CEFYBO   02669
CENTRO DE ESTUDIOS FARMACOLOGICOS Y BOTANICOS
Unidad Ejecutora - UE
artículos
Título:
Lipopolisaccharide plus interferon g treatment induces muscarinic acetylcholine receptors expression and function modulating NIH3T3 cell proliferation. Participation of nitric oxide synthase and cyclooxygenase.
Autor/es:
ESPAñOL ALEJANDRO; MADALENO, MARIANO; LOMBARDI, GABRIELA; CELLA, MAXIMILIANO; MARTINEZ PULIDO, PAOLA; SALES, MARIA ELENA
Revista:
BRITISH JOURNAL OF PHARMACOLOGY
Editorial:
WILEY-BLACKWELL PUBLISHING, INC
Referencias:
Lugar: Londres; Año: 2014 vol. 171 p. 5154 - 5167
ISSN:
0007-1188
Resumen:
Lypopolysaccharide (LPS) and interferon gamma (IFNã) are potent stimuli of inflammation, a process in which fibroblasts are frequently involved. We analyzed the effect of long-term treatment with LPS plus IFNg on the expression and function of muscarinic acetylcholine receptors (mAChR) in NIH3T3 fibroblasts regarding proliferation. We also investigated the participation of nitric oxide synthase (NOS) and cyclooxygenase (COX), and the role of the nuclear transcription factor kappa B (NF- kB) in this process. Experimental approach NIH3T3 cells were treated with LPS (10 ng.mL-1) plus IFNg (0.5 ng.mL-1) for 72 h (iNIH3T3 cells). Cell proliferation was evaluated with the MTT assay. Protein expression was analyzed by Western blot. NOS and COX activities were evaluated by Griess method and radioimmunoassay respectively. Key Results The cholinergic agonist carbachol was more effective to stimulate proliferation in iNIH3T3 than in NIH3T3 cells, probably due to the de novo induction of muscarinic acetylcholine receptor subtypes (M) M3 and M5, independently of NF-kB activation. iNIH3T3 cells produced higher amounts of nitric oxide (NO) and prostaglandin E2 (PGE2) than NIH3T3 cells, concomitantly with an up-regulation of NOS1 and COX-2, and with the de novo induction of NOS2 and NOS3 in inflamed cells. In addition, we found a positive feedback between NOS and COX in iNIH3T3 cells, which could potentiate inflammation. Conclusions and implications Inflammation induced muscarinic acetylcholine receptor expression and therefore carbachol-promoted proliferation. Inflammation also induced the expression of NOS isoenzymes and COX2, thus potentiating the action of carbachol on NO and PGE2 production. A positive cross-talk between NOS and COX triggered by carbachol in inflamed cells points to muscarinic acetylcholine receptors as therapeutic targets in inflammation.