c-Jun NH2-Terminal Kinase Inhibitor Anthra(1,9-cd)pyrazol-6(2H)-one Reduces Inducible Nitric-Oxide Synthase Expression by Destabilizing mRNA in Activated Macrophages
Open Access
- 1 August 2003
- journal article
- Published by Elsevier in Molecular Pharmacology
- Vol. 64 (2) , 308-315
- https://doi.org/10.1124/mol.64.2.308
Abstract
In this study, we investigated the role of c-Jun NH2-terminal kinase (JNK), a member of the mitogen-activated protein kinase (MAPK) family, in lipopolysaccharide (LPS)-stimulated inducible nitric-oxide synthase (iNOS) expression and nitric oxide (NO) production in J774 murine macrophages. Anthra(1,9-cd)pyrazol-6(2H)-one (SP600125), a pharmacological inhibitor of JNK, inhibited phosphorylation of c-Jun with an IC50 of 5 to 10 μM. At the same concentrations, SP600125 inhibited LPS-induced iNOS protein expression and NO production. SP600125 had no effect on the activation of nuclear factor κB, which is an important transcription factor for iNOS expression. SP600125 had no significant effect on iNOS mRNA levels if measured 4 h after LPS. In contrast, SP600125 reduced iNOS mRNA levels >90% when measured 8 h after LPS. These data suggest that SP600125 reduced iNOS mRNA stability, and this was confirmed in the mRNA degradation assay using actinomycin D, in which SP600125 reduced the iNOS mRNA half-life from 5 to 2 h. These results show that the JNK pathway is involved in the up-regulation of LPS-induced iNOS expression and NO production by a mechanism related to the stabilization of iNOS mRNA.Keywords
This publication has 28 references indexed in Scilit:
- MAPKK-Independent Activation of p38α Mediated by TAB1-Dependent Autophosphorylation of p38αScience, 2002
- Induction of Inducible Nitric Oxide Synthase-NO· by Lipoarabinomannan ofMycobacterium tuberculosisIs Mediated by MEK1-ERK, MKK7-JNK, and NF-κB Signaling PathwaysInfection and Immunity, 2001
- SB 203580 is a specific inhibitor of a MAP kinase homologue which is stimulated by cellular stresses and interleukin‐1Published by Wiley ,2000
- Interleukin-1β-induced Rat Pancreatic Islet Nitric Oxide Synthesis Requires Both the p38 and Extracellular Signal-regulated Kinase 1/2 Mitogen-activated Protein KinasesJournal of Biological Chemistry, 1998
- Upstream NF-κB Site Is Required for the Maximal Expression of Mouse Inducible Nitric Oxide Synthase Gene in Interferon-γ plus Lipopolysaccharide-Induced RAW 264.7 MacrophagesBiochemical and Biophysical Research Communications, 1997
- NITRIC OXIDE AND MACROPHAGE FUNCTIONAnnual Review of Immunology, 1997
- Nitric Oxide Synthase Type II mRNA Stability Is Translation‐ and Transcription‐DependentJournal of Neurochemistry, 1996
- PD 098059 Is a Specific Inhibitor of the Activation of Mitogen-activated Protein Kinase Kinase in Vitro and in VivoJournal of Biological Chemistry, 1995
- JNK1: A protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domainPublished by Elsevier ,1994
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976