SiRNA-mediated Silencing Of PAI-1 Gene Acts As A Promoter Over The Recanalization Of Endothelial Progenitor Cells In Venous Thrombosis

With the changing lifestyle,venous thrombosis(VT)is becoming increasingly prevalent and poses a burden on the health economy.With the rise of stem cell therapy,more and more studies have focused on the role of endothelial progenitor cells(EPCs)in thrombosis recanalization and its potential clinical value.EPCs are multipotent progenitor cells in bone marrow,which are recruited to VT lesion sites under the pathological conditions of thrombosis and play a role in thrombolysis by repairing damaged blood vessels.With the in-depth study of EPCs reinfusion therapy,more and more studies began to focus on the exploration of combined treatment strategies of EPCs.Plasminogen activator inhibitor 1(PAI-1)is a major negative regulator of plasmin-driven protein hydrolysis in fibrinolytic system.PAI-1 is a potential target for VT therapy by inhibiting the activation of plasminogen fibrinolytic enzymes by inhibiting the effects of tPA and uPA during fibrinolysis.At present,the role of PAI-1 in EPCs-mediated thrombosis has not been fully elucidated.We aimed to investigate the effect of plasminogen activator inhibitor 1(PAI-1)silencing on the recanalization of VT in rat EPCs.EPCs and VT rat models were cultured and treated with negative control siRNA vector and PAI-1-siRNA vector,respectively.4,5-Dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide assay,wound-healing test,and Matrigel-induced tubular experiment were performed to detect the ability of cell proliferation,migration,and EPCs lumen formation.Immunohistochemistry was used to observe the recanalization of thrombus.The messenger RNA(mRNA)and protein expression of PAI-1 and vascular endothelial growth factor(VEGF)were determined by everse transcription quantitative polymerase chain reaction and Western blot analysis.PAI-1-siRNA enhances the luminal formation ability of EPCs and significantly promotes EPCs homing.In response to PAI-1 gene silencing,tissues from inferior vena cava displayed reduced mRNA and protein expression of PAI-1,increased VEGF expression as well as promoted lumen-like structures.PAI-1 gene silencing can promote the recanalization of VT by enhancement of the luminal formation ability of rats' EPCs.