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 ce llular surface receptor (RAGE) induces translocation of the transcript ion factor NF-kappa B into the nucleus and NF-kappa B-mediated gene ex pression. This study examines the role of RAGE in the AGE albumin-medi ated induction of endothelial tissue factor, known to be partly contro lled by NF-kappa B. Methods and Results Endothelial cells (ECs) were i ncubated in the presence of an 18-mer phosphorothioate oligodeoxynucle otide antisense to the 5'-coding sequence of the RAGE gene (antisense RAGE; 0.1 mu mol/L). Sense oligonucleotides (sense RAGE, 0.1 mu mol/L) of the same region served as control. The cellular uptake of oligonuc leotides was controlled by immunofluorescence microscopy. RAGE transcr iption was suppressed by antisense RAGE, as demonstrated by RT-PCR rea ctions. AGE albumin-mediated activation of cultured ECs was studied af ter 48 hours of preincubation of ECs with antisense or sense RAGE. Ele ctrophoretic mobility shift assays and Western blot analysis demonstra ted that the AGE albumin-induced translocation of NF-kappa B 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.Conclusi ons Activation of ECs and induction of tissue factor by AGE albumin in ECs is dependent on RAGE.