p38 MAPK contributes to angiotensin II-induced COX-2 expression in aortic fibroblasts from normotensive and hypertensive rats

Objective To investigate the effect of angiotensin II on cyclooxygenase-2 (COX-2) expression in aortic adventitial fibroblasts from normotensive [Wistar-Kyoto (WKY)] rats and spontaneously hypertensive rats (SHRs). Methods Protein expression was determined by western blot, mRNA levels by real-time P...

Descripción completa

Detalles Bibliográficos
Autores: Beltran, AE, Briones, AM, Garcia-Redondo, AB, Rodriguez, C, Miguel, M, Alvarez, Y, Alonso, MJ, Martinez-Gonzalez, J, Salaices, M
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2009
País:España
Institución:Institut d’Investigació Biomèdica Sant Pau (IIB Sant Pau)
Repositorio:r-IIB SANT PAU. Repositorio Institucional de Producción Científica del Instituto de Investigación Biomédica Sant Pau
OAI Identifier:oai:iibsantpau.fundanetsuite.com:p13466
Acceso en línea:https://iibsantpau.fundanetsuite.com/Publicaciones/ProdCientif/PublicacionFrw.aspx?id=13466
http://hdl.handle.net/10115/3295
Access Level:acceso abierto
Palabra clave:angiotensin
cyclooxygenase-2
fibroblasts
hypertension
mitogen-activated protein kinase
Descripción
Sumario:Objective To investigate the effect of angiotensin II on cyclooxygenase-2 (COX-2) expression in aortic adventitial fibroblasts from normotensive [Wistar-Kyoto (WKY)] rats and spontaneously hypertensive rats (SHRs). Methods Protein expression was determined by western blot, mRNA levels by real-time PCR, transcriptional activity by luciferase assays, superoxide anion (O-2(center dot-)) production by dihydroethidine fluorescence and prostaglandin E-2 by enzyme immunoassay. Results Angiotensin II (0.1 mu mol/l, 0.5-6 h) time dependently induced COX-2 protein expression, this effect being transient in fibroblasts from WKY rats and maintained over time in SHRs. Angiotensin II effect was abolished by valsartan (1 mu mol/l), an angiotensin II type 1 receptor antagonist. Angiotensin II-induced prostaglandin E-2 production was reduced by valsartan and the COX-2 inhibitor NS398 (1 mu mol/l). Angiotensin II increased O-2(center dot-) production more in SHR than WKY rats. This increase was reduced by apocynin (30 mu mol/l) and allopurinol (10 mu mol/l), respective nicotinamide adenine dinucleotide phosphate (NADPH) and xanthine oxidase inhibitors. However, angiotensin II-induced COX-2 expression was unaffected by apocynin, allopurinol, tempol (1 mu mol/l) or catalase (1000 U/ml). Angiotensin II (2-30min) induced p38 mitogen-activated protein kinase (MAPK) phosphorylation, transiently in WKY rats but sustained in SHRs. The p38 inhibitor SB203580 (10 mu mol/l) reduced angiotensin II-induced COX-2 protein and mRNA levels. The angiotensin II effect was not prevented by inhibition of mRNA synthesis, and angiotensin II was unable to modulate COX-2 transcriptional activity. Conclusions Angiotensin II increases COX-2 expression in aortic fibroblasts through mechanisms including p38 MAPK pathway, independent of reactive oxygen species production and nonmediated by COX-2 transcriptional activity modulation. The sustained angiotensin-induced p38 MAPK activation in SHR cells might be related to the maintained COX-2 expression in this strain. J Hypertens 27: 142-154 (C) 2009 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.