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Aim:To investigate whether aspirin has an influence on endothelial progenitorcells(EPC).Methods:Total mononuclear cells(MNC)were isolated from periph-eral blood by Ficoll density gradient centrifugation,then cells were plated onfibronectin-coated culture dishes.After 7 d of culture,attached cells were stimu-lated with aspirin(to achieve final concentrations of 1,2,5,and 10 retool/L)for 3,6,12,and 24 h.EPC were characterized as adherent cells that were double positivefor 1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine low density lipoprotein(DiLDL)uptake and lectin binding by direct fluorescent staining.EPC prolifera-tion and migration were assayed using a 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide(MTT)assay and a modified Boyden chamber assay,respectively.An EPC adhesion assay was performed by replating the EPC onfibronectin-coated dishes,and then adherent cells were counted.In vitrovasculogenesis activity was assayed by using an in vitro vasculogenesis kit.Inducible nitric oxide synthase(iNOS)was assayed by Western blotting.Results:Incubation of isolated human MNC with aspirin decreased the number of EPC.Aspirin also decreased the proliferative,migratory,adhesive,and in vitrovasculogenesis capacity of EPC,and also their iNOS levels in a concentration-and time-dependent manner.Conclusion:Aspirin decreases(1)the number ofEPC;(2)the proliferative,migratory,adhesive and in vitro vasculogenesis capaci-ties of EPC;and(3)iNOS levels in EPC.
Aim: To investigate whether aspirin has an influence on endothelial progenitor cells (EPC). Methods: Total mononuclear cells (MNC) were isolated from periphral blood by Ficoll density gradient centrifugation, then cells were plated on fibronectin-coated culture dishes. After 7 d of culture, attached cells were stimu-lated with aspirin (to achieve final concentrations of 1,2,5, and 10 retool / L) for 3,6,12, and 24 h. EPCs were characterized as adherent cells that were double positivefor 1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine low density lipoprotein (DiLDL) uptake and lectin binding by direct fluorescent staining. EPC prolifera tion and migration were assayed using a 3- (4,5-dimethyl-2 thiazoyl) -2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and a modified Boyden chamber assay, respectively. An EPC adhesion assay was performed by replating the EPC on fibronectin-coated dishes, and then adherent cells were counted. In vitro vasculogenesis activity was assayed by using an in vitro vasculogenesis kit. Ind Results: Incubation of isolated human MNC with aspirin decreased the number of EPC. Aspirin also decreased the proliferative, migratory, adhesive, and in vitro vasculogenesis capacity of EPC, and also their iNOS levels (iNOS) was assayed by Western blotting (2) the proliferative, migratory, adhesive and in vitro vasculogenesis capaci-ties of EPC; and (3) iNOS levels in EPC.