RESUMO
<p><b>OBJECTIVE</b>To observe the effect of platelet derived growth factor receptor β (PDGFR-β) transfected endothelial progenitor cells (EPCs) on vascular regeneration.</p><p><b>METHODS</b>Spleen-derived mononuclear cells (MNCs) were isolated using density gradient centrifugation and induced with special culture medium. EPCs transfection was performed with Lipofectamine(TM) 2000 reagent according to the instruction manual. Carotid artery injury was induced in splenectomized mice. EPCs were injected by tail vein immediately and at 24 h after endothelial injury of the carotid artery. Evans blue staining was performed to evaluate reendothelialization at 7 days after endothelial injury of the carotid artery.</p><p><b>RESULTS</b>Most adherent cells were LDL and UEA-I double positive. Laser scanning confocal microscopy showed that transfection efficiency was about 50%-60%. The reendothelialized area in the PDGFR-β-EPCs group was significantly larger than that in EGFP-EPCs group.</p><p><b>CONCLUSION</b>Transplantation of PDGFR-β over-expressed EPCs can promote reendothelialization in the early phase after carotid artery injury.</p>
Assuntos
Animais , Masculino , Camundongos , Lesões das Artérias Carótidas , Patologia , Cirurgia Geral , Células Cultivadas , Modelos Animais de Doenças , Células Endoteliais , Biologia Celular , Metabolismo , Transplante , Endotélio Vascular , Biologia Celular , Camundongos Endogâmicos C57BL , Receptores do Fator de Crescimento Derivado de Plaquetas , Genética , Metabolismo , Regeneração , Transplante de Células-Tronco , Células-Tronco , Biologia Celular , MetabolismoRESUMO
Objective To investigate the different influences of simvastatin on proliferation of rat smooth muscle progenitor cells(SPCs) versus endothelial progenitor cells (EPCs) and identify the compounds that differentially inhibit SPCs and EPCs proliferation for clinical usefulness. Methods Total mononuclear cells (MNCs) were isolated from bone marrow of rats by Fieoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. SPCs outgrew from the culture of MNCs in the presence of platelet-derived growth factor-BB and basic fibroblast growth factor, whereas EPCs were obtained in the presence of vascular endothelial growth factor. SPCs were identified as adherent cells positive for α-smooth muscle actin (α-SMA) by indirect immunofluoreseent staining. EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining. SPCs and EPCs were stimulated by simvastatin (0.01～10.00 μmol/L) or vehicle control for the respective time points (6 h, 12 h, 24 h and 48 h). SPCs and EPCs proliferation were assayed with 3H-TdR incorporation and manual counting respectively. Results Simvastatin obviously inhibited SPCs proliferation. At the concentration of 0. 01 μmol/L for 12 h,simvastatin significantly reduced the number of SPCs by (5.8±3.1)% compared with control group (P<0.05). Simvastatin significantly stimulated EPCs proliferation, which was dose- and time dependent and reached maximum at 1 μmol/L after 24 hours (2.0±0.1 fold increase, P<0.01).Conclusions Simvastatin displays different effects on SPCs (inhibited) and EPCs (promoted)proliferation. Local application of simvastatin may inhibit arterial restenosis and promote reendothelialization of injured vessels.