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1 Division of Nephrology and Hypertension, Georgetown University, Washington, DC, USA
* To whom correspondence should be addressed. E-mail: welchw{at}georgetown.edu.
Renal oxygen tension is substantially lower in the medulla than in the cortex and is reduced in hypertensive rats, a model of oxidative stress. Expression of NADPH oxidase, the primary source for superoxide (O2 .-) in the kidney is elevated in hypertension. Since molecular oxygen (O2) is required for O2 .-) formation, we tested the hypothesis that renal NADPH oxidase activity is limited by low O2. Superoxide production by rat kidney tissue or cultured cells exposed to levels of PO2 that mimics those in the kidney was assessed by lucigenin-enhanced chemiluminescence. NADPH-dependent O2 .-) production by kidney homogenates decreased reversibly by 60-90% after graded reductions of ambient O2 from 10 to 0% (76 to 2 mmHg PO2). The NADPH-dependent O2 .-) production by the kidney homogenate was reduced by decreasing PO2 below ~30 mmHg. The response of tissue homogenates to low PO2 was not different between normotensive and hypertensive rats. Similarly, NADPH-dependent O2 .-) production was lower during 2% O2 compared to 10% O2 in rat proximal tubule cells (-57±1%), VSM (-42±5%), cardiomyocytes (-57±1%) and mouse inner medulla collecting duct cells (-58±3%). We conclude that superoxide production by NADPH oxidase is dependent on availability of O2. Therefore superoxide generation may be limited in the kidney, both in the normal renal medulla and in the cortex of hypertensive and diabetic kidneys.
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