Effects Of Endostatin And Pingyangmycin On Vascular Endothrlial Cell

Objective:To compare the effects of recombinant human endostatin (rHES) and pingyangmycin (PYM) using alone and in combination on human umbilical vein endothelial cell line (HUVEC cells), and discuss the relevant mechanisms, and reveal the rHES used in the treatment of proliferative hemangioma feasibility.Methods:Culture and subculture the HUVEC cells, to select the third to fifth generation of cells that grow well. After HUVEC cells were treated with different concentrations of rHES and PYM for 24h,48h, the inhibition rates on HUVEC cells were compared respectively by MTT assay, the changes before and after the drugs in cell morphology were observed under light microscope by Hematoxylin- eosin (HE) staining, the differences before and after the drugs of vascular endothelial cell growth factor (VEGF) in HUVEC cells were detected by immunohistochemistry and the apoptosis of HUVEC cells after treated by the drugs was analyzed by flow cytometry.Results:After were treated by rHES and PYM,the absorbance value (OD value) were decreased with the increase of drug concentration, the gray levels of the cells' cytoplasm were increased by VEGF immunohistochemical staining. The inhibition of HUVEC cells after the drugs show concentration dependence and time dependence. By light microscope, the morphological changes of the HUVEC cells that were treated with rHES were significant,which could be seen a large number of apoptotic cells that the number increased with increasing drug concentration, while after were treated with PYM the suspended cells increased and the number of adherent cells decreased largely with the increase of drug concentration. Apoptpsis of HUVEC cells which measured by flow cytometry was induced using by the two drugs alone, and showed the higher the concentration the more the number of apoptotic cells, the combination of the two drugs on HUVEC cells showed synergistic inhibitory effect.Conclusion:rHES and PYM both can inhibit effectively HUVEC cells, induce apoptosis and prevent VEGF into the cells to inhibit cell proliferation, indicating that the two drugs on inhibition of vascular endothelial cell mechanisms exist some common. Under the higher concentration of rHES apoptosis is still dominated, while under the concentration of PYM greater than a certain range (>20μg/ml) inhibition of the cells gradually led to necrosis. PYM on the impact of vascular endothelial cells adhesion was significantly higher than rHES, revealing that there are some different about two drugs on inhibition of vascular endothelial. rHES combined with PYM in inhibition of endothelial cells have synergistic effect, which provided the theoretical basis for the treatment of the proliferating hemangioma by the combination of the two drugs.