on September 15, 2003
Published online before print September 15, 2003, doi: 10.1161/01.HYP.0000091821.39824.09
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Submitted on February 20, 2003
From the Department of Physiology, Medical College of Wisconsin, Milwaukee. * To whom correspondence should be addressed. E-mail: cowley{at}mcw.edu.
Abstract--The source of superoxide (O2·-) production and cell-to-cell interactions of O2·- and nitric oxide (NO) in response to angiotensin II (AngII) were studied by fluorescence microscopic techniques to image rat renal outer medullary microtissue strips. Changes in intracellular O2·- were determined by dihydroethidium-ethidium ratios, and NO was determined with 4,5-diaminofluorescein diacetate. AngII (1 µmol/L) significantly increased O2·- in the isolated, medullary thick ascending limb (mTAL). These responses were inhibited by the superoxide dismutase mimetic 4-hydroxytetramethylpiperidine-1-oxyl (TEMPOL) and by the NAD(P)H oxidase inhibitors diphenylene iodonium and apocynin. AngII did not increase O2·- in either pericytes of isolated, intact vasa recta (VR) or pericytes of VR with a disrupted endothelium, even when surrounded by mTAL. However, AngII did increase O2·- when the tissue strips were preincubated with the NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO), indicating that cross-talk of O2·- from mTAL to the VR occurred but was normally inhibited by NO. Also, tissue O2·- reduction by TEMPOL increased the diffusion of NO from mTAL to the pericytes, indicating that cross-talk of NO from the mTAL to the VR is also inhibited by O2·-. We conclude that AngII stimulates O2·- production in mTAL via the NAD(P)H oxidase pathway and that interactions of O2·- and NO ultimately determine the effectiveness of in situ free-radical cross-talk between the mTAL and the VR.
Revised on March 12, 2003
Angiotensin II-NAD(P)H Oxidase-Stimulated Superoxide Modifies Tubulovascular Nitric Oxide Cross-Talk in Renal Outer Medulla
Takefumi Mori and Allen W. Cowley Jr*
Key words: free radicals • rats • oxidative stress • renal blood flow • angiotensin II
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