Regulation Of MiRNA-150 Expression On The Recanalization Of Endothelial Progenitor Cells In Venous Thrombosis

Purpose: To determine the expression level of miR-150 in early endothelial cells(eEPCs)and late endothelial cells(lEPCs).To clarify the role of miR-150 biological functions in the differentiation,angiogenesis and so on for EPCs.To establish a rat model of athymic nude rats with venous thrombosis,and to investigate the effect of eEPCs and lEPCs alone and mixed transplantation on the treatment of thrombosis after adjusting the expression of miR-150.Methods: Mononuclear cells were isolated from peripheral blood by density gradient centrifugation,and EPCs were cultured in EGM-2MV medium.The morphological changes of EPCs were observed under inverted microscope.The flow cytometry,acetylated low density lipoprotein,and ulex europaeus agglutinin phagocytosis test were appliacated to identify the cells and to determine the expression content of miR-150 in the eEPCs and lEPCs.MiR-150 minics,inhibitors and controls were transfected into eEPCs and lEPCs by lentiviral vector.Fluorescence microscopy was used to analyze the efficiency of lentivirus transfection.The ability of new blood vessels after changing the expression of miR-150 in nude mice was detected by Matrigel gel.To establish a rat model of venous thrombosis in nude rats.And to transfer the eEPCs and lEPCs of the over expressed miR-150,which were mixed and mixed,into the model of venous thrombosis.After transplantion of 7d,the inferior vena cava thrombus segement were taken out of.The weighing and angipgraphy were performed to clear the thrombolytic recanalization in nferior vena cava thrombus segement.Results: The mononuclear cells obtained by centrifugation were cultured in EGM-2MV.The cells were cultured 2days in vitro and adhered to the wall after 2days.The adherent cells were fusiform,scattered in the cell colony formation.The cell colony gradually gather merged after 10-12 days.The shapes of cells showed cobblestone.At the same time,the corresponding antigen CD34,CD31,CD133,VEGFR-2,vWF,Dil-ac-LDL and FITC-UEA-1 double fluorescence uptake results showed double positive.Matigel tube formation test showede that the tubular and network like vascular structures were cultured in tissue.The eEPCs and the lEPCs were transfected by miR-150 minics,inhibitors and controls.It can be confirmed that the mi R-150 expression of surface antigen can regulate eEPCs and lEPCs.Up-regulation of miR-150 can promote the function of eEPCs.It also can accelerate thrombi resolution and recanalization.The expression levels of miR-150 in the eEPCs and lEPCs are different,and the regulation of miR-150 experssion can affect the ability of EPCs to promote the ability of thrombolysis and recanaliazationg.Conclusion: The content of miR-150 in the eEPCs and lEPCs were different.Regulation the expression of miR-150 can influence the function of EPCs,accelerate thrombolysis and promote recanalization in vessels.