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Vol. 295, Issue 1, 417-422, October 2000

Evidence for Peroxynitrite Formation in Renal Ischemia-Reperfusion Injury: Studies with the Inducible Nitric Oxide Synthase Inhibitor L-N6-(1-Iminoethyl)lysine1

Lisa M. Walker, Patrick D. Walker, Syed Z. Imam, Syed F. Ali and Philip R. Mayeux

Departments of Pharmacology and Toxicology (L.M.W., P.R.M.), and Pathology (P.D.W.), University of Arkansas for Medical Sciences, Little Rock, Arkansas; and National Center for Toxicological Research, Division of Neurotoxicology, Jefferson, Arkansas (S.Z.I., S.F.A.)

Reactive oxygen species are suggested to participate in ischemia-reperfusion (I-R) injury. However, induction of inducible nitric oxide synthase (iNOS) and production of high levels of nitric oxide (NO) also contribute to this injury. NO can combine with superoxide to form the potent oxidant peroxynitrite (ONOO-). NO and ONOO- were investigated in a rat model of renal I-R injury using the selective iNOS inhibitor L-N6-(1-iminoethyl)lysine (L-NIL). Sprague-Dawley rats were subjected to 40 min of bilateral renal ischemia followed by 6 h of reperfusion with or without L-NIL administration. Control animals received a sham surgery and had plasma creatinine values of 0.4 ± 0.1 mg/dl. I-R surgery significantly increased plasma creatinine levels to 1.9 ± 0.3 mg/dl (P < .05) and caused renal cortical necrosis. L-NIL administration (3 mg/kg) in animals subjected to I-R significantly decreased plasma creatinine levels to 1.2 ± 0.10 mg/dl (P < .05 compared with I-R) and reduced tubular damage. ONOO- formation was evaluated by detecting 3-nitrotyrosine-protein adducts, a stable biomarker of ONOO- formation. Immunohistochemistry and HPLC revealed that the kidneys from I-R animals had increased levels of 3-nitrotyrosine-protein adducts compared with control animals. L-NIL-treated rats (3 mg/kg) subjected to I-R showed decreased levels of 3-nitrotyrosine-protein adducts. These results support the hypothesis that iNOS-generated NO mediates damage in I-R injury possibly through ONOO- formation.

1 This study was supported by an American Heart Association Heartland Affiliate Predoctoral Fellowship to L.M.W. and by National Institutes of Health Grant DK44716.

0022-3565/00/2951-0417$03.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 2000 by U.S. Government


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