Angiotensin II activates nuclear transcription factor κB through AT1 and AT2 in vascular smooth muscle cells -: Molecular mechanisms

Nuclear factor-kappa B (NF-kappa B) regulates many genes involved in vascular physiopathology. We have previously observed in vivo NF-kappa B activation in injured vessels that diminished by angiotensin-converting enzyme inhibition. In the present work, we investigated the effect of angiotensin II (Ang II) on NF-kappa B activity in rat vascular smooth muscle cells, evaluating the molecular mechanisms and the specific receptor subtype involved. Ang II increased NF-kappa B DNA binding (5-fold, 10(-9) mol/L at 1 hour; electrophoretic mobility shift assay), nuclear translocation of p50/p65 subunits, and cytosolic inhibitor kappa B alpha (I kappa B alpha) degradation. Ang II elicited NF-kappa B-mediatcd transcription (transfection of a reporter gone) and expression of NF-kappa B-related genes (monocyte chemoattractant protein-1 and angiotensinogen). AT(1) (DUP753) and AT(2) (PD123319 and CGP42112) receptor antagonists inhibited Ang II-induced NF-kappa B DNA binding in a dose-dependent manner (approximate to 85% for each one; 10(-5) mol/L at I hour). The AT(2) agonist p-aminophenylalanine(6)-Ang II augmented NF-kappa B binding (4.6-fold, 10(-9) mol/L at 1 hour), p65 nuclear levels, and transcription of an NF-kappa B reporter gene. AT(1) antagonist markedly inhibited NF-kappa B-mediated transcription and gene expression. Some differences between AT(1)/AT(2) intracellular signals were found. Antioxidants and ceramide inhibitors, but not protein kinase C inhibitors, diminished NF-kappa B activation elicited by both Ang II and the AT(2) agonist, while tyrosine kinase inhibitors only decreased Ang II-induced NF-kappa B activity. Our results demonstrate that Ang II activates NF-kappa B via AT(2) and AT(2), although NF-kappa B-mediated transcription occurred mainly through AT(1). Both receptors share some signaling pathways (oxygen radicals and ceramide); however, tyrosine kinases only participate in AT(1)/NF-kappa B responses. These data provide novel insights into Ang II actions, suggesting a potential implication of the AT(2) in the pathobiology of vascular cells.