Effect of glycation on basic fibroblast growth factor induced angiogenesis and activation of associated signal transduction pathways in vascular endothelial cells: Possible relevance to wound healing in diabetes

摘要: Ineffectual wound healing in hyperglycaemic patients suffering from diabetes mellitus is characterised by a reduction capillary reformation (angiogenesis). Basic fibroblast growth factor (FGF-2) secreted fibroblasts, macrophages and particular endothelial cells (EC) response to tissue injury important promotion of neovascularisation. Recently, glycation FGF-2 has been shown significantly reduce its activity vitro. We have examined the kinetics compared ability with that native activate mitogenesis, formation associated signal transduction bovine aortic EC (BAEC). was exposed 0.25 M glucose-6-phosphate (G-6-P) for 24–72 h degree determined matrix assisted laser desorption ionisation mass spectrometry. Native heterogeneous Mw range 15,153.6–17,903 Da. After 24 incubation G-6-P there evidence glycation, increase corresponded addition 2.7 mol residues; after 48 h, 4 sugar added this increased 8.7 72 h. Dimerisation observed treatment. Induction mitogenesis BAEC reduced 25%–40% treatment 48–96 glycated (24 h) (gFGF-2;100 pg/ml–5 ng/ml; P < 0.05), whilst tubule between 60% 90% (100 pg/ml–1 0.05) 5 days FGF-2. Subsequent investigation molecules showed induced tyrosine phosphorylated proteins approximate 20–150 kDa 10 min particular, mitogen activated protein kinase (MAPK)/early (ERK-1, ERK-2), glycation. To determine reason angiogenic gFGF-2, we binding characteristics Total gFGF-2 cell surface analysed FACS (P 0.05). Further using 125I-labelled differentially washed samples, demonstrated significant high affinity receptor (46%) In summary, vitro occurs rapidly within presence elevated levels G-6-P. Glycation caused bind pathways responsible both BAEC. These results could help explain mechanism behind impaired mellitus.