蛇葡萄素的血管生成抑制作用

Inhibitory Effects of Ampelopsin on Angiogenesis

目的：探讨蛇葡萄素对内皮细胞及肿瘤细胞的生长及其血管生成的影响。方法：体外用MTT法对牛主动脉内皮细胞进行增殖抑制实验；用ELISA法、免疫组化染色法以及流式细胞仪观察蛇葡萄素对人肝璃细胞Bel-7402分泌VEGF、bFGF的抑制作用；体内观察蛇葡萄素对人肝癌Bel-7402裸鼠移植瘤的抑制作用。结果：蛇葡萄素对牛主动脉内皮细胞的增殖在6．4～51．2μg/ml呈现浓度依赖性抑制作用，IC50=22.0±4.0μg/ml。ELISA法测定结果显示，12．8μg/ml、25.6μg/ml、38．4μg/ml蛇葡萄素对肝癌Bel-7402细胞分泌VEGF的抑制率分别为14.2％、40．0％及49．6％。免疫组化染色结果显示，12．8μg/ml以卜的蛇葡葡素能使细胞体积缩小，分泌VEGF、hFGF明显减少。流式细胞仪检测VEGF、hFGF结果，25．6μg/ml及38．4μg/ml蛇葡萄索对VGEF的抑制率分别为32．2％及57.4％，对bFGF的抑制率分别为54．9％及62．6％。蛇葡葡素浓度为100、150、200mg／kg时对人肝痛Bel-7402裸鼠移植瘤的抑制率分别可达24．3％、41．4％及45．7％。结论：蛇葡萄素在体外能有效抑制内皮细胞增殖投人肝癌Bel-7402细胞分泌VEGF和bFGF，具有抑制血管牛成的作用；体内能有效抑制人肝癌Bel-7402裸鼠移植瘤的生长，提示蛇葡萄素也许可作为一个肿瘤血管牛成抑制剂用于肿瘤防治。

Objective： To study the effect of ampelopsin on angiogenesis. Methods：The anti-angiogenic effect was evaluated by MTr assay for proliferation of endothelial cells. The concentration of vascular endothelial growth factor （VEGF） and basic-fibroblast growth factor （bFGF） from human hepatocellular carcinoma Bel-7402 cells were detected by enzyme linked immunosorbant assay （ELISA）, Immunohistochemical staining was conducted to detect the expression of VEGF and hFGF. The VEGF and bFGF in the cancer cells were examined by flow cytometry. The inhibitory effect of ampelopsin on the growth of human hepatoeellular carcinoma Bel- 7402 in nude mice was studied. Results ： Ampelopsin was shown to inhit）it the proliferation of primary cultured bovine aortic endothelial cells in a concentration dependent manner in range of 6. 4 - 51.2 μg/ml. The IC50 （50% inhibilion concentration） value was 22.0 ± 4. 0 μg/ml. ELISA assay was shown that treatment with 12. 8 μg/ml,25, 6 μg/ml and 38.4 μg/ml of ampelopsin resulted in an inhibi- tion of VEGF production released by Be1-7402, and the inbibtitory rate was 14. 2% ,40. 0% and 49. 6% , respectively. After exposure to 12.8 μg/ml of ampelopsin, a decrease in the expression and activity of VEGF and bFGF was observed by immunohistochemical staining. The concentration of VEGF and bFGF secretion by Be1-7402 cells were lower following ampelopsin tream,ent as shown by flow cytometry. Treatment with 25.6μg/mL and 38.4 μg/ml of ampclopsin, the inbibilory rates were 32.2% and 57.4% for VEGF, and 54.9% and 62.6% for bFGF, respectively. The inhibitory rate of ampelupsin to the growth of the transplant tumor in nude mice were 24.3% ,41.4% and 45.75 respectively at the dose of 100 mg/kg,150 mg/kg and 200 mg/kg. Conclusion： Ampelopsin is a potent inhibitor of VEGF and bFGF expression and production in human hepatocellular earcinoma Bel-7402 cell, and may be a promising anglogenesis inhibitor.