Interaction of Nε(carboxymethyl)lysine- and methylglyoxal-modified albumin with endothelial cells and macrophages. Splice variants of RAGE may limit the responsiveness of human endothelial cells to AGEs
article
In diabetes mellitus an increased risk exists for vascular complications. A role for advanced glycation endproducts (AGEs) in the acceleration of vascular disease has been suggested. Nε-(carboxymethyl)lysine (CML)- and methylglyoxal (MGO)-modified proteins have been identified as major AGEs. The interaction of these AGEs with the human enclothelial cells and macrophages was studied. Changes in adhesion molecule expression, i.e. vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1) and E-selectin were determined by cell-bound Elisa on human endothelial cells after incubation with CML-modified albumin and MGO-modified albumin. The presence of the full-length receptor of AGEs (RAGE) and splice variants of RAGE was determined by specific RT-PCR. In addition, binding studies were performed with CML- and MGO-modified albumin to endothelial cells and P388DI macrophages. We demonstrated that CML-albumin or MGO-albumin did not induce activation of endothelial cells as measured by the expression of adhesion molecules, while, under the same conditions, TNF-α did. No specific binding of CML-albumin and MGO-albumin on these was found. In contrast to endothelial cells, a specific binding of MGO-albumin to P388DI macrophages was demonstrated, which could be competed by ligands of scavenger receptors. In human umbilical vein and microvascular endothelial cells we found the N-truncated and C-truncated splice variants of RAGE. In conclusion, under our experimental conditions no CML- or MGO-albumin-induced increase in adhesion molecule expression was found on endothelial cells. In agreement with this, no binding of these AGEs was found to endothelial cells. The existence of splice variants of RAGE in endothelial cells might explain the lack of interaction of extracellular AGEs with these cells. © 2006 Schattauer GmbH, Stuttgart. Chemicals / CAS: advanced glycation end product receptor, 198785-73-8, 247590-69-8; endothelial leukocyte adhesion molecule 1, 128875-25-2; intercellular adhesion molecule 1, 126547-89-5; methylglyoxal, 78-98-8; lysine, 56-87-1, 6899-06-5, 70-54-2; serum albumin, 9048-46-8; advanced glycosylation end-product receptor; Cell Adhesion Molecules; Glycosylation End Products, Advanced; Lysine, 56-87-1; Protein Isoforms; Pyruvaldehyde, 78-98-8; Receptors, Immunologic; Serum Albumin
Topics
Biomedical ResearchAdhesion moleculesCarboxymethyllysineEndothelial activationHuman microvascular endothelial cellsHUVECsMethylglyoxal6 n carboxymethyllysineadvanced glycation end productadvanced glycation end product receptoralbumincell adhesion moleculeendothelial leukocyte adhesion molecule 1intercellular adhesion molecule 1ligandscavenger receptortumor necrosis factor alphavascular cell adhesion molecule 1advanced glycosylation end product receptoradvanced glycosylation end-product receptordrug derivativeimmunoglobulin receptorisoproteinbinding competitioncell activationcontrolled studyendothelium cellenzyme linked immunosorbent assayhuman cellmicrovasculaturemolecular interactionnucleotide sequenceprotein bindingprotein expressionprotein modificationprotein variantreverse transcription polymerase chain reactionRNA splicingumbilical veinvascular endotheliumalternative RNA splicingchemistrycytologygeneticsmetabolismAlternative SplicingCell Adhesion MoleculesEndothelial CellsEndothelium, VascularGlycosylation End Products, AdvancedHumansLysineMacrophagesProtein IsoformsPyruvaldehydeReceptors, ImmunologicSerum Albumin
TNO Identifier
239112
ISSN
03406245
Source
Thrombosis and Haemostasis, 95(2), pp. 320-328.
Pages
320-328
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