Objective: To study the effect of ampelopsin on angiogenesis.
Methods: The anti-angiogenic effect was evaluated by MTT 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 bFGF. The VEGF and bFGF in the cancer cells were examined by flow cytometry. The inhibitory effect of ampelopsin on the growth of human hepatocellular carcinoma Bel-7402 in nude mice was studied.
Results: Ampelopsin was shown to inhibit the proliferation of primary cultured bovine aortic endothelial cells in a concentration dependent manner in range of 6.4 - 51.2 microg/ml. The IC50 (50% inhibition concentration) value was 22.0 +/- 4.0 microg/ml. ELISA assay was shown that treatment with 12.8 microl/m1, 25.6 microl/ml and 38.4 microg/ml of ampelopsin resulted in an inhibition of VEGF production released by Bel-7402, and the inbibtitory rate was 14.2%, 40.0% and 49.6%, respectively. After exposure to 12.8 microg/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 Bel-7402 cells were lower following ampelopsin treatment as shown by flow cytometry. Treatment with 25.6 microg/mL and 38.4 microg/ml of ampelopsin, the inbibitory rates were 32.2% and 57.4% for VEGF, and 54.9% and 62.6% for bFGF, respectively. The inhibitory rate of ampelopsin 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 carcinoma Bel-7402 cell, and may be a promising angiogenesis inhibitor.