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Alterations in Renal Endothelial Nitric Oxide Synthase Expression by Salt Diet in Angiotensin Type-1a Receptor Gene Knockout Mice

Keiko Sato^*, Minoru Kihara^*, Tatsuo Hashimoto^*, Kei Matsushita^*, Yu-Ichi Koide^*, Koichi Tamura^*, Nobuhito Hirawa^*, Yoshiyuki Toya^*, Akiyoshi Fukamizu and Satoshi Umemura^*

*Department of Second Internal Medicine, Yokohama City University, Yokohama; and the Center for Tsukuba Advanced Research Alliance, Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Japan.

Correspondence to Dr. Minoru Kihara, Department of Second Internal Medicine, Yokohama City University School of Medicine, Kanazawa-ku, Yokohama, 236-0004, Japan. Phone: 81-045-787-2635; Fax: 81-45-701-3738; E-mail: minoru{at}med.yokohama-cu.ac.jp

ABSTRACT. The effects of altered dietary salt intake and/or hydralazine-induced hypotension on renal endothelial nitric oxide synthase (eNOS) expression were determined in angiotensin type-1a receptor gene knockout (At1a–/–) and wild-type (At1a+/+) mice. In At1a–/– mice, the levels of renal cortical eNOS mRNA and protein were 5 times and 3.5 times higher, respectively, in the high-salt (4% NaCl) group than in the low-salt group (0.3% NaCl). Systemic BP of the high-salt group (105 ± 4.4 mmHg) was significantly higher than that of the low-salt group (77.0 ± 4.7 mmHg). When hydralazine was administered to the mutant mice fed a high-salt diet, BP was reduced to 72.5 ± 1.3 mmHg, with decreases in the levels of renal eNOS mRNA and protein expression to about half of those found in nontreated group. Consistent with the results for eNOS mRNA and protein expression, nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase activity and eNOS immunoreactivity localized in the endothelium of the renal vasculature changed parallel with the amount of salt intake. In contrast to mutant mice, At1a+/+ mice did not show any changes in renal eNOS expression during the manipulation of salt intake and/or hydralazine-induced hypotension. These results suggest that At1a receptor-mediated inputs play critical roles in maintaining renal vascular eNOS expression and activity during changes in salt-water balance and systemic BP.

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Copyright © 2008 by the American Society of Nephrology. Online ISSN: 1533-3450 Print ISSN: 1046-6673