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Importance of renal mitochondria in the reduction of TEMPOL, a nitroxide radical

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Abstract

Spin probing methods using an electron spin resonance (ESR) spectrometer are used extensively and bring us a lot of information about in vivo redox mechanisms. However, the in vivo reducing mechanisms of exogenous nitroxide radicals, which serve as typical spin probing reagents are not clear. To clarify this, we examined the sequential kinetics of a spin probe, 4-hydroxy 2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL) in the in vivo organs, tissue homogenates and subcellular fractions of kidney and liver using an in vivo and X-band ESR spectrometers. As a parameter of reducing activity, we calculated the half-life of TEMPOL from the decay curve of ESR signal intensity. The half-life of TEMPOL in the whole organs and homogenates of the kidney was significantly shorter than that of the liver, this indicates that the kidney has more reducing activity against TEMPOL as compared to the liver. Subcellular fractional studies revealed that this reducing activity of the kidney mainly exists in the mitochondria. Contrarily, in addition to reduction in the mitochondria, TEMPOL in the liver was reduced by the microsome and cytosol.

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References

  1. Rosen GM, Rauckman EJ: Formation and reduction of a nitroxide radicals by liver microsomes. Biochem Pharmacol 26: 675-678, 1977

    Google Scholar 

  2. Iannone A, Bini A, Swartz HM, Tomasi A, Vannini V: Metabolism in rat liver microsomes of the nitroxide spin probe tempol. Biochem Pharmacol 38: 2581-2586, 1989

    Google Scholar 

  3. Iannone A, Tomasi A, Vannini V, Swartz HM: Metabolism of nitroxide spin labels in subcellular fraction of rat liver I. Reduction by microsomes. Biochim Biophys Acta 1034: 285-289, 1990

    Google Scholar 

  4. Miura Y, Utsumi H, Kashiwagi M, Hamada A: Effects of oxygen on the membrane structure and the metabolism of lipophilic nitroxide in rat liver microsomes. J Biochem 108: 516-518, 1990

    Google Scholar 

  5. Quintanilha AT, Packer L: Surface localization of sites of reduction of nitroxide spin-labeled molecules in mitochondria. Proc Natl Acad Sci USA 74: 570-574, 1977

    Google Scholar 

  6. Chen K, Morse PD II, Swartz HM: Kinetics of enzyme-mediated reduction of lipid soluble nitroxide spin labels by living cells. Biochim Biophys Acta 943: 477-484, 1988

    Google Scholar 

  7. Chen K, Glockner JF, Morse PD II, Swartz HM: Effects of oxygen on the metabolism of nitroxide spin labels in cells. Biochemistry 28: 2496-2501, 1989

    Google Scholar 

  8. Couet WR, Brasch RC, Sosnovsky G, Tozer TN: Factors affecting nitroxide reduction in ascorbate solution and tissue homogenates. Magn Reson Imag 3: 83-88, 1985

    Google Scholar 

  9. Giotta GJ, Wang HH: Reduction of nitroxide free radicals in biological materials. Biochem Biophys Res Commun 46: 1576-1580, 1972

    Google Scholar 

  10. Eriksson UG, Brasch RC, Tozer TN: Non-enzymatic bioreduction in rat liver and kidney of nitroxyl spin labels, potential contrast agents in magnetic resonance imaging. Drug Metab Disp 15: 155-160, 1987

    Google Scholar 

  11. Iannone A, Tomasi A, Vannini V, Swartz HM, Metabolism of nitroxide spin labels in subcellular fraction of rat liver II. Reduction in the cytosol. Biochim Biophys Acta 1034: 290-293, 1990

    Google Scholar 

  12. Swartz HM, Sentjurc M, Morsell PD: Cellular metabolism of water-soluble nitroxides: Effect on rate of reduction of cell/nitroxide ratio, oxygen concentrations and permeability of nitroxides. Biochim Biophys Acta 888: 82-90, 1986

    Google Scholar 

  13. Couet WR, Eriksson UG, Tozer TN, Tuck LD, Wesbey GE, Nitecki D, Brasch RC: Pharmacokinetics and metabolic fate of two nitroxides potentially useful as contrast agents for magnetic resonance imaging. Pharm Res 1: 203-209, 1984

    Google Scholar 

  14. Inaba K, Nakashima T, Shima T, Mitsuyoshi H, Sakamoto Y, Okanoue T, Kashima K, Hashiba M, Nishikawa H, Watari H: Hepatic damage influences the decay of nitroxide radicals in mice — an in vivo ESR study. Free Radic Res 27: 37-43, 1997

    Google Scholar 

  15. Togashi H, Shinzawa H, Ogata T, Matsuo T, Ohno S, Saito K, Yamada N, Yokoyama H, Noda H, Oikawa K, Kamada H, Takahashi T: Spatiotemporal measurement of free radical elimination in the abdomen using an in vivo ESR-CT imaging system. Free Radic Biol Med 25: 1-8, 1998

    Google Scholar 

  16. Fillit H, Elion E, Sullivan J, Sherman R, Zabriskie JB: Thiobarbituric acid reactive material in uremic blood. Nephron 29: 40-43, 1981

    Google Scholar 

  17. Dasgupta A, Hussain S, Ahmad S: Increased lipid peroxidation in patients on maintenance hemodialysis. Nephron 60: 56-59, 1992

    Google Scholar 

  18. Gotoh M, Nagase S, Aoyagi K, Hirayama A, Takemura K, Ueda A, Tomoda C, Kikuchi H, Koyama A: Thiobarbituric acid reactive substances are increased in the subcutaneous fat tissue of patients with end-stage renal disease. Nephrol Dial Transplant 12: 713-717, 1997

    Google Scholar 

  19. Nagase S, Aoyagi K, Hirayama A, Gotoh M, Ueda A, Tomida C, Kamezaki T, Nagai Y, Kikuchi H, Koyama A: Favorable effects of hemodialysis on decreased serum antioxidant activity in hemodialysis patients demonstrated by electron spin resonance. J Am Soc Nephrol 8: 1157-1163, 1997

    Google Scholar 

  20. Hirata H, Iwai H, Ono M: Analysis of a flexible surface-coil-type resonator for magnetic resonance measurements. Rev Sci Instrum 66: 4529-4534, 1995

    Google Scholar 

  21. Tada M, Yokoyama H, Toyoda Y, Ohya H, Ito T, Ogata T: Surface-coiltype resonators for in vivo temporal ESR measurements in different organs of nitroxide-treated rats. Appl Magn Reson 18: 575-582, 2000

    Google Scholar 

  22. Utsumi H, Muto E, Hamada A: In vivo ESR measurement of free radicals in whole mice. Biochem Biophys Res Commun 172: 1342-1348, 1990

    Google Scholar 

  23. Takeshita K, Hamada A, Utsumi H: Mechanisms related to reduction of radical in mouse lung using an L-band ESR spectrometer. Free Radic Biol Med 26: 951-960, 1999

    Google Scholar 

  24. Ishida S, Matsumoto S, Yokoyama H, Mori N, Kumashiro H, Tsuchihashi N, Ogata T, Yamada M, Ono M, Kitajima T, Kamada H, Yoshida H: An ESR-CT imaging of the head of a living rat receiving an administration of a nitroxide radical. Magn Reson Imag 10: 21-27, 1992

    Google Scholar 

  25. Lin Y, Yokoyama H, Ishida S, Tsuchihashi N, Ogata T: In vivo electron spin resonance analysis of nitroxide radicals injected into a rat by a flexible surface-coil-type resonator as an endoscope-or a stetho-scope-like device. Magn Reson Mater Phy 2: 99-104, 1997

    Google Scholar 

  26. Yokoyama H, Ogata T, Tsuchihashi N, Hiramatsu M, Mori N: A spatiotemporal study on the distribution of intraperitoneally injected nitroxide radical in the rat head using an in vivo ESR imaging system. Magn Reson Imag 10: 21-27, 1992

    Google Scholar 

  27. Takeshita K, Utsumi H, Hamada A: ESR measurement of radical clearance in lung of whole mouse. Biochem Biophys Res Commun 177: 874-880, 1991

    Google Scholar 

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Authors and Affiliations

  1. Department of Internal Medicine, Institute of Clinical Medicine, University of Tsukuba, Ibaraki, Japan

    Atsushi Ueda, Sohji Nagase, Aki Hirayama & Akio Koyama

  2. Institute for Life Support Technology and Regional Joint Research Project of Yamagata Prefecture, Yamagata Public Corporation for the Development of Industry, Yamagata, Japan

    Hidekatsu Yokoyama, Mika Tada, Hiroyuki Noda, Horoaki Ohya & Hitoshi Kamada

Authors
  1. Atsushi Ueda
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  2. Sohji Nagase
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  3. Hidekatsu Yokoyama
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  4. Mika Tada
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  5. Hiroyuki Noda
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  6. Horoaki Ohya
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  7. Hitoshi Kamada
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  8. Aki Hirayama
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  9. Akio Koyama
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Ueda, A., Nagase, S., Yokoyama, H. et al. Importance of renal mitochondria in the reduction of TEMPOL, a nitroxide radical. Mol Cell Biochem 244, 119–124 (2003). https://doi.org/10.1023/A:1022477530291

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  • DOI: https://doi.org/10.1023/A:1022477530291

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