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Department of Pharmacology, New York Medical College, Valhalla, New York 10595
The medullary thick ascending limb (MTAL)
metabolizes arachidonic acid (AA) via cytochrome
P-450 (CyP450)- and cyclooxygenase (COX)-dependent pathways. In the present study, we demonstrated that
the COX-2-selective inhibitor, NS-398, prevented tumor necrosis factor-
(TNF)- and phorbol myristate acetate (PMA)-mediated
increases in PGE2 production by
cultured MTAL cells. Accumulation of COX-2, but not COX-1, mRNA
increased when cells were challenged with TNF (1 nM) or PMA (1 µM).
Pretreatment of cells for 30 min with actinomycin D (AcD, 1 µM) had
little effect on COX-2 mRNA accumulation in unstimulated cells or in
cells challenged with either TNF or PMA. Moreover, a
posttranscriptional mechanism(s) appears to contribute significantly to
COX-2 mRNA accumulation as pretreatment for 15 min with cycloheximide
(CHX, 1 µM) caused a superinduction of COX-2 mRNA accumulation in
unstimulated cells as well as in cells challenged with either TNF or
PMA. Expression of COX-2 protein in unstimulated MTAL cells was
attenuated by preincubation for 2 h with dexamethasone (Dex, 2 µM);
however, Dex had little or no effect on COX-2 expression in cells
challenged with either PMA or TNF. The time-dependent inhibition of
86Rb uptake by MTAL cells
challenged with TNF was diminished by pretreating cells with NS-398.
These data suggest that TNF-mediated induction of COX-2 protein
expression accounted for the lag-time required for this cytokine to
inhibit 86Rb uptake in MTAL cells.
tumor necrosis factor-; kidney; cyclooxygenase-2; prostaglandin
H synthase-2; medullary thick ascending limb; prostaglandins; cytokines
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