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Departments of 1 Internal Medicine and 2 Physiology, University of Michigan, Ann Arbor, Michigan 48104; and 3 National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892
Renal medullary
prostaglandins are believed to exert an important functional role in
antagonizing vasopressin effects in dehydration. Studies were
undertaken to determine the effect of hyperosmolality on cyclooxygenase
(COX) isoform expression in the renal medulla. COX-1 and
COX-2 mRNA and protein levels were determined by RT-PCR or Western
blotting in Sprague-Dawley rats on varying water intakes, in
Brattleboro rats and in Long-Evans controls. Over a wide range of
urinary tonicity, COX-2 expression correlated closely with urine
osmolality levels (R = 0.872). COX-1
levels did not vary. Immunolocalization showed that the stimulation of
COX-2 expression by dehydration occurred predominantly in the
collecting duct. Hypertonicity caused by addition of NaCl produced a
dose- and time-dependent stimulation of COX-2 expression in mIMCD-K2
cells as well as in MDCK cells. COX-1 was unaffected. In the same cell lines, mannitol, sucrose, and raffinose also had a stimulatory effect.
The tonicity-stimulated COX-2 expression in mIMCD-K2 cells was almost
completely blocked by a tyrosine kinase inhibitor, genistein at 100 µM. In MDCK cells transfected with a 2.7-kb COX-2 promoter and
lacZ reporter construct, NaCl induced a twofold increase in
-galactosidase activity. Using mIMCD-K2 cells, hypertonic NaCl (600 mosmol/kgH2O for 24 h) induced a
33-fold increase in PGE2 release determined by enzyme
immunoassay, an effect completely blocked by 3 µM indomethacin or the
COX-2-specific blocker
N-(2-cyclohexy-4-nitrophenyl)methanesulfonamide (NS-398). We conclude that in inner medulla, COX-2 but not
COX-1 is upregulated by hyperosmolality.
prostaglandin H synthase; inner medulla; hyperosmolality; prostaglandins; sodium chloride; promoter
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