The Study On The Treatment Of Transplantation Of Vascular Endothelial Progenitor Cells With The Modification Gene Of VEGF165on The Deep Vein Withendothelium Injury After Vein Thrombectomy

Objective：To explore the therapeutic effect of vascular Endothelial Progenitor cellswith the modification gene of VEGFl65in the deep vein with post-thrombosis syndrome.Methods：1The bone marrow mononuclear cells derived from Sprague-Dawley ratswere isolated by Ficoli density gradient centrifugation and cultured by EBM-2medium，．2The adenoviruses being amplified by human embryo renal cells(HEK293A) were used totransfect EPCs for higher efficacy．3Rat thrombosis models(weight of some300g) wereestablished by performing external banding valvuloplasty between the proximal end of therenal veins and the proximal part of lumbar veins of inferior vena cava and ferric chloridewas applied to the external vein surface.The rats were divided into four groups randomly，and12animals in each group．4Vein thrombectomy is operated3～5days later and theEPCs injected into inferior vena cava. Group A：Ad—VEGF165transfection group，lml×10~6/mlAd—VEGF165一EPCs transplantation； Group B：Ad-GFP transfectionGroup，1ml×10~6/ml Ad．GFP．EPCs transplantation； Group C: EPCs group，lml×10~6/ml EPCs transplantation； Group D：control group，1ml of medium transplantation．5Collecting specimens of the thrombus section of vena cava inferior at different time．HEstaining and vWF immunohistochemistry staining including The antibodies of beta-actin、VEGF and MMP-2protein expression levels are used to observe the organisation of newthrombus and the repair of the Tunica intima of vein; a scanning electron microscopy isused to observe the Tunica intima of vein status.Results:(1) Bone marrow mononuclear cells can be induced to differentiate to generate EPCs and the late Endothelial Progenitor cells can be obtained during thesubculture of the cells subsequencely;(2)As the carrier, high efficiency of HEK293A cellscan amplify adenovirus for a high titer and transfected EPCs more efficiently;(3)It’sefficient and convenient for using ferric chloride to build rat models of the inferior venacava thrombosis model:(4) Gene-modified EPCs transplantation can promote thepost-thrombotic Tunica intima of vein repair significantly.Conclusion: The transplantation of EPCs transfected by VEGFl65gene can amenddeep vein wall through promoting Tunica intima of vein’s repair significantly and enhancethe efficacy of. vein thrombectomy.