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J Biol Chem, Vol. 274, Issue 18, 12263-12268, April 30, 1999
From the Department of Chemistry and Biochemistry, School of
Medicine and Revelle College, University of California at San Diego,
La Jolla, California 92093-0601
Group V secretory phospholipase
A2 (sPLA2) rather than Group IIA
sPLA2 is involved in short term, immediate arachidonic acid mobilization and prostaglandin E2 (PGE2)
production in the macrophage-like cell line P388D1. When a
new clone of these cells, P388D1/MAB, selected on the basis
of high responsivity to lipopolysaccharide plus platelet-activating
factor, was studied, delayed PGE2 production (6-24 h) in
response to lipopolysaccharide alone occurred in parallel with the
induction of Group V sPLA2 and cyclooxygenase-2 (COX-2). No
changes in the level of cytosolic phospholipase A2
(cPLA2) or COX-1 were observed, and Group IIA
sPLA2 was not detectable. Use of a potent and selective
sPLA2 inhibitor, 3-(3-acetamide 1-benzyl-2-ethylindolyl-5-oxy)propanesulfonic acid (LY311727), and
an antisense oligonucleotide specific for Group V sPLA2
revealed that delayed PGE2 was largely dependent on the
induction of Group V sPLA2. Also, COX-2, not COX-1, was
found to mediate delayed PGE2 production because the
response was completely blocked by the specific COX-2 inhibitor NS-398.
Delayed PGE2 production and Group V sPLA2
expression were also found to be blunted by the inhibitor
methylarachidonyl fluorophosphonate. Because inhibition of
Ca2+-independent PLA2 by an antisense technique
did not have any effect on the arachidonic acid release, the data using
methylarachidonyl fluorophosphonate suggest a key role for the
cPLA2 in the response as well. Collectively, the results
suggest a model whereby cPLA2 activation regulates Group V
sPLA2 expression, which in turn is responsible for delayed
PGE2 production via COX-2.
Regulation of Delayed Prostaglandin Production in Activated
P388D1 Macrophages by Group IV Cytosolic and Group V
Secretory Phospholipase A2s
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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