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Posttranscriptional regulation of macrophage tissue factor expression by
antioxidants
GF Brisseau, AP Dackiw, PY Cheung, N Christie and OD Rotstein
Department of Surgery, University of Toronto, Ontario, Canada.
Tissue factor (TF) expression by cells of monocyte/macrophage lineage
represents an important mechanism underlying the initiation of fibrin
deposition at sites of extravascular inflammation. Recent evidence suggests
a role for oxidant stress in the signalling pathway of various cell types
by virtue of its ability to induce DNA binding of various transcription
factors, including nuclear factor kappa B and AP-1. The effect of
antioxidant treatment on lipopolysaccharide (LPS)-induced TF expression was
examined in murine peritoneal macrophages and human monocytes. Both
pyrrolidine dithiocarbamate, an oxidant scavenger, and N-acetyl-cysteine, a
precursor of the endogenous antioxidant glutathione, inhibited stimulation
of macrophage procoagulant activity by LPS. Northern blot analysis showed
that neither of these agents reduced LPS-stimulated TF mRNA accumulation,
thereby suggesting a posttranscriptional mechanism for the effect.
Immunofluorescence studies of human monocytes using polyclonal anti-TF
antibody showed that N-acetyl-cysteine treatment prevented the
characteristic plasmalemmal localization of TF antigen that occurs in
response to LPS. Western blot analysis showed that N-acetyl-cysteine
reduced the accumulation of the 47-kD mature glycoprotein in LPS-treated
cells, a finding consistent with the results of the immunofluorescence
studies. Furthermore, these conditions did not result in an accumulation of
the less mature forms of TF. When considered together, these data suggest
that antioxidants exert their effects by impairing translation and/or by
causing degradation of newly translated protein. The effect of antioxidants
on tumor necrosis factor appeared to be species specific, with no effect on
LPS-induced tumor necrosis factor in murine cells, but with inhibition in
human monocytes. The posttranscriptional effect of antioxidants on TF
expression data suggests a novel mechanism whereby these agents might
modulate monocyte/macrophage activation.
Volume 85,
Issue 4,
pp. 1025-1035,
02/15/1995
Copyright © 1995 by The American Society of Hematology
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