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*
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan;
Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan; and
Discovery Research Laboratory I, Minase Research Institute, Ono Pharmaceutical, Osaka, Japan
The expression and regulation of the PGE receptors, EP2
and EP4, both of which are coupled to the stimulation of
adenylate cyclase, were examined in peritoneal resident macrophages
from C3H/HeN mice. mRNA expression of EP4 but not
EP2 was found in nonstimulated cells, but the latter was
induced by medium change alone, and this induction was augmented by
LPS. mRNA expression of EP4 was down-regulated by LPS but
not by medium change. PGE2 increased the cAMP content of
both LPS-treated and nontreated cells. ONO-604, an EP4
agonist, also increased cAMP content in nonstimulated cells and in
cells treated with LPS for 3 h, but not for 6 h. Butaprost,
an EP2 agonist, was effective only in the cells treated
with LPS for 6 h. The inhibitory effects of ONO-604 on TNF-
and
IL-12 production were equipotent with PGE2 at any time
point, but the inhibitory effects of butaprost were only seen from
14 h after stimulation. PGE2 or dibutyryl cAMP alone,
but not butaprost, reduced EP4 expression, and indomethacin
reversed the LPS-induced down-regulation of EP4, indicating
that the down-regulation of EP4 is mediated by LPS-induced
PG synthesis and EP4 activation. Indeed, when we used
C3H/HeJ (LPS-hyporesponsive) macrophages, such reduction in
EP4 expression was found in the cells treated with
PGE2 alone, but not in LPS-treated cells. In contrast,
up-regulation of EP2 expression was again observed in
LPS-treated C3H/HeJ macrophages. These results suggest that
EP4 is involved mainly in the inhibition of cytokine
release, and that the gene expression of EP2 and
EP4 is differentially regulated during macrophage
activation.
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