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Am J Physiol Renal Physiol 282: F144-F150, 2002. First published August 21, 2001; doi:10.1152/ajprenal.0064.2001
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Vol. 282, Issue 1, F144-F150, January 2002

Role of cytochrome P-450 arachidonate metabolites in endothelin signaling in rat proximal tubule

Bruno A. Escalante1, John C. McGiff2, and Adebayo O. Oyekan3

1 Centro de Investigación y de Estudios Avanzados del Institúto Politécnico Nacional, Mexico DF 07300; 2 Department of Pharmacology, New York Medical College, Valhalla, New York 10595; and 3 College of Pharmacy and Health Sciences, Texas Southern University, Houston, Texas 77004

We examined the rat proximal tubule (PT) response to endothelin-1 (ET-1) in terms of 20-hydroxyeicosatetraenoic acid (HETE) dependency. Arachidonic acid (AA) (1 µM) decreased ouabain-sensitive 86Rb uptake from 2.1 ± 0.1 to 0.3 ± 0.08 ng Rb · 10 µg protein-1 · 2 min-1 (P < 0.05); 20-HETE (1 µM) had similar effects. Dibromododecenoic acid (DBDD) (2 µM), an inhibitor of omega -hydroxylase, abolished the inhibitory action of AA on 86Rb uptake whereas the PT response to 20-HETE was unaffected. ET-1 at 0.1, 1, 10, and 100 nM reduced 86Rb uptake from 2.8 ± 0.3 in control PTs to 2.4 ± 0.2, 1.7 ± 0.1, 0.67 ± 0.08, and 0.1 ± 0.03 ng Rb · 10 µg protein-1 · 2 min-1, respectively. DBDD (2 µM) abolished the inhibitory effect of ET-1 on 86Rb uptake as did BMS182874 (1 µM), an ETA-selective receptor antagonist. ET-1 (100 nM) significantly increased PT 20-HETE release by ~50%, an effect prevented by DBDD. Nomega -nitro-L-arginine-methyl ester (L-NAME), given for 4 days to inhibit nitric oxide synthase (NOS), increased arterial pressure from 92 ± 12 to 140 ± 8 mmHg and increased endogenous release of 20-HETE from isolated PTs (measured by gas chromatography/mass spectrometry). In L-NAME-treated PTs, but not in control PTs, 0.1 µM AA inhibited ouabain-sensitive 86Rb uptake by >40%; the response to AA was attenuated by DBDD. We conclude that, in the PTs, 1) 20-HETE is a second messenger for ET-1 and 2) conversion of AA to 20-HETE is augmented when NOS is inhibited.

20-hydroxyeicosatetraenoic acid; arachidonic acid metabolites; endothelin-1; nitric oxide


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