Advanced glycation end product (AGE)-mediated induction of tissue factor in cultured endothelial cells is dependent on RAGE.
BACKGROUND
Binding of advanced glycation end products (AGEs) to the cellular surface receptor (RAGE) induces translocation of the transcription factor NF-kappaB into the nucleus and NF-kappaB-mediated gene expression. This study examines the role of RAGE in the AGE albumin-mediated induction of endothelial tissue factor, known to be partly controlled by NF-kappaB.


RESULTS
Endothelial cells (ECs) were incubated in the presence of an 18-mer phosphorothioate oligodeoxynucleotide antisense to the 5'-coding sequence of the RAGE gene (antisense RAGE; 0.1 micromol/L). Sense oligonucleotides (sense RAGE, 0.1 micromol/L) of the same region served as control. The cellular uptake of oligonucleotides was controlled by immunofluorescence microscopy. RAGE transcription was suppressed by antisense RAGE, as demonstrated by RT-PCR reactions. AGE albumin-mediated activation of cultured ECs was studied after 48 hours of preincubation of ECs with antisense or sense RAGE. Electrophoretic mobility shift assays and Western blot analysis demonstrated that the AGE albumin-induced translocation of NF-kappaB from the cytoplasm into the nucleus was suppressed in the presence of antisense RAGE but not by sense RAGE. In parallel, AGE albumin-mediated tissue factor transcription, activity, and antigen were significantly reduced in ECs exposed to antisense RAGE, whereas sense RAGE (and nonspecific oligonucleotides) did not influence tissue factor expression.


CONCLUSIONS
Activation of ECs and induction of tissue factor by AGE albumin in ECs is dependent on RAGE.
Bierhaus
A
A
Department of Internal Medicine I, University of Heidelberg, Germany.
Illmer
T
T
Kasper
M
M
Luther
T
T
Quehenberger
P
P
Tritschler
H
H
Wahl
P
P
Ziegler
R
R
Müller
M
M
Nawroth
P P
PP
D000818:Animals; D001011:Aorta; D002417:Cattle; D002478:Cells, Cultured; D015536:Down-Regulation; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D017127:Glycation End Products, Advanced / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000096:biosynthesis* / Q000502:physiology; D012710:Serum Albumin, Bovine / Q000378:metabolism / Q000494:pharmacology*; D013925:Thromboplastin / Q000096:biosynthesis*; D014158:Transcription, Genetic / Q000187:drug effects; D014471:Umbilical Veins
D017127:Glycation End Products, Advanced; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; C081628:advanced glycation end products-bovine serum albumin; D012710:Serum Albumin, Bovine; D013925:Thromboplastin