Effect Of Exogenous Spermine On Angiogenesis Of Porcine Umbilical Vein Endothelial Cells And Its Mechanism

The density and size of the placental blood vessels determine the perfect function of the placenta with great significance for affecting the performance of the sow.Studying the formation of placental blood vessels is of great value in improving the performance of the sows and the healthy growth of the embryos.Spermine is an aliphatic amine synthesized by cells and widely exists in mammals,being significant for cell proliferation and differentiation.This project aimed to study the influences of exogenous spermine on expression of angiogenesis-related gene and protein and from endothelial cells of porcine umbilical vein by applying the techniques of cell biology at the molecular and cellular levels,providing theoretical support for seeking method to prevent intrauterine growth retardation of piglets derived from incomplete function of the sow's placenta.To this end,the projects were carried out as follows:1.The umbilical cord of normal parturition pig was as material.The umbilical vein was perfused with type I collagenase before digesting at 37 ? for 15 min,which had better separation effect of endothelial cells.High concentration of serum was applied to promote the growth of primary cells.The endothelial cells were identified by morphology evidence and staining of anti-CD31 protein,anti-vWF protein after separation and purification.2.Different concentrations(0?M,2.5?M,5?M,7.5?M,10?M)of spermine affected the proliferation of porcine umbilical vein endothelial cells to varying degrees.The addition of 7.5?M spermine greatest affected on the proliferation of endothelial cells(P<0.05).Therefore cell migration and lumen formation assays were performed using the addition of 7.5?M spermine as the test group.The number of cell migration and lumen formation in the treatment group of 7.5?M spermine addition were both significantly higher than those in control group(P<0.05).Thus the addition of 7.5?M spermine promoted angiogenesis of umbilical vein endothelial cells in vitro.3.The gene expression levels of FGF-2,PLGF,ANG-1,VEGF-A and PI3 K,AKT1,m-TOR,S6K1,MEK were detected in control group and treatment group of 7.5?M spermine treatment by real-time PCR.The results of real-time PCR showed that the gene expression levels of FGF-2,PLGF,m-TOR and MEK in 7.5?M spermine treatment group were significantly higher than those in control group(P<0.05).