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Cytochrome P450IIIA activity and cytokine-mediated synthesis of nitric oxide.

Publication ,  Journal Article
Kuo, PC; Abe, KY; Dafoe, DC
Published in: Surgery
August 1995

BACKGROUND: Although nitric oxide synthase (NOS) is a cytochrome P450-like hemoprotein with additional sequence homology to cytochrome P450 reductase, the role of the cytochrome P450 system in cytokine-mediated NO synthesis is unknown. METHODS: To characterize the role of the P450 system in the synthesis of NO, NO production, NOS enzyme activity, and steady state NOS mRNA and protein expression were characterized in the setting of P450 isoform activity inhibition by using a model of isolated rat hepatocytes in primary culture. Cimetidine (0 to 10 mmol/L) was chosen as a specific inhibitor of P450IIIA activity. NO production was induced by interleukin-1 (50 ng/ml) and tumor necrosis factor (500 units/ml) and quantified by measurement of its metabolite, nitrite, in the culture medium. Steady state NOS mRNA and protein expression were determined by reverse-transcriptase polymerase chain reaction and immunoblot analysis, respectively. NOS enzyme activity was measured by the conversion of tritiated-L-arginine to tritiated-L-citrulline. RESULTS: Inhibition of P450IIIA activity was associated with a concentration-dependent decrease in cytokine-mediated NO production. Levels of NOS mRNA and protein were not altered. The NOS enzyme assay was notable for stable concentrations of intermediate, N-OH-L-arginine, and decreased production of the final end product, L-citrulline. Dixon plot kinetic analysis of cimetidine-mediated inhibition of NOS yielded an inhibition constant Ki = 1.76 mmol/L. CONCLUSIONS: These results indicate that cytochrome P450IIIA isoform may play a posttranslational role in cytokine-mediated NO synthesis in this model of isolated rat hepatocytes in primary culture.

Duke Scholars

Published In

Surgery

DOI

ISSN

0039-6060

Publication Date

August 1995

Volume

118

Issue

2

Start / End Page

310 / 317

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Surgery
  • Rats, Inbred Lew
  • Rats
  • RNA, Messenger
  • Polymerase Chain Reaction
  • Nitric Oxide Synthase
  • Nitric Oxide
  • Molecular Sequence Data
  • Molecular Probes
 

Citation

APA
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ICMJE
MLA
NLM
Kuo, P. C., Abe, K. Y., & Dafoe, D. C. (1995). Cytochrome P450IIIA activity and cytokine-mediated synthesis of nitric oxide. Surgery, 118(2), 310–317. https://doi.org/10.1016/s0039-6060(05)80339-3
Kuo, P. C., K. Y. Abe, and D. C. Dafoe. “Cytochrome P450IIIA activity and cytokine-mediated synthesis of nitric oxide.Surgery 118, no. 2 (August 1995): 310–17. https://doi.org/10.1016/s0039-6060(05)80339-3.
Kuo PC, Abe KY, Dafoe DC. Cytochrome P450IIIA activity and cytokine-mediated synthesis of nitric oxide. Surgery. 1995 Aug;118(2):310–7.
Kuo, P. C., et al. “Cytochrome P450IIIA activity and cytokine-mediated synthesis of nitric oxide.Surgery, vol. 118, no. 2, Aug. 1995, pp. 310–17. Pubmed, doi:10.1016/s0039-6060(05)80339-3.
Kuo PC, Abe KY, Dafoe DC. Cytochrome P450IIIA activity and cytokine-mediated synthesis of nitric oxide. Surgery. 1995 Aug;118(2):310–317.
Journal cover image

Published In

Surgery

DOI

ISSN

0039-6060

Publication Date

August 1995

Volume

118

Issue

2

Start / End Page

310 / 317

Location

United States

Related Subject Headings

  • Transcription, Genetic
  • Surgery
  • Rats, Inbred Lew
  • Rats
  • RNA, Messenger
  • Polymerase Chain Reaction
  • Nitric Oxide Synthase
  • Nitric Oxide
  • Molecular Sequence Data
  • Molecular Probes