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Am J Physiol Renal Physiol 288: F182-F187, 2005. First published September 14, 2004; doi:10.1152/ajprenal.00036.2004
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Nitrite-derived nitric oxide formation following ischemia-reperfusion injury in kidney

Masumi Okamoto,1,2,* Koichiro Tsuchiya,3 Yasuhisa Kanematsu,1 Yuki Izawa,1 Masanori Yoshizumi,1 Susumu Kagawa,2 and Toshiaki Tamaki1,*

Departments of 1Pharmacology and 2Urology, University of Tokushima School of Medicine, and 3Department of Clinical Pharmacology, Graduate School of Pharmaceutical Sciences, University of Tokushima, Tokushima, Japan

Submitted 4 February 2004 ; accepted in final form 9 September 2004

Nitric oxide (NO) is synthesized from L-arginine by nitric oxide synthase (NOS), and nitrite and nitrate are believed to be waste forms of NO. We previously reported an enzyme-independent pathway of NO generation from nitrite in acidic conditions. In this study, we show nitrite-derived NO formation in renal ischemia-reperfusion injury using electron paramagnetic resonance (EPR) spectroscopy. In this experiment, we utilized a stable isotope of [15N]nitrite as a source of nitrite to distinguish L-arginine-derived NO from [15N]nitrite-derived 15NO. Intravenous infusion of a stable isotope of [15N]nitrite (15NO2–) facilitated the formation of Hb15NO during renal ischemia, which demonstrated that the origin of NO was nitrite. The EPR signal of Hb15NO in kidney appeared after 40 min of renal ischemia, and renal reperfusion decreased the Hb15NO level in the kidney and increased it in blood by contrast. In addition, the amount of HbNO was nitrite concentration dependent, and this formation was NOS independent. Our findings suggest that nitrite can be an alternative source of NO in ischemic kidney and that it binds with hemoglobin and then is spread by the circulation after reperfusion.

HbNO; electron paramagnetic resonance; NG-nitro-L-arginine methyl ester


Address for reprint requests and other correspondence: T. Tamaki, Dept. of Pharmacology, Univ. of Tokushima School of Medicine, 3–18-15 Kuramoto, Tokushima 770-8503, Japan (E-mail: tamaki{at}basic.med.tokushima-u.ac.jp)




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