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J. Biol. Chem., Vol. 276, Issue 8, 5467-5475, February 23, 2001
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From the Department of Microbiology and Immunology and
§ Internal Medicine, University of Texas Medical Branch,
Galveston, Texas 77555-1070
Prostaglandins (PG), which are responsible for a
large array of biological functions in eukaryotic cells, are produced
from arachidonic acid by phospholipases and cyclooxygenase enzymes COX-1 and COX-2. We demonstrated that PG levels in cells were partly
controlled by a regulatory protein, phospholipase A2
(PLA2)-activating protein (PLAA). Treatment of murine
macrophages with lipopolysaccharide, interleukin-1
Prostaglandin Levels in Stimulated Macrophages Are Controlled by
Phospholipase A2-activating Protein and by Activation of
Phospholipase C and D*
,
, and tumor
necrosis factor-
increased PLAA levels at early time points (2-30
min), which correlated with an up-regulation in cytosolic
PLA2 and PGE2 levels. Both COX-2 and secretory
PLA2 were also increased in lipopolysaccharide-stimulated
macrophages, however, at later time points of 4-24 h. The role of PLAA
in eicosanoid formation in macrophages was confirmed by the use of an
antisense plaa oligonucleotide. Within amino acid residues
503-538, PLAA exhibited homology with melittin, and increased
PGE2 production was noted in macrophages stimulated with
melittin. In addition to PLA2, we demonstrated that
activation of phospholipase C and D significantly controlled
PGE2 production. Finally, increased antigen levels of PLAA,
COX-2, and phospholipases were demonstrated in biopsy specimens from
patients with varying amounts of intestinal mucosal inflammation, which
corresponded to increased levels of phospholipase activity. These
results could provide a basis for the development of new therapeutic
tools to control inflammation.
*
This work was supported in part by a grant from the Crohn's
and Colitis Foundation of America and American Heart Association (to
A. K. C.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Supported by The James W. McLaughlin Predoctoral Fellowship Fund.
¶
To whom correspondence should be addressed: Dept. of
Microbiology and Immunology, 301 University Blvd., University of Texas Medical Branch, Galveston, TX 77555-1070. Tel.: 409-747-0578; Fax:
409-747-6869; E-mail: achopra@utmb.edu.
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