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Vol. 273, Issue 3, 1733-1740, January 16, 1998
From the Department of Health Chemistry, School of Pharmaceutical
Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa-ku,
Tokyo 142, Japan
Activation of rat fibroblastic 3Y1 cells with
interleukin-1
Cytosolic Phospholipase A2 Is Required for
Cytokine-induced Expression of Type IIA Secretory Phospholipase
A2 That Mediates Optimal
Cyclooxygenase-2-dependent Delayed Prostaglandin
E2 Generation in Rat 3Y1 Fibroblasts
(IL-1) and tumor necrosis factor 
(TNF)
induced delayed prostaglandin (PG) E2 generation over
6-48 h, which occurred in parallel with de novo induction
of type IIA secretory phospholipase A2 (sPLA2) and cyclooxygenase (COX)-2, without accompanied by changes in the
constitutive expression of type IV cytosolic PLA2
(cPLA2) and COX-1. Types V and IIC sPLA2s were
barely detectable in these cells. Studies using an anti-type IIA
sPLA2 antibody, sPLA2 inhibitors, and a type
IIA sPLA2-specific antisense oligonucleotide revealed that
IL-1/TNF-induced delayed PGE2 generation by these
cells was largely dependent on inducible type IIA sPLA2,
which was functionally linked to inducible COX-2. Delayed
PGE2 generation was also suppressed markedly by the
cPLA2 inhibitor arachidonoyl trifluoromethyl ketone (AACOCF3), which attenuated induction of type IIA
sPLA2, but not COX-2, expression. AACOCF3
inhibited the initial phase of cytokine-stimulated arachidonic acid
release, and supplementing AACOCF3-treated cells with
exogenous arachidonic acid partially restored type IIA
sPLA2 expression. These results suggest that certain
metabolites produced by the cPLA2-dependent
pathway are crucial for the subsequent induction of type IIA
sPLA2 expression and attendant delayed PGE2 generation. Some lipoxygenase-derived products might be involved in
this event, since IL-1/TNF-induced type IIA sPLA2
induction and PGE2 generation were reduced markedly by
lipoxygenase, but not COX, inhibitors. In contrast, Ca2+
ionophore-stimulated immediate PGE2 generation was
regulated predominantly by the constitutive enzymes cPLA2
and COX-1, even when type IIA sPLA2 and COX-2 were
maximally induced after IL-1/TNF treatment, revealing functional
segregation of the constitutive and inducible PG biosynthetic
enzymes.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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