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Tumor necrosis factor increases the production of plasminogen activator
inhibitor in human endothelial cells in vitro and in rats in vivo
VW van Hinsbergh, T Kooistra, EA van den Berg, HM Princen, W Fiers and JJ Emeis
Gaubius Institute TNO, Leiden, The Netherlands.
The vascular endothelium plays an important role in fibrinolysis by
producing tissue-type plasminogen activator (t-PA) and plasminogen
activator inhibitor (PAI). The monokine tumor necrosis factor (human
recombinant TNF) increased the production of PAI by cultured human
endothelial cells from umbilical vein (twofold) and from foreskin
microvessles (four to eight fold). This was demonstrated by titration of
endothelial cell-conditioned medium with t-PA, by reverse fibrin
autography, and by immunoprecipitation of [35S]PAI-1 by anti-PAI-1 IgG. TNF
also induced a marked increase of PAI-1 messenger RNA (mRNA) in the cells.
The stimulation of PAI activity by TNF was seen at 4 U/mL and reached a
maximum at 500 U/mL. Human recombinant lymphotoxin and interleukin-1 (alpha
and beta) also stimulated the production of PAI activity, while
interleukin-6 was ineffective. Separate additions of TNF or interleukin-1
(IL-1) at optimal concentrations (500 U/mL and 5 U/mL, respectively)
resulted in a comparable stimulation of PAI production by endothelial
cells. The simultaneous addition of both mediators resulted in an additive
effect. The effect of TNF could not be prevented by the addition of
polymyxin B or by anti-IL-1 antibodies. Therefore, it is unlikely that TNF
acts through the induction of IL-1 secretion by endothelial cells. Two
hours after a bolus injection of 250,000 U/kg TNF into rats, a fivefold
increase in circulating PAI levels was found. In the next ten hours, the
levels returned to normal. Blood platelets do not significantly contribute
to the increase in circulating PAI, because the number of platelets did not
change after TNF injection and the amount of PAI in blood platelets is not
sufficient for several hours during an increase in PAI activity. The acute
phase reactants, fibrinogen and alpha 2-antiplasmin in rat plasma, were
altered little if any two to 24 hours after injection of 250,000 U/kg TNF.
In vitro, TNF did not change PAI production by human and rat hepatocytes in
primary monolayer culture. Therefore, it is most likely that vascular
endothelial cells contribute to the increased amount of circulating PAI
induced by TNF in vivo. This increase in PAI activity might decrease
fibrinolysis.
Volume 72,
Issue 5,
pp. 1467-1473,
11/01/1988
Copyright © 1988 by The American Society of Hematology
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