Up-Regulation of Protein Kinase C-ϵ Promotes the Expression of Cytokine-inducible Nitric Oxide Synthase in RAW 264.7 Cells

Díaz-Guerra, María José; Bodelón, Oscar G.; Velasco, Marta; Whelan, Richard D. H.; Parker, Peter J.; Boscá, Lisardo

doi:10.1074/jbc.271.50.32028

Cited by 68 publications

(42 citation statements)

References 46 publications

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“…RAW 264.7 cells were established from peritoneal macrophages, but the cells have been transformed and immortalized by infection of Abelson leukemia virus (16). Although peritoneal macrophages and RAW 264.7 cells share many biological features, these cells are reported to elicit different responses in NO synthesis when stimulated with phorbol ester (37). We also observed in this study that AM increased cAMP production in peritoneal macrophages but not in RAW 264.7 cells.…”

Section: Discussionsupporting

confidence: 69%

Production of Adrenomedullin in Macrophage Cell Line and Peritoneal Macrophage

Kubo¹,

Minamino²,

Isumi³

et al. 1998

Journal of Biological Chemistry

163

111

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We demonstrate that adrenomedullin (AM) is produced and secreted from cultured murine monocyte/ macrophage cell line (RAW 264.7) as well as mouse peritoneal macrophage. Immunoreactive (IR) AM secreted from RAW 264.7 cells was chromatographically identified to be native AM. To elucidate the regulation mechanism of AM production in macrophage, we examined the effects of various substances inducing differentiation or activation of monocyte/macrophage. Phorbol ester (TPA), retinoic acid (RA), lipopolysaccharide (LPS), and interferon-␥ (IFN-␥) increased AM production 1.5-7-fold in RAW 264.7 cells in a dose-as well as time-dependent manner. By LPS stimulation, the AM mRNA level in RAW 264.7 cells was augmented up to 7-fold after 14 h incubation. RA exerted a synergistic effect when administered with TPA, LPS, or IFN-␥, whereas IFN-␥ completely suppressed AM production in RAW 264.7 cells stimulated with LPS. Dexamethasone, hydrocortisone, estradiol, and transforming growth factor-␤ dosedependently suppressed AM production in RAW 264.7 cells. AM production was also investigated in mouse peritoneal macrophage. Primary mouse macrophage secreted IR-AM at a rate similar to that of RAW 264.7 cells, and its production was enhanced 9-fold by LPS stimulation. AM was found to increase basal secretion of tumor necrosis factor ␣ (TNF-␣) from RAW 264.7 cells, whereas AM suppressed the secretion of TNF-␣ and interleukin-6 from that stimulated with LPS. Thus, macrophage should be recognized as one of the major sources of AM circulating in the blood. Especially in cases of sepsis and inflammation, AM production in macrophage is augmented, and the secreted AM is deduced to function as a modulator of cytokine production. Adrenomedullin (AM)1 is a potent vasorelaxant peptide originally isolated from extracts of human pheochromocytoma by monitoring the elevating activity of platelet cAMP (1). AM shows slight homology with calcitonin gene-related peptide (CGRP) and has a potent and long-lasting depressor effect when injected intravenously into anesthetized rats (1, 2). We have shown that cultured endothelial cells (ECs) and vascular smooth muscle cells (VSMCs) produce and secrete AM into culture medium (3, 4). The production and secretion of AM in VSMC and EC were augmented by interleukin-1 (IL-1), tumor necrosis factor ␣ (TNF-␣), and lipopolysaccharide (LPS) (5, 6), which are known to be major factors inducing septic shock (7-9). In the in vivo study, intravenous administration of LPS into rats actually elevated plasma AM concentration 20-fold and augmented AM gene expression in blood vessels, lung, and intestine (10). Plasma AM levels were also remarkably increased in patients with septic shock compared with those in healthy volunteers (11,12). These data suggest the possibility that AM contributes to induction of refractory hypotension in septic shock. On the other hand, macrophages are activated by exposure to stimuli of foreign bodies such as LPS and then start to produce and secrete various cytokines, such as IL-1 and TNF-␣. These da...

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Section: Discussionsupporting

confidence: 69%

Production of Adrenomedullin in Macrophage Cell Line and Peritoneal Macrophage

Kubo¹,

Minamino²,

Isumi³

et al. 1998

Journal of Biological Chemistry

163

111

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“…The ability to elevate p65 transactivation was most apparent with the novel isoforms, particularly PKC⑀, suggesting that these kinases may be responsible for regulating transactivation. Similarly, PKC⑀ has previously been shown to activate NF-B-dependent transcription in RAW 264.7 cells providing support for a role of this kinase (53).…”

Section: Discussionmentioning

confidence: 99%

Inhibitors of Protein Kinase C (PKC) Prevent Activated Transcription

Catley

Cambridge

Nasuhara

et al. 2004

Journal of Biological Chemistry

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“…These reports provide information on signaling pathways for NOS-2 expression at 4 -6 or 20 -24 h but did not provide kinetic data. For example, Salh et al (25) report the inhibition of NOS-2 expression at 6 h by LY294002, whereas Diaz-Guerra et al (26) and Chen et al (27) report the involvement of PKC in NOS-2 expression at 24 h. Diaz-Guerra et al (28) report that NO production stimulated by LPSϩIFN-␥ for 24 h was augmented by LY294002. These results are consistent with our findings that PI3K mediates NOS-2 expression at 4 -6 h and negatively regulates its expression at 24 h. Taken together, these results support the notion that NOS-2 expression stimulated by LPSϩIFN-␥ comprises two distinct phases that are signaled by different pathways: 1) an early phase of expression, which is signaled through PI3K, p38 MAPK, and Jak 2/3; and 2) a late phase of expression, which is signaled via PKC and mammalian target of rapamycin.…”

Section: Nos-2 and Cox-2 Expression At 4 H Was Mediated By A Similarmentioning

confidence: 99%

Salicylate Suppresses Macrophage Nitric-oxide Synthase-2 and Cyclo-oxygenase-2 Expression by Inhibiting CCAAT/Enhancer-binding Protein-β Binding via a Common Signaling Pathway

Cieslik

Zhu

2002

Journal of Biological Chemistry

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We determined whether salicylate at pharmacological concentrations inhibits nitric-oxide synthase-2 (NOS-2) and cyclo-oxygenase-2 (COX-2) expressions in RAW 264.7 stimulated with lipopolysaccharide (LPS) and interferon-γ (IFN-γ). Cells were treated with sodium salicylate (10−7-10−4m) or vehicle for 30 min followed by LPS+IFN-γ for up to 24 h. Salicylate suppressed NOS-2 and COX-2 protein levels and promoter activities stimulated by LPS+IFN-γ for 4 h in a concentration-dependent manner but had no effect on NOS-2 expression stimulated by the combined agonists for 24 h. Results from promoter analysis indicate that the binding of CCAAT/enhancer-binding protein β (C/EBPβ) to its cognate site at −150/−142 on the NOS-2 promoter region was essential for NOS-2 expression at 4 h but not at 24 h. Salicylate reduced C/EBPβ binding at 4 h and did not alter its binding at 24 h. NOS-2 and COX-2 protein levels and C/EBPβ binding stimulated by LPS+IFN-γ for 4 h were inhibited by a similar battery of signaling inhibitors, suggesting a common pathway for NOS-2 and COX-2 expression. Kinetic analysis indicates that NOS-2, similar to COX-2 expression, at 4 h was largely due to the action of LPS, which induced C/EBPβ binding, whereas its expression at a longer time point was contributed by IFN-γ. Our findings implicate two distinct pathways for NOS-2 expression induced by LPS+IFN-γ. Salicylate at pharmacological concentrations is capable of suppressing the early phase of NOS-2 and COX-2 expression by blocking C/EBPβ binding.

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Up-Regulation of Protein Kinase C-ϵ Promotes the Expression of Cytokine-inducible Nitric Oxide Synthase in RAW 264.7 Cells

Cited by 68 publications

References 46 publications

Production of Adrenomedullin in Macrophage Cell Line and Peritoneal Macrophage

Production of Adrenomedullin in Macrophage Cell Line and Peritoneal Macrophage

Inhibitors of Protein Kinase C (PKC) Prevent Activated Transcription

Salicylate Suppresses Macrophage Nitric-oxide Synthase-2 and Cyclo-oxygenase-2 Expression by Inhibiting CCAAT/Enhancer-binding Protein-β Binding via a Common Signaling Pathway

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