,对人结肠癌RKO细胞的IC50为42μmol/L;和厚朴酚0.1μg/只和0.2μg/只剂量时显著抑制CAM新生血管形成,抑制率分别为58%和86%;和厚朴酚在10μmol/L和20μmol/L剂量时显著抑制RKO细胞的VEGF-A mRNA表达及细胞培养上清液中VEGF蛋白的分泌,具有显著性差异。结论:和厚朴酚在非凋亡剂量时具有抗血管形成作用,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。Objective： This study was designed to evaluate the in vivo anti-angiogenic effect of honokiol in chick chorilallantoic membrances （CAM）, as well as its in vitro effect in human umbilical vein endothelial cells （HUVECs） and colorectal RKO cell line. Methods： HUVECs were cultured primarily in vitro, and the influence of honokiol on the proliferation of HUVECs, human fibroblasts and human colorectal RKO cells were evaluated by MTT assay. CAM model was used to check the inhibitory effect of honokiol on neovascularization in vivo. Vascular endothelial growth factor A （VEGF-A） mRNA expression was detected by reverse transcriptase polymerase chain reaction （RT-PCR） assay and VEGF protein level in supernatant was determined by enzyme-linked immunosorbent assay （ELISA）. Results： Honokiol inhibited cell proliferation in HUVECs and RKO cells, but the inhibitory concentration 50% （ IC50 ） was much lower in HUVECs （14.5 μmol/L） than that in primary human fibroblasts （150 μmol/L） and RKO cells （42 μmol/L）. In CAM assay, honokiol dose-dependently inhibited the neovascularization, with inhibitory rate of 58% and 86% at the dosage of 0. 1 μg and 0.2 μg per cell respectively. Honokiol at 10 μmol/L and 20 μmol/L significantly inhibited VEGF-A mRNA expression and VEGF secretion in supernatants of cultured RKO cells. Conclusion： Honokiol inhibites 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.

关 键 词：和厚朴酚 血管生成抑制剂 血管内皮生长因子类 绒毛尿囊膜 

分 类 号：R972.9[医药卫生—药品]