葡萄抑制新生血管形成的实验研究

The experimental study on antiangiogenesis of four grape varieties

目的探讨四种不同品系葡萄提取物对人脐静脉内皮细胞(HUVEC)形成新生血管及分泌基质金属蛋白酶-2(MMP-2)的抑制作用。方法新鲜葡萄丙酮法提取,低温保存;葡萄提取物与血管内皮细胞在Matrigel(Ⅳ型胶原)上混合培养,形态学观察提取物抑制血管形成能力;葡萄提取物与血管内皮细胞在37℃,5%CO2条件下混合培养,6小时后取培养液在含有Gelatin(明胶蛋白)的凝胶上进行电泳(酶谱分析)检测葡萄提取物对MMP-2的抑制能力。结果四种葡萄均具有较强的抑制新生血管形成的能力,在5 mg/ml时均可部分抑制HUVEC在Matrigel上形成新生血管,20 mg/ml时可以完全抑制新生血管的形成。除Chardonnay外,其余三种葡萄在20 mg/ml均可完全抑制HUVEC的MMP-2的分泌。抑制新生血管形成的能力与植物化学物质(phytochemicals)的含量呈正相关。结论葡萄提取物抑制新生血管形成主要归功于其所含的植物化学物质,富含植物化学物质的葡萄具有某种程度的抗癌功效。

Objective To investigate the inhibitory effects of grape on human umbilical vein endothelial cells （HUVEC） capillary tube formation and matix - metalloproteinase - 2 （MMP- 2） expression secreted into the medium. Methods Four different grape varieties （Concord, Niagara, Chardonnay and Pinot Noir） were extracted using acetone and stored at - 70℃ ; the total amount of phenolics and flavonoids for each of the four grape varieties were determined by spectrophotometry; the grape extracts were co - cultured with HUVEC on Matrigel and effects of inhibiton on tube formation were obselved by microscope, inhibited effects of grape extracts on MMP- 2 expression were examined by zymogram. Results All four grape varieties inhibited the tube formation of HUVEC with a concentration - dependent manner on Matrigel and almost no tube formation at 20 mg/ml of grape extracts. Except Chardonnay, the other three grape varieties inhibited secretion of MMP - 2 completely at 20 mg/ml. There is a positive relationship between the total phenolics/flavonoids and antiangiogenical activities. Conclusion The grapes tested have the potential to irdfihit angiogenesis mainly by their phenolics/flavonoids content, which partly contribute to their cancer chempreventive efficacy.