Objective   To investigate the effects of glycated fibroblast growth factor-2 (gFGF-2) on the number and the biological activity of endothelial progenitor cells (EPCs) from rat bone marrow in vitro.   Methods   Recombinant FGF2 at the dose of 10 μg/ml was incubated with fructose at the dose of 250 mmol/L at 37 ℃ for 48 h. Glycated proteins were analyzed by matrix-assisted laser desorption/ionisation mass spectrometry (MALDI). Total mononuclear cells (MNCs) were isolated from bone marrow by Ficoll density gradient centrifugation. After cultured for 4 d

MNCs were divided into three groups: the first group cultivated in FGF-2 (150 μg/ml)

the second group cultivated in gFGF-2 (150 μg/ml)

and the blank group cultivated without any growth factor for another 3 d×respectively. The shape/growth/multiplication conditions were observed by inverted phase contrast microscopy. EPCs were identified using CD34

CD133

VEGFR-2

ac-LDL

and UEA-1 by immunohistochemical and immunofluorescent stainings. The attached cells were collect at 7 d after culture by trypsinization. Then cells were incubated at 37 ℃ for 72 h with gFGF-2 (or FGF-2). The angiogenic activity was determined with an in vitro angiogenesis kit. The attachment ability of the cells at 7 d after culture was detected. The migration ability of EPCs was determined with the modified Boyden cave method. The reproductive activity was determined by MTT method. The amount of EPO

GM-CSF and IL-8 in cell culture solution was also determined by radioactivity immunization.   Results   Protein fingerprint analysis showed that FGF-2 was successfully glycated to gFGF-2. Test of angiogenic capability in gFGF-2 group and FGF-2 group in vitro indicated that glycated FGF-2 could inhibit angiogenic activity (P0.05). The number

adhesion ability

migration ability

reproductive activity

EPCs secreted amount of EPO

GM-CSF

and GM-IL-8 in gFGF-2 group were lower than those in FGF-2 (P0.05).   Conclusion   The decreased number

the lowered biological activity

and weakened angiogenic ability of EPCs originated from rat bone barrow in vitro after culture with gFGF-2 may suggest that gFGF-2 may be one of the factors leading to impairment of angiogenesis in diabetes mellitus patients.