Influence and Significance of MMP-2 and VEGF of Hemangioma Endothelial Cells in Vitro Culture by Propranolol
|School||Zunyi Medical College,|
|Keywords||Hemangioma Vascular endothelial cells Cell culture Propranolol Vascular endothelial growth factor Matrix metalloproteinases-2|
Objective To observe the influence of VEGF and MMP-2 of hemangioma endothelial cells in vitro culture by propranolol. To study the mechanism of the hemangioma’s treatment by propranolol.Methods The hemangioma vascular endothelial cells which came from the specimens of infant hemangioma were cultured by explant combined with trypsin-digested technique.The expression of the factorⅧrelated antigen which only existed in endothelial cells to identify the hemangioma vascular endothelial cell were studied by the immunohistochemistry technique. When the cells were in the logarithmic phase,the culture medium was exchanged to serum-free medium to make the cell synchronization in 48 hours. Propranolol was used on cells at three different concentrationgs which were 0μmol/L,0.5μmol/L and 1.0μmol/L. Vascular endothelial growth factor(VEGF) and Matrix metalloproteinases-2(MMP-2) were detected by RT-PCR when the cells were cultured after 24 hours,while cell apoptosis were investigated by flow cytometry technique,and then the apoptosis related of VEGF and MMP-2 in relative mRNA expression level of vascular endothelial cells was analyzed.Results1.The hemangioma endothelial cells emigrated from the tissue began to grow fast and covered the bottom of culture bottles after three or four weeks. The expression factorⅦassociated antigen was positive in the endothelial cells in vitro culture.2. Compared with the apoptosis rate of hemangioma endothelial cells which were cultured after 24 hours, both of two propranolol treatment groups were significantly higher than the control group(P<0.01).The mRNA relative expression of VEGF was 1.02±0.42 in the group of 0.5μmol/L propranolol and 0.93±0.22 in the group of 1.0umol/L propranolol,both of them were lower than the control group which was 1.61±0.52(P<0.05). The relative expression of MMP-2 was 0.77±0.18 in the group of 0.5μmol/L propranolol and 0.67±0.27 in the group of 1.Oμmol/L propranolol,both of them were lower than the control group which was 1.47±0.57(P<0.05). Both of VEGF and MMP-2,compared with the 0.5μmol/L group, the difference was not statistically significant was found in 1.0μmol/L group(P>0.05).Conclusion Propranolol may promote the apoptosis or inhibit the growth of hemangioma vascular endothelial cells in vitro culture, and the mechanism might contribute to the reducing of mRNA expression of VEGF and MMP-2.