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Prepublished online as a Blood First Edition Paper on May 1, 2003; DOI 10.1182/blood-2002-11-3607.
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Blood, 1 September 2003, Vol. 102, No. 5, pp. 1693-1700
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Synergistic effect of sphingosine 1-phosphate on thrombin-induced tissue factor expression in endothelial cells
Hiroyuki Takeya,
Esteban C. Gabazza,
Shinya Aoki,
Hikaru Ueno, and
Koji Suzuki
From the Department of Molecular Pathobiology, Mie University School of
Medicine, Tsu, Japan; and the Department of Biochemistry and Molecular
Pathophysiology, University of Occupational and Environmental Health, School
of Medicine, Kitakyushu, Japan.
Sphingosine 1-phosphate (S1P), a bioactive lipid, is produced and stored in
platelets and is released from activated platelets during blood coagulation
activation. Thrombin, which is also generated during blood coagulation, has
been shown to induce tissue factor (TF), the initiator of blood coagulation,
in endothelial cells (ECs); however, the effect of S1P on this process is not
evaluated. Here we demonstrated that S1P strongly potentiated thrombin-induced
TF expression in ECs and that S1P itself did not induce TF expression. Among
signaling lipids, platelet-activating factor slightly enhanced
thrombin-induced TF expression; other lipids, including lysophosphatidic acid,
lysophosphatidylcholine, sphingosine, and C2-ceramide exert no
effect on TF expression. S1P enhanced TF expression at the transcriptional
level, possibly via promoting the activation of transcription factors nuclear
factor B (NF- B) and Egr-1. Thrombin weakly and S1P
strongly activated extracellular signalregulated kinase 1/2 (ERK1/2)
mitogen-activated protein (MAP) kinase and, in the presence of both
stimulants, enhanced and sustained activation of this kinase was observed. The
ERK1/2-specific inhibitor PD98059 significantly inhibited enhanced TF
expression induced by both stimulants but only weakly inhibited
thrombin-induced TF expression, thus indicating the requirement of the ERK1/2
pathway in synergistic induction of TF expression. In addition, we found that
thrombin and S1P rapidly up-regulated the expression of S1P receptors,
endothelial differentiation gene-1 (EDG-1) and EDG-3, thereby suggesting that
the effect of S1P on TF expression and other EC functions may be enhanced by
thrombin and S1P itself. The present data reveal the synergistic effect of S1P
on thrombin-induced TF expression in ECs, which may promote further thrombin
and S1P generation, thus propagating a positive feedback reaction.

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