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Am J Physiol Cell Physiol 280: C441-C450, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 3, C441-C450, March 2001

IFN-gamma  + LPS induction of iNOS is modulated by ERK, JNK/SAPK, and p38mapk in a mouse macrophage cell line

Edward D. Chan1 and David W. H. Riches2

1 Division of Pulmonary Sciences and Critical Care Medicine, University of Colorado Health Sciences Center, and 2 Program in Cell Biology, National Jewish Medical and Research Center, Denver, Colorado 80206

Nitric oxide (NO·) produced by inducible nitric oxide synthase (iNOS) mediates a number of important physiological and pathophysiological processes. The objective of this investigation was to examine the role of mitogen-activated protein kinases (MAPKs) in the regulation of iNOS and NO· by interferon-gamma (IFN-gamma ) + lipopolysaccharide (LPS) in macrophages using specific inhibitors and dominant inhibitory mutant proteins of the MAPK pathways. The signaling pathway utilized by IFN-gamma in iNOS induction is well elucidated. To study signaling pathways that are restricted to the LPS-signaling arm, we used a subclone of the parental RAW 264.7 cell line that is unresponsive to IFN-gamma alone with respect to iNOS induction. In this RAW 264.7gamma NO(-) subclone, IFN-gamma and LPS are nevertheless required for synergistic activation of the iNOS promoter. We found that extracellular signal-regulated kinase (ERK) augmented and p38mapk inhibited IFN-gamma  + LPS induction of iNOS. Dominant-negative MAPK kinase-4 inhibited iNOS promoter activation by IFN-gamma  + LPS, also implicating the c-Jun NH2-terminal kinase (JNK) pathway in mediating iNOS induction. Inhibition of the ERK pathway markedly reduced IFN-gamma  + LPS-induced tumor necrosis factor-alpha protein expression, providing a possible mechanism by which ERK augments iNOS expression. The inhibitory effect of p38mapk appears more complex and may be due to the ability of p38mapk to inhibit LPS-induced JNK activation. These results indicate that the MAPKs are important regulators of iNOS-NO· expression by IFN-gamma  + LPS.

nitric oxide; monocytes/macrophages; protein kinases; lipopolysaccharide; rodent


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