This study was designed to characterize the time course of the effects of dexamethasone (2.5 mg kg-1 week-1, sc) on the renal arachidonate-prostaglandin (PG) system and to define the effect of the steroid on the interstitial cells of the renal inner medulla (RIC). The RIC are rich in triglycerides, which, due to their high content of arachidonic acid, may be a source of arachidonate for PG synthesis during conditions of phospholipase inhibition. After 1 day of dexamethasone treatment, the urinary excretion of PGE2 and PGF2 alpha was reduced to about 50% of the control value (P less than 0.05), and angiotensin II-induced release of arachidonic acid and PGs from renal medulla slices was blunted (P less than 0.05). In contrast, dexamethasone treatment did not affect ionophore A23187-induced release of PGs and arachidonic acid from renal medulla slices. By day 3 of dexamethasone treatment, urinary excretion of PGE2 and PGF2 alpha had returned to control levels, and by days 5 and 14 the excretion rates of both were clearly increased (P less than 0.05). The rise in urinary PG excretion was accompanied by a reduction of renal 15-hydroxyprostaglandin dehydrogenase activity, augmentation of renal medulla microsomal PG synthetase activity, and diminution of renal medulla triglycerides, the latter associated with a reduction in the number of RIC and of RIC osmiophilic granules. This study demonstrates that the effects of dexamethasone on urinary PG are biphasic; this may reflect the changing balance between the opposing actions of the steroid on renal PG-synthesizing and catabolizing enzymes and the inhibitory effect on the synthesis of renal PG that is linked to activation of specific renal lipases by endogenous factors such as angiotensin II.