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1
Institute of Parasitology, McGill University, Ste. Anne de Bellevue, Quebec, Canada
The pathogenesis of septicemia can be triggered by LPS, a potent
stimulus for PG synthesis. The enzyme cyclooxygenase (COX) is a
rate-limiting step in PG production. COX exists as two isoforms: COX-1,
which is constitutively expressed in most cell types, and COX-2, which
is inducible by LPS and cytokines in a variety of cells. In this study
we determined the role of the proinflammatory cytokines IL-1ß and
TNF-
released by LPS-stimulated U937 human macrophages in the
regulation of COX-2. Macrophages exposed to LPS showed a rapid and
sustained expression of COX-2 mRNA and protein for up to 48 h,
whereas PGE2 production was notably enhanced only after
12 h. LPS increased COX-2 gene transcription and
activation of the transcription factor NF-
B in a transient manner.
LPS-treated macrophages produced high levels of TNF- and moderate
amounts of IL-1ß protein. However, neutralizing Abs against these
cytokines had no effect on COX-2 mRNA and protein expression, nor did
they affect the stability of COX-2 mRNA. Interestingly, in the presence
of LPS or exogenous IL-1ß, COX-2 transcripts were stabilized, and
actinomycin D inhibited their degradation. Only when LPS or IL-1ß was
removed did COX-2 mRNA decay with a t1/2 of
5 h. In contrast, dexamethasone promoted a faster decay of the
LPS-induced COX-2 transcripts (t1/2 =
2.5 h). These results clearly demonstrate that LPS can regulate
COX-2 at both transcriptional and posttranscriptional levels
independently from endogenous IL-1ß and TNF- in human
macrophages.
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