Pyrrolidine Dithiocarbamate Inhibits Induction of Nitric Oxide Synthase Activity in Rat Alveolar Macrophages

Sherman, Michael P.; Aeberhard, Ernesto E.; Wong, Vivian; Griscavage, Jeanette M.; Ignarro, Louis J.

doi:10.1006/bbrc.1993.1359

Cited by 236 publications

(142 citation statements)

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“…The lavage effluent was centrifuged at 250 x g at 8°C for 10 min. The cell pellets were resuspended in Hepes buffer with 0.3% bovine serum albumin, the cells were counted in a hemacytometer, and viability was measured by trypan blue exclusion (20,21). Cytopreps were stained with WrightGiemsa and cell differential counts were then performed.…”

Section: Methodsmentioning

confidence: 99%

“…Cytopreps were stained with WrightGiemsa and cell differential counts were then performed. (20). NR8383 macrophages were utilized as our positive control as these cells can be stimulated by cytokines in vitro to produce NO (20).…”

Section: Methodsmentioning

confidence: 99%

“…(20). NR8383 macrophages were utilized as our positive control as these cells can be stimulated by cytokines in vitro to produce NO (20). NO production was indirectly assessed by estimation of nitrite and nitrate concentration in the culture supernatant.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Nitric oxide is an important mediator for tumoricidal activity in vivo.

Farias‐Eisner

Sherman

Aeberhard

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Nitric oxide is an important mediator for tumoricidal activity in vivo.

Farias‐Eisner

Sherman

Aeberhard

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

215

122

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“…Amongst the 24 consensus sequences that have been identified in the promoter region of the murine iNOS gene are putative binding sites for transcription factors including NF-KB and NFIL-6 whose activity can be stimulated by protein kinase(s) A, protein kinase C and protein tyrosine kinase Ghosh & Baltimore, 1990;Metz & Ziff, 1991;Link et al, 1992). Although the role of NFIL-6 still remains to be clarified, activation of NF-KB seems to be a necessary event for the expression of iNOS in lipopolysaccharide-stimulated macrophages and in IL-lfl-activated vascular smooth muscle cells (Xie et al, 1993;Sherman et al, 1993;MUlsch et al, 1993;Schini-Kerth et al, 1994b). Therefore, the stimulating effect of cyclic AMP on iNOS expression in vascular smooth muscle cells could occur at the level of NF-KB activation.…”

Section: Expression Of Inos Proteinmentioning

confidence: 99%

Effect of cyclic GMP‐dependent vasodilators on the expression of inducible nitric oxide synthase in vascular smooth muscle cells: role of cyclic AMP

Boese

Busse

Mülsch

et al. 1996

British J Pharmacology

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1 In the present study we examined whether interleukin-lf (IL-Ifl) increases the activity of adenylyl cyclase in vascular smooth muscle cells and determined its role in the cytokine-induced expression of the inducible nitric oxide synthase (iNOS) and activation of nuclear transcription factor-KB (NF-KB). In addition the interaction between cyclic AMP-and cyclic GMP-elevating agonists on the IL-iIf-stimulated expression of iNOS was examined.2 Exposure of vascular smooth muscle cells to IL-,IB stimulated the formation of cyclic AMP but not of cyclic GMP. The intracellular level of cyclic AMP reached a maximum within 1 h and then gradually declined over the next 5 h. This IL-l, (60 u ml-')-stimulated formation of cyclic AMP was modest (about 3 fold at 60 u ml-' for 1 h) compared to that evoked by isoprenaline (about 9 fold at 3 x 10-6 M for 2 min). 3 The IL-iI (60 u ml1 for 24 h)-stimulated accumulation of nitrite, which was taken as an index of NO production, was concentration-dependently increased by preferential inhibitors of cyclic AMP-dependent phosphodiesterases (rolipram and trequinsin). This effect was reproduced by a specific activator of the cyclic AMP-dependent protein kinase(s) A, Sp-8-CPT-cAMPS (10-4 M) but was prevented by a specific inhibitor of cyclic AMP-dependent protein kinase(s) A, Rp-8-CPT-cAMPS (10-4 M). These compounds alone [rolipram (10-6 M), trequinsin (3 x 10-6 M) and Sp-8-CPT-cAMPS (l0-4 M)] slightly but significantly increased the release of nitric oxide while Rp-8-CPT-cAMPS elicited no such effect. 4 Inducible NOS protein was expressed in IL-/IB (30 u ml-', 24 h)-stimulated smooth muscle cells as assessed by Western blot analysis. The level of iNOS protein was markedly increased in smooth muscle cells which had been exposed to IL-,IB in combination with either rolipram (3 x 10-6 M) or Sp-8-CPTcAMPS (l0-4 M) but was reduced in those exposed to IL-,IB and Rp-8-CPT-cAMPS (l0-4 M). A weak expression of iNOS protein was found in smooth muscle cells which had been exposed to either Sp-8-CPT-cAMPS or rolipram alone for 24 h while Rp-8-CPT-cAMPS elicited no such effect. 5 Exposure of smooth muscle cells to IL-lI (30 u ml-') for 30 min increased the level of NF-KB-DNA complexes in nuclear extracts as detected by electrophoretic mobility shift assay. Similar levels of NF-KB-DNA complexes were found in cells which had been exposed to IL-,IB in combination with either Sp

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“…In the present study, we provide evidence for the involvement of several factors binding within the rat Nos2 core promoter region. Previous studies in the mouse have implicated NF-κB (Lowenstein, et al, 1993, Sherman, et al, 1993 and IRF-1 (Kamijo, et al, 1994, Martin, et al, 1994 in the transcriptional response of the Nos2 gene to LPS+IFNγ. In the rat, NF-κB, C/ EBPβ and ATF-2, and CREB (Bhat, et al, 2002, Gavrilyuk, et al, 2001) have been implicated in transcriptional activation of Nos2 by LPS+IFNγ in glial cells.…”

Section: Discussionmentioning

confidence: 99%

The Oct DNA motif participates in the alcohol inhibition of the inducible nitric oxide synthase gene promoter in rat C6 glioma cells

2007

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Induction of nitric-oxide synthase-2 (iNOS) by cytokines and bacterial products is associated with protein binding at the proximal promoter and in an upstream enhancer region of the Nos2 gene. To clarify how ethanol suppresses rat iNOS activity, we constructed several deletion mutants of the Nos2 promoter fused to the luciferase gene and transfected the constructs into C6 glial cells. Acute ethanol exposure of stably transfected cells for 24 h inhibits induced activity of Nos2 promoter constructs containing deletions in the 5′ flanking region, including a 94 bp promoter that lacks any known NF-κB site but which carries a C/EBPβ and overlapping γ-IRE, GAS and Oct motifs. Ethanol failed to inhibit the endogenous activity of a smaller, 78 bp promoter that lacks the C/EBPβ and overlapping, γ-IRE and GAS motifs and showed no inducible activity. As another approach, in vivo DNA footprinting was used and identified protein protections at five regions of the proximal Nos2 promoter in induced cells. Exposure to acute ethanol diminished protein occupation in the five promoter regions including the γ-IRE/NF-κB and the overlapping γIRE/GAS/Oct sites. Site-directed mutagenesis in the octamer domain of the γIRE/GAS/Oct motifs was studied in a 1002 bp promoter to examine its role in ethanol inhibition of cytokine and lipopolysaccharide induced activity. The data indicate ethanol failed to inhibit promoter activity when the Oct motif is missing. Electrophoretic mobility shift assays performed using a 22-mer probe containing the overlapping γ-IRE/GAS/Oct sites showed three complexes with one of the complexes being competed by an octamer-1 antibody. These observations demonstrate the role of protein-DNA binding at the core promoter, and the likely involvement of the octamer motif, in ethanol modulation of cytokine and lipopolysaccharide induced iNOS expression.

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Pyrrolidine Dithiocarbamate Inhibits Induction of Nitric Oxide Synthase Activity in Rat Alveolar Macrophages

Cited by 236 publications

References 0 publications

Nitric oxide is an important mediator for tumoricidal activity in vivo.

Nitric oxide is an important mediator for tumoricidal activity in vivo.

Effect of cyclic GMP‐dependent vasodilators on the expression of inducible nitric oxide synthase in vascular smooth muscle cells: role of cyclic AMP

The Oct DNA motif participates in the alcohol inhibition of the inducible nitric oxide synthase gene promoter in rat C6 glioma cells

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