Objectives To evaluate the effect of compound BA on angiogenesis in vitroand investigate its biological activity.
Methods Human Umbilical Vein Endothelial Cells (HUVECs) were treated by serious concentrations of BA for 24 h, 48 h, 72 h to explore the effect of compound BA on proliferation of HUVECs by MTT assay; HUVECs were planted on the matrigel, then treated by BA at concentrations of 1/2IC50, IC50, 2IC50 (IC50 is the concentration of BA at 50% inhibition rate). Take photos to record the tubes on matrigel during the tube formation process, identify the antiangiogenesis ability of compound BA by tube formation assay; in order to further explore its antiangiogenesis mechanism, we make HUVECs exposed to BA at concentraions of 1/2IC50, IC50, 2IC50 to investigate the effect of compound BA on expression of protein in signalling pathway PI3K/AKT.
Results Compound BA can inhibit proliferation of HUVECs, it is a dose-dependent manner during concentration of 2-8 μmol•L-1 (p < 0.05) and a time-dependent manner at 24 h, 48 h, 72 h (p < 0.05). Compound BA exhibits antiangiogenesis ability at concentration of 5.00 μmol•L-1 (IC50 value) (p < 0.05). While at concentration of 10 μmol•L-1 (2IC50 value), BA can totally kill all the HUVECs. Compared with the blank group, compound BA can not significantly inhibit tube formation at concentration of 2.5 μmol•L-1 (1/2IC50 value) (p > 0.05). The western Blotting result shows, compound BA can up-regulate the expression of protein PI3K, and down-regulate the expression of protein AKT, pAKT (Ser473) (P < 0.05), BA shows antiangiogenesis activity by surpressing the signal pathway PI3K/AKT.
Conclusions Sponge derived compound BA shows antiangiogensis activity invitro, its mechanism may be related to the signal pathway PI3K/AKT.