Propranolol For Haemangiomas-insight Into VEGF/ERK Signaling Transduction In HUVEC-12 Cells

Objectives1. To investigate the influence of Propranolol on the the cytotoxicity and proliferation of the endothelial cells in vitro.2. To investigate the action of propranolol involved haemangiomas related to VEGF/ERK signaling transduction (ERK:extracellular signal related kinases) in cultured human umbilical vein endothelial-12 cells (HUVEC-12 cells)Methods1. Endothelial cells were treated with different dose of propranolol, the cell morphologcal changes were observed by light microscopy; the cell proliferation activity and cell viability was detected by MTT assay; p-ERK1/2 expressions were assessed by western bloting.2. Endothelial cells were treated with different dose of propranolol, p-ERK1/2 expressions were assessed by western blot.3. Endothelial cells induced by VEGF were treated with different dose of propranolol, the cell proliferation activity and cell viability was detected by MTT assay, p-ERK1/2 expressions were assessed by western bloting.4.In the study of intervention, endothelial cells induced or not by VEGF were treated by PD98059 (specific inhibitor of ERK) and/or propranolol, the cell proliferation activity and cell viability was detected by MTT assay, p-ERK1/2 expressions were assessed by western bloting.Results1. propranolol can slight activated proliferation of endothelial cells(P<0.05) and it has no impacts on expression of p-ERK1/2 and cell viability(P>0.05).2. The VEGF positive system of endothelial cells was estabished when VEGF is of 20ng/ml,in which the p-ERK1/2 is highest expression(P<0.05).3.The proliferation and the expression of p-ERK1/2 of endothelial cells induced by VEGF were inhibited by propranolol at the concentration between 8-16 ug/ml (P<0.05),and cell viability have no significant changes(P>0.05)4. Cell viability and proliferation have no significant changes when PD98059 at the concentrtion of 12.5umol/l,and PD98059 at this concentration can inhibited the expression of p-ERK1/2 and cell proliferation induced by VEGF.And we also find that propranolol has the same effect as PD98059,and its efficacy may be of synergy.ConclusionThe mechanism of propranalol control the growth of infantile haemangiomas may be due to inhibite the proliferation of EC induced by VEGF, for it has specific negative effect on ERK pathway.