Redox regulation of ERK1/2 activation induced by sphingosine 1-phosphate in fibroblasts: Involvement of NADPH oxidase and platelet-derived growth factor receptor

• Published in: Biochimica et biophysica acta Vol. 1810; no. 4; pp. 446 - 456
• Main Authors: Catarzi, Serena, Romagnoli, Cecilia, Marcucci, Gemma, Favilli, Fabio, Iantomasi, Teresa, Vincenzini, Maria T
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2011
• Subjects: Animals; cell culture; Cell Proliferation; computer software; cytokines; enzyme-linked immunosorbent assay; fibroblasts; Fibroblasts - cytology; Fibroblasts - metabolism; fluorescence; hydrogen peroxide; Hydrogen Peroxide - metabolism; kinases; ligands; Lysophospholipids - metabolism; metabolites; Mice; mitogen-activated protein kinase; Mitogen-Activated Protein Kinase 1 - metabolism; Mitogen-Activated Protein Kinase 3 - metabolism; NAD(P)H oxidase (H2O2-forming); NADP (coenzyme); NADPH Oxidases - metabolism; NIH 3T3 Cells; oxidases; Oxidation-Reduction; platelet-derived growth factor; platelet-derived growth factor receptors; quantitative analysis; receptors; Receptors, Platelet-Derived Growth Factor - metabolism; signal transduction; spectral analysis; sphingolipids; sphingosine; Sphingosine - analogs & derivatives; Sphingosine - metabolism; transcriptional activation; tyrosine; Western blotting
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2011.01.005

BACKGROUND: Sphingosine 1-phosphate (S1P) is a sphingolipid metabolite synthesized after stimulation with growth factors or cytokines. S1P extracellular effects are mediated through specific Gi-protein coupled receptors (GPCRs). Recently, we demonstrated in NIH3T3 fibroblasts stimulated by platelet-derived growth factor (PDGF) or S1P the NADPH oxidase activation and the H₂O₂ intracellular level increase trough the Gi protein involvement. METHODS: NIH3T3 fibroblast cell cultures were used. Western blot and quantitative analyses by Chemidoc-Quantity-One software were performed. H₂O₂ level was assayed by fluorescence spectrophotometric analysis, and cell proliferation by counted manually or ELISA kit. RESULTS: This study demonstrates, in NIH 3T3 fibroblasts, a novel redox regulated mechanism of S1P-induced activation of ERK 1/2 related to NADPH oxidase activity and intracellular H₂O₂ level increase with PDGF receptor tyrosine kinase involvement through a transactivation mechanism. This event is mediated by S1P₁ and S1P₃ receptors by Gi proteins and can contribute to S1P mitogenic signaling. CONCLUSION: These results can be related to mechanisms of cross-talk previously identified between receptor tyrosine kinase, including PDGFreceptor, and several GPCR ligands. GENERAL SIGNIFICANCE: The redox-sensitive ERK1/2 and PDGFr tyrosine kinase activity could be targets for therapies in diseases in which deregulation of intracellular oxidative status and the consequent alteration of S1P and/or PDGF signaling pathway are involved.