重楼醇提物体外抑制血管生成作用研究

Inhibitory effect of paridis on angiogenesis in vitro

目的:探讨重楼醇提物对体外血管生成的抑制作用及其可能的机制。方法:采用MTT法观察重楼醇提物对人脐静脉血管内皮细胞(HUVEC)、人肠癌LOVO细胞增殖的影响。利用transwell小室趋化实验、体外管腔形成实验观察重楼醇提物对HUVEC迁移、成血管能力的影响。结果:重楼醇提物在(15-60)μg/ml浓度范围内,内皮细胞抑制率在24.2%-82.1%之间,具有抑制血管内皮细胞增殖作用;体外管腔形成实验发现,(15-60)μg/ml的重楼醇提物管腔形成数目减少,且管腔不完整,与对照组相比有显著性差异(P分别为0.0000011、0.0000417、0.0008752)。经(15-60)μg/ml的重楼醇提物处理后内皮细胞迁移数明显少于对照组(P分别为0.0000005、0.0000033、0.009993)。而在该浓度范围内,对人肠癌LOVO细胞无明显细胞毒作用(细胞抑制率在12.1%-13.9%之间)。(15-60)μg/ml的重楼醇提物可诱导内皮细胞凋亡,抑制内皮细胞DNA的合成,且呈剂量依赖性(相关系数分别为0.929、-0.922,P为0.036、0.039)。结论:重楼醇提物在体外能有效抑制血管生成。其机制可能与抑制内皮细胞增生、迁移和管腔形成,诱导内皮细胞凋亡,抑制内皮细胞DNA的合成有关。

Objective：To approach the inhibitory action and the mechanism of the alcohol extract of pafidis to angiogenesis in vitro. Methods ： The MTT assay was used to observed the effect of the alcohol extract of pafidis to EC of human umbilical vein （HUVEC） and human intestine carcinoma LOVO cell. Transwell cabin test and out - body canaliculization test were used to observe the alcohol extract of paridis impact the migrational and vaso - formed ability of HUVEC. Results： When the concentration of the alcohol extract of pafidis between 15μg/ml to 60μg/ml, the in- hibition rate of EC was between 24.2% and 82.1% , and there was inhibitory action to vascular endothelial cell proliferation. When the concentration of the alcohol extract of paridis between 15μg/ml to 60μg/ml, numbers of canaliculization was reduced and lumens was unintegrlty in out -body eanalieulization test. And there was significant difference compared with the control group （P = 0.0000011, 0. 0000417, 0. 0008752）. And the numbers of endothelial cell migration was significant reduced than the control group （P = 0.0000005, 0.0000033, 0. 009993 ）. There was no evident cell toxic action to human intestine carcinoma LOVE cell （ the cell inhibition was between 12. 1% to 13.9% ） between the range of concentration. The alcohol extract of paridis can induce endothelial cell apoptosis, suppress endothelial cell DNA synthesis, and there was dose dependent （ coefficient correlation 0.929 and - 0.922, and P = 0.036, 0.039）. Conclusion： The alcohol extract of paridis can active suppress the angiogenesis in vitro. The mechanism maybe concerned with inhibited proliferation, migration and canaliculization of endothelial cell, induced the apoptosis of endothelial cell and inhibited the synthesis of endothelial cell DNA.