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Superoxide enhances interleukin 1beta-mediated transcription of the hepatocyte-inducible nitric oxide synthase gene.

Publication ,  Journal Article
Kuo, PC; Abe, K; Schroeder, RA
Published in: Gastroenterology
March 2000

BACKGROUND & AIMS: Exposure to oxidative stress, as in states of shock, ischemia-reperfusion injury, or sepsis, commonly initiates a complex cellular cascade of interlocking redox modulatory systems that detoxify electrophiles. In interleukin 1beta (IL-1beta)-treated rat hepatocytes, we have previously demonstrated that inducible nitric oxide synthase (iNOS) protein expression, steady-state iNOS messenger RNA (mRNA) levels, and NO synthesis are increased by oxidative stress induced by superoxide. The effect of hepatocellular redox state upon iNOS gene transcription has not been previously studied. METHODS: Using rat hepatocytes in primary culture, iNOS gene transcription was induced by IL-1beta. Oxidative stress was mediated by 1,2,3-benzenetriol (BZT), an autocatalytic source of superoxide. Nuclear run-on assays and transient transfection assays using the rat hepatocyte iNOS full-length promoter and deletion constructs were designed to isolate a cis-acting regulatory element. Specificity was confirmed by site-directed mutagenesis. Gel shift analysis determined the presence of a corresponding trans-acting regulatory factor. RESULTS: In IL-1beta-treated cells, BZT increased iNOS gene transcription without altering mRNA half-life. An antioxidant-responsive element (ARE) was found in the iNOS promoter at base pair -1347, which conferred redox sensitivity. Gel shift analysis identified a corresponding nuclear protein capable of binding to ARE in IL-1beta- and BZT-treated rat hepatocytes. CONCLUSIONS: An ARE in the rat hepatocyte iNOS promoter confers redox sensitivity and augments IL-1beta-mediated iNOS gene and protein expression in the setting of superoxide treatment.

Duke Scholars

Published In

Gastroenterology

DOI

ISSN

0016-5085

Publication Date

March 2000

Volume

118

Issue

3

Start / End Page

608 / 618

Location

United States

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Superoxides
  • Response Elements
  • Rats, Inbred Lew
  • Rats
  • Pyrogallol
  • Promoter Regions, Genetic
  • Oxidative Stress
  • Nitric Oxide Synthase Type II
 

Citation

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Kuo, P. C., Abe, K., & Schroeder, R. A. (2000). Superoxide enhances interleukin 1beta-mediated transcription of the hepatocyte-inducible nitric oxide synthase gene. Gastroenterology, 118(3), 608–618. https://doi.org/10.1016/s0016-5085(00)70268-x
Kuo, P. C., K. Abe, and R. A. Schroeder. “Superoxide enhances interleukin 1beta-mediated transcription of the hepatocyte-inducible nitric oxide synthase gene.Gastroenterology 118, no. 3 (March 2000): 608–18. https://doi.org/10.1016/s0016-5085(00)70268-x.
Kuo, P. C., et al. “Superoxide enhances interleukin 1beta-mediated transcription of the hepatocyte-inducible nitric oxide synthase gene.Gastroenterology, vol. 118, no. 3, Mar. 2000, pp. 608–18. Pubmed, doi:10.1016/s0016-5085(00)70268-x.
Journal cover image

Published In

Gastroenterology

DOI

ISSN

0016-5085

Publication Date

March 2000

Volume

118

Issue

3

Start / End Page

608 / 618

Location

United States

Related Subject Headings

  • Transfection
  • Transcription, Genetic
  • Superoxides
  • Response Elements
  • Rats, Inbred Lew
  • Rats
  • Pyrogallol
  • Promoter Regions, Genetic
  • Oxidative Stress
  • Nitric Oxide Synthase Type II