on January 12, 2004
Published online before print January 12, 2004, doi: 10.1161/01.HYP.0000113295.31481.36
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Submitted on September 29, 2003
From the Department of Physiology, Medical College of Wisconsin, Milwaukee. * To whom correspondence should be addressed. E-mail: tmori{at}mcw.edu.
Abstract--The effects of NaCl, glucose, and thyroid hormone on the production of superoxide (O2·-) within the renal medulla of Sprague-Dawley rats were examined. Responses of intracellular superoxide [O2·-]i in isolated medullary thick ascending limbs (mTALs) were studied using real-time fluorescent microscopy with measurement of the dehydroethidium (DHE) to ethidium (Eth) conversion ratio (Eth/DHE ratio unit). The results demonstrated that elevations of extracellular NaCl (from 152 to 252 mmol/L), D-glucose (from 5 to 25 mmol/L), and triiodo-thyronine (T3; 10 µmol/L) significantly increased [O2·-]i levels. Preincubation with superoxide scavenger 4,5-dihydroxy-1,3-benzene-disulfonic acid (1 mmol/L) significantly inhibited these responses. Stimulation with equamolar amounts of choline chloride or L-glucose failed to increase [O2·-]i, indicating that these O2·- responses were not determined by changes in osmolality. The responses to NaCl, D-glucose, and T3 were abolished by pretreatment with the Na+/K+-ATPase pump inhibitor ouabain (4 mmol/L) and with Na+/H+ -exchanger inhibitor dimethylamiloride (100 µmol/L). We conclude that elevations of extracellular NaCl, D-glucose, or T3 levels can activate both the Na+/K+-ATPase pump and Na+/H+ exchanger in mTAL, which, in turn, is associated with increased intracellular concentrations of superoxide.
Revised on October 27, 2003
Renal Oxidative Stress in Medullary Thick Ascending Limbs Produced by Elevated NaCl and Glucose
Takefumi Mori* and Allen W. Cowley Jr.
Key words: oxidative stress • Na+, K+-transporting ATPase • sodium pump • antiporters • diabetes mellitus • hypertension, sodium-dependent • fluorescence
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