Preferences help
enabled [disable] Abstract
Number of results
2004 | 51 | 4 | 933-942
Article title

Nitric oxide - superoxide cooperation in the regulation of renal Na+,K+-ATPase.

Title variants
Languages of publication
The aim of this study was to investigate whether endogenous superoxide anion is involved in the regulation of renal Na+,K+-ATPase and ouabain-sensitive H+,K+-ATPase activities. The study was performed in male Wistar rats. Compounds modulating superoxide anion concentration were infused under general anaesthesia into the abdominal aorta proximally to the renal arteries. The activity of ATPases was assayed in isolated microsomal fraction. We found that infusion of a superoxide anion-generating mixture, xanthine oxidase (1 mU/min per kg) + hypoxanthine (0.2 μmol/min per kg), increased the medullary Na+,K+-ATPase activity by 49.5% but had no effect on cortical Na+,K+-ATPase and either cortical or medullary ouabain-sensitive H+,K+-ATPase. This effect was reproduced by elevating endogenous superoxide anion with a superoxide dismutase inhibitor, diethylthiocarbamate. In contrast, a superoxide dismutase mimetic, TEMPOL, decreased the medullary Na+,K+-ATPase activity. The inhibitory effect of TEMPOL was abolished by inhibitors of nitric oxide synthase (L-NAME), soluble guanylate cyclase (ODQ) and protein kinase G (KT5823). The stimulatory effect of diethylthiocarbamate was not observed in animals pretreated with a synthetic cGMP analogue, 8-bromo-cGMP. An inhibitor of NAD(P)H oxidase, apocynin (1 μmol/min per kg), decreased the Na+,K+-ATPase activity in the renal medulla and its effect was prevented by L-NAME, ODQ or KT5823. In contrast, a xanthine oxidase inhibitor, oxypurinol, administered at the same dose was without effect. These data suggest that NAD(P)H oxidase-derived superoxide anion increases Na+,K+-ATPase activity in the renal medulla by reducing the availability of NO. Excessive intrarenal generation of superoxide anion may upregulate medullary Na+,K+-ATPase leading to sodium retention and blood pressure elevation.

Physical description
  • Department of Pathophysiology, Medical University, Lublin, Poland
  • Department of Pathophysiology, Medical University, Lublin, Poland
  • Department of Pathophysiology, Medical University, Lublin, Poland
  • Department of Pathophysiology, Medical University, Lublin, Poland
  • Aperia A, Holtback U, Syren ML, Svensson LB, Fryckstedt J, Greengard P. (1994) Activation/deactivation of renal Na+,K+-ATPase: a final common pathway for regulation of natriuresis. FASEB J.; 8: 436-9.
  • BeŁtowski J, Wójcicka G. (2002) Spectrophotometric method for the determination of renal ouabain-sensitive H+,K+-ATPase activity. Acta Biochim Polon.; 49: 515-27.
  • BeŁtowski J, Marciniak A, Wójcicka G, Górny D. (2003) Nitric oxide decreases renal medullary Na+,K+-ATPase activity through cyclic GMP-protein kinase G dependent mechanism. J Physiol Pharmacol.; 54: 191-210.
  • Chabrashvili T, Tojo A, Onozato ML, Kitiyakara C, Quinn MT, Fujita T, Welch WJ, Wilcox CS. (2002) Expression and cellular localization of classic NADPH oxidase subunits in the spontaneously hypertensive rat kidney. Hypertension.; 39: 269-74.
  • Cowley AW, Mori T, Mattson D, Zou AP. (2003) Role of renal NO production in the regulation of medullary blood flow. Am J Physiol Regul Integr Comp Physiol.; 284: R1355-69.
  • Féraille E, Doucet A. (2001) Sodium-potassium- adenosinetriphosphatase-dependent sodium transport in the kidney: hormonal control. Physiol Rev.; 81: 345-418.
  • Garvin JL, Ortiz PA. (2003) The role of reactive oxygen species in the regulation of tubular function. Acta Physiol Scand.; 179: 225-32.
  • Geiszt M, Kopp JB, Várnai P, Leto TL. (2000) Identification of Renox, an NAD(P)H oxidase in kidney. Proc Natl Acad Sci USA.; 97: 8010-4.
  • Guzman NJ, Fang MZ, Tang SS, Ingelfinger JR, Garg LC. (1995) Autocrine inhibition of Na,K-ATPase by nitric oxide in mouse proximal tubule epithelial cells. J Clin Invest.; 95: 2083-8.
  • Haque MZ, Majid DS. (2004) Assessment of renal functional phenotype in mice lacking gp91PHOX subunit of NAD(P)H oxidase. Hypertension.; 43: 335-40.
  • Hoagland KM, Maier KG, Roman RJ. (2003) Contributions of 20-HETE to the antihypertensive effects of Tempol in Dahl salt-sensitive rats. Hypertension.; 41: 697-702.
  • Hurst RO. (1964) The determination of nucleotide phosphorus with a stannous chloride-hydrazine sulphate reagent. Can J Biochem.; 42: 287-92.
  • Kang DG, Kim JW, Lee J. (2000) Effects of nitric oxide synthesis inhibition on the Na,K-ATPase activity in the kidney. Pharmacol Res.; 41: 121-5.
  • Kiroytscheva M, Cheval L, Carranza ML, Martin PY, Favre H, Doucet A, Féraille E. (1999) Effect of cAMP on the activity and the phosphorylation of Na+,K+-ATPase in rat thick ascending limb of Henle. Kidney Int.; 55: 1819-31.
  • Kone BC, Higham S. (1999) Nitric oxide inhibits transcription of the Na+-K+-ATPase α1-subunit gene in an MTAL cell line. Am J Physiol.; 276: F614-21.
  • Laycock SK, Vogel T, Forfia PR, Tuzman J, Xu X, Ochoa M, Thompson CI, Nasjletti A, Hintze TH. (1998) Role of nitric oxide in the control of renal oxygen consumption and the regulation of chemical work in the kidney. Circ Res.; 82: 1263-71.
  • Li N, Yi FX, Spurrier JL, Bobrowitz CA, Zou AP. (2002) Production of superoxide through NADH oxidase in thick ascending limb of Henle's loop in rat kidney. Am J Physiol Renal Physiol..; 282: F1111-9.
  • Lowry OH, Rosebrough NI, Farr AL, Randall RJ. (1951) Protein measurement with the Folin phenol reagent. J Biol Chem.; 193: 265-75.
  • Majid DS, Nishiyama A. (2002) Nitric oxide blockade enhances renal responses to superoxide dismutase inhibition in dogs. Hypertension.; 39: 293-7.
  • Makino A, Skelton MM, Zou AP, Roman RJ, Cowley AW. (2002) Increased renal medullary oxidative stress produces hypertension. Hypertension.; 39: 667-72.
  • McGiff JC, Quilley J. (1999) 20-HETE and the kidney: resolution of old problems and new beginnings. Am J Physiol.; 277: R607-23.
  • Meng S, Roberts LJ, Cason GW, Curry TS, Manning RD. (2002) Superoxide dismutase and oxidative stress in Dahl salt-sensitive and -resistant rats. Am J Physiol Regul Integr Comp Physiol.; 283: R732-8.
  • Ortiz PA, Garvin JL. (2002a) Interaction of O2- and NO in the thick ascending limb. Hypertension.; 39: 591-6.
  • Ortiz PA, Garvin JL. (2002b) Role of nitric oxide in the regulation of nephron transport. Am J Physiol Renal Physiol.; 282: F777-84.
  • Ortiz PA, Garvin JL. (2002c) Superoxide stimulates NaCl absorption by the thick ascending limb. Am J Physiol Renal Physiol..; 283: F957-62.
  • Racasan S, Turkstra E, Joles JA, Koomans HA, Braam B. (2003) Hypoxanthine plus xanthine oxidase causes profound natriuresis without affecting renal blood flow autoregulation. Kidney Int.; 64: 226-31.
  • Ren Y, Carretero OA, Garvin JL. (2002) Mechanism by which superoxide potentiates tubuloglomerular feedback. Hypertension.; 39: 624-8.
  • Rodrigo R, Trujillo S, Bosco C, Orellana M, Thielemann L, Araya J. (2002) Changes in (Na+ K)-adenosine triphosphatase activity and ultrastructure of lung and kidney associated with oxidative stress induced by acute ethanol intoxication. Chest.; 121: 589-96.
  • Scavone C, Scanlon C, McKee M, Nathanson JA. (1995) Atrial natriuretic peptide modulates sodium and potassium-activated adenosine triphosphatase through a mechanism involving cyclic GMP and cyclic GMP-dependent protein kinase. J Pharmacol Exp Ther.; 272: 1036-43.
  • Schnackenberg CG. (2002a) Oxygen radicals in cardiovascular-renal disease. Curr Opin Pharmacol.; 2: 121-5.
  • Schnackenberg CG. (2002b) Physiological and pathophysiological roles of oxygen radicals in the renal microvasculature. Am J Physiol Regul Integr Comp Physiol.; 282: R335-42.
  • Schoonmaker GC, Fallet RW, Carmines PK. (2000) Superoxide anion curbs nitric oxide modulation of afferent arteriolar ANG II responsiveness in diabetes mellitus. Am J Physiol Renal Physiol.; 278: F302-9.
  • Shiose A, Kuroda J, Tsuruya K, Hirai M, Hirakata H, Naito S, Hattori M, Sakaki Y, Sumimoto H. (2001) A novel superoxide-producing NAD(P)H oxidase in kidney. J Biol Chem.; 276: 1417-23.
  • Shokoji T, Fujisawa Y, Kimura S, Rahman M, Kiyomoto H, Matsubara K, Moriwaki K, Aki Y, Miyatake A, Kohno M, Abe Y, Nishiyama A. (2004) Effects of local administrations of tempol and diethyldithio-carbamic on peripheral nerve activity. Hypertension.; 44: 236-43
  • Varela M, Herrera M, Garvin JL. (2004) Inhibition of Na-K-ATPase in thick ascending limbs by NO depends on O2- and is diminished by a high-salt diet. Am J Physiol Renal Physiol.; 287: F224-30.
  • Welch WJ, Tojo A, Wilcox CS. (2000) Roles of NO and oxygen radicals in tubuloglomerular feedback in SHR. Am J Physiol Renal Physiol.; 278: F769-76.
  • Zhang C, Mayeux PR. (2001) NO-cGMP signaling modulates regulation of Na+,K+-ATPase activity by angiotensin II in rat proximal tubules. Am J Physiol Renal Physiol.; 280: F474-9.
  • Zhang C, Imam SZ, Ali SF, Mayeux PR. (2002) Peroxynitrite and the regulation of Na+,K+-ATPase activity by angiotensin II in the rat proximal tubule. Nitric Oxide.; 7: 30-5.
  • Zou AP, Cowley AW. (2003) Reactive oxygen species and molecular regulation of renal oxygenation. Acta Physiol Scand.; 179: 233-41.
  • Zou AP, Li N, Cowley AW. (2001) Production and actions of superoxide in the renal medulla. Hypertension.; 37: 547-53.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.