Transcriptional up-regulation of intracellular adhesion molecule-1 in human endothelial cells by the antioxidant pyrrolidine dithiocarbamate involves the activation of activating protein-1

J Immunol. 1996 Oct 15;157(8):3587-97.

Abstract

The redox status of the cell plays an essential role in regulating signal transduction, transcription factor activity, and expression of cell surface molecules. In this study, we show that pyrrolidine dithiocarbamate (PDTC), a potent antioxidant agent, upregulated the cell surface expression of intercellular adhesion molecule-1 (ICAM-1) in human endothelial cells (EC). Further analysis of PDTC-mediated ICAM-1 up-regulation revealed that PDTC increased ICAM-1 mRNA levels and augmented its gene promoter activity. Transfection experiments in EC with reporter constructs harboring nested deletion fragments of the ICAM-1 promoter indicated the presence of a functional PDTC-responsive region located between positions -136 to -353 of the promoter. Gel retardation assays together with supershift analysis revealed that PDTC induced the binding of c-fos and c-jun to a consensus activating protein-1 (AP-1) binding site located at position -284. PDTC alone or in combination with TNF-alpha enhanced AP-1-dependent transactivation in HUVEC, as determined by DNA binding assays. The functional implication of AP-1 in the transcription of the ICAM-1 gene was further demonstrated by cotransfection experiments in which a c-jun expression vector induced the promoter activity of the PDTC-responsive element of the ICAM-1 promoter. Taken together, these results indicate that the antioxidant PDTC induces transcriptional activation of ICAM-1 and that this induction is mediated at least in part by the transcription factor AP-1. This mechanism might be operative in pathologic conditions in which a redox imbalance plays a key role, such as ischemia/reperfusion injury or arteriosclerosis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antioxidants / pharmacology*
  • Base Sequence
  • Cells, Cultured
  • DNA / genetics
  • DNA / metabolism
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects*
  • Endothelium, Vascular / metabolism*
  • Humans
  • Inflammation Mediators / pharmacology
  • Intercellular Adhesion Molecule-1 / genetics*
  • Oligonucleotide Probes / genetics
  • Oxidation-Reduction
  • Promoter Regions, Genetic / drug effects
  • Pyrrolidines / pharmacology*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Thiocarbamates / pharmacology*
  • Transcription Factor AP-1 / metabolism*
  • Transcriptional Activation / drug effects
  • Transfection
  • Tumor Necrosis Factor-alpha / pharmacology
  • Up-Regulation / drug effects

Substances

  • Antioxidants
  • Inflammation Mediators
  • Oligonucleotide Probes
  • Pyrrolidines
  • RNA, Messenger
  • Thiocarbamates
  • Transcription Factor AP-1
  • Tumor Necrosis Factor-alpha
  • Intercellular Adhesion Molecule-1
  • pyrrolidine dithiocarbamic acid
  • DNA