Effects of AGEs on number and function of EPCs from human peripheral blood

Objective To study the effect of advanced glycation end products(AGEs) on number and biological characteristics of endothelial progenitor cells(EPCs) from cultured human peripheral blood. Methods Mononuclear cells(MNCs) isolated from human peripheral blood by density gradient centrifugation combined with adherent cell filtration were cultured. The biological functions of adherence cells were examined by the adsorption of Ulex europaeus agglutinin-1 (UEA-1) labeled by fluorescein isothiocyanate(FITC) and Dil-acLDL internalization on 4th day during culture. The expression of specific antigens(CD34, CD133) on cell surface was analyzed by cytometer. After 4 days,adherent cells were cultured with different concentrations of AGEs for 48 hours. The MTT assay and Boyden chamber were used to observe the proliferation and migration of EPCs,human fibronectin was used to examine adhesion ability of EPCs, in vitro angiogenesis assay kit was used to observe angiogenesis of EPCs which had been exposed to different concentrations of AGEs-HSA. Results AGEs dose-dependently decreased the number of EPCs (P 0. 01), weakened EPCs proliferation (P0. 01), migration (P0. 01), adhesive capacity (P 0. 05) ,and increased the apoptosis rate of EPCs in the early stage (P0. 01). Conclusion AGEs can decrease the number of EPCs from human peripheral blood and induce its dysfunction.