Antioxidants increase lipopolysaccharide-stimulated TNFα release in murine macrophages: Role for altered TNFα mRNA stability

Avery B. Nathens, Richard Bitar, John C. Marshall, Ronald W G Watson, Alan P B Dackiw, Jie Fan, John Hiscott, Ori D. Rotstein

Research output: Contribution to journalArticle

Abstract

Through their effects on gene activation, antioxidants have been reported to modulate cellular expression of several proinflammatory cytokines and adhesion molecules, an effect mediated by preventing translocation of the transcription factor nuclear factor-κB (NF-κB) into the nucleus. In addition, modulation of the intracellular redox state may have profound effects on cell activation and subsequent gene expression distinct from effects on NF-κB; these effects may account for the divergent effects of antioxidants on cytokine gene expression in various reports. In the present studies, we evaluated the effect of the antioxidant, pyrrolidine dithiocarbamate (PDTC), on murine and human myeloid cell tumor necrosis factor alpha (TNFα) gene and protein expression. PDTC-enhanced LPS-induced TNFα secretion in cells derived from a murine macrophage cell line (J774.1), as well as in primary murine peritoneal macrophages by 4-fold. The effect was both stimulus and species dependent, as TNFα secretion was attenuated by PDTC in human THP-1 cells and in murine cells stimulated with zymosan. Northern analysis demonstrated that these effects were evident at the level of mRNA expression. Electrophoretic mobility shift assays confirmed the down-regulatory effect of PDTC on human myeloid NF-κB activation, whereas in murine cells no such inhibitory effect was evident. Evaluation of TNFα mRNA stability in murine cells demonstrated that the potentiating effect of PDTC on TNFα mRNA expression was due to an increase in mRNA half-life from 37 to 93 min. Together, these data suggest that the effect of antioxidants on gene expression are both stimulus and species dependent and illustrate a novel mechanism whereby redox manipulation might modulate TNFα expression in vivo.

Original languageEnglish (US)
Pages (from-to)361-367
Number of pages7
JournalShock
Volume16
Issue number5
StatePublished - Nov 2001
Externally publishedYes

Fingerprint

RNA Stability
Lipopolysaccharides
Tumor Necrosis Factor-alpha
Antioxidants
Macrophages
Gene Expression
Messenger RNA
Oxidation-Reduction
Cytokines
Zymosan
Peritoneal Macrophages
Electrophoretic Mobility Shift Assay
Myeloid Cells
Transcriptional Activation
Half-Life
Transcription Factors
pyrrolidine dithiocarbamic acid
Cell Line
Proteins

Keywords

  • Antioxidants
  • Dithiocarbamates
  • N-acetylcysteine
  • NF-κB

ASJC Scopus subject areas

  • Physiology
  • Critical Care and Intensive Care Medicine

Cite this

Nathens, A. B., Bitar, R., Marshall, J. C., Watson, R. W. G., Dackiw, A. P. B., Fan, J., ... Rotstein, O. D. (2001). Antioxidants increase lipopolysaccharide-stimulated TNFα release in murine macrophages: Role for altered TNFα mRNA stability. Shock, 16(5), 361-367.

Antioxidants increase lipopolysaccharide-stimulated TNFα release in murine macrophages : Role for altered TNFα mRNA stability. / Nathens, Avery B.; Bitar, Richard; Marshall, John C.; Watson, Ronald W G; Dackiw, Alan P B; Fan, Jie; Hiscott, John; Rotstein, Ori D.

In: Shock, Vol. 16, No. 5, 11.2001, p. 361-367.

Research output: Contribution to journalArticle

Nathens, AB, Bitar, R, Marshall, JC, Watson, RWG, Dackiw, APB, Fan, J, Hiscott, J & Rotstein, OD 2001, 'Antioxidants increase lipopolysaccharide-stimulated TNFα release in murine macrophages: Role for altered TNFα mRNA stability', Shock, vol. 16, no. 5, pp. 361-367.
Nathens AB, Bitar R, Marshall JC, Watson RWG, Dackiw APB, Fan J et al. Antioxidants increase lipopolysaccharide-stimulated TNFα release in murine macrophages: Role for altered TNFα mRNA stability. Shock. 2001 Nov;16(5):361-367.
Nathens, Avery B. ; Bitar, Richard ; Marshall, John C. ; Watson, Ronald W G ; Dackiw, Alan P B ; Fan, Jie ; Hiscott, John ; Rotstein, Ori D. / Antioxidants increase lipopolysaccharide-stimulated TNFα release in murine macrophages : Role for altered TNFα mRNA stability. In: Shock. 2001 ; Vol. 16, No. 5. pp. 361-367.
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