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Vol. 101, No. 2, 2005
Issue release date: October 2005

Oxidative Stress Decreases Klotho Expression in a Mouse Kidney Cell Line

Mitobe M. · Yoshida T. · Sugiura H. · Shirota S. · Tsuchiya K. · Nihei H.
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Background/Aims: Defects in klotho gene expression in the mouse result in a syndrome that resembles human aging. We recently identified expression of klotho in a mouse inner medullary collecting duct (mIMCD3) cell line for the first time, and in the present study we explored the physiological relevance of the regulation of klotho expression in the presence of oxidant stress injury. Methods: Klotho expression was analyzed by real-time PCR, Western blot, and immuocytochemical staining during exposure to hydrogen peroxide (H2O2). Overexpression of the klotho gene was induced by klotho adenoviruses, and the number of apoptotic cells was counted by flowcytometry. Results: Oxidant stress injury by H2O2 dose-dependently reduced klotho expression and diminished klotho staining. There were fewer apoptotic cells among the klotho-transfected cells than among the control cells. Conclusion: Klotho is expressed in cell line mIMCD3, and the klotho gene may be involved in the process of oxidative stress injury and apoptosis in this cell line.

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  1. Kuro-o M, Matsumura Y, Aizawa H, Kawaguchi H, Suga T, Utsugi T, Ohyama Y, Kurabayashi M, Kaname T, Kume E, Iwasaki H, Iida A, Shiraki-Iida T, Nishikawa S, Nagai R, Nabeshima Y: Mutation of the mouse klotho gene leads to a syndrome resembling ageing. Nature 1997;390:45–51.
  2. Matsumura Y, Aizawa H, Shiraki-Iida T, Nagai R, Kuro-o M, Nabeshima Y: Identification of the human klotho gene and its two transcripts encoding membrane and secreted klotho protein. Biochem Biophys Res Commun 1998;242:626–630.
  3. Saito Y, Yamagishi T, Nakamura T, Ohyama Y, Aizawa H, Suga T, Matsumura Y, Masuda H, Kurabayashi M, Kuro-o M, Nabeshima Y, Nagai R: Klotho protein protects against endothelial dysfunction. Biochem Biophys Res Commun 1998;248:324–329.
  4. Shiraki-Iida T, Aizawa H, Matsumura Y: Structure of the mouse klotho gene and its two transcripts encoding membrane and secreted protein. FEBS Lett 1998;424:6–10.
  5. Aizawa H, Saito Y, Nakamura T, Inoue M, Imanari T, Ohyama Y, Matsumura Y, Masuda H, Oba S, Mise N, Kimura K, Hasegawa A, Kurabayashi M, Kuro-o M, Nabeshima Y, Nagai R: Downregulation of the klotho gene in the kidney under sustained circulatory stress in rats. Biochem Biophys Res Commun 1998;249:865–871.
  6. Ohyama Y, Kurabayashi M, Matsuda H, Nakamura T, Aihara Y, Kaname T, Suga T, Arai M, Aizawa H, Matsumura Y, Kuro-o M, Nabeshima Y: Molecular cloning of rat klotho cDNA: markedly decreased expression of klotho by acute inflammatory stress. Biochem Biophys Res Commun 1998;251:920–925.
  7. Koh N, Fujimori T, Nishiguchi S, Tamori A, Shiomi S, Nakatani T, Sugimura K, Kishimoto T, Knoshita S, Kroki T, Nabeshima Y: Severely reduced production of klotho in human chronic renal failure kidney. Biochem Biophys Res Commun 2001;280:1015–1020.
  8. Kelly KJ, Molitoris BA: Acute renal failure in the new millennium: Time to consider combination therapy. Semin Nephrol 2000;20:4–19.
  9. Nath KA, Norby SM: Reactive oxygen species and acute renal failure. Am J Med 2000;109:655–678.

    External Resources

  10. Yu BU: Cellular defenses against damage from reactive oxygen species. Physiol Rev 1994;74:139–162.
  11. Lloberas N, Torras J, Herrero-Fresneda I, Cruzado JM, Riera M, Hurtado I, Grinyo JM: Postischemic renal oxidative stress induces inflammatory response through PAF and oxidized phospholipids: Prevention by antioxidant treatment. FASEB J 2002;16:908–910.
  12. Shumer M, Colombel MC, Sawczuk IS, Gobe G, Connor J, O’Toole KM, Olsson CA, Wise G.J, Buttyan R: Morphologic, biochemical, and molecular evidence of apoptosis during the reperfusion phase after brief periods of renal ischemia. Am J Pathol 1992;140:831–838.

    External Resources

  13. Chien CT, Lee OH, Chen CF, Ma MC, Lai MK, Hsu SM: De novo demonstration and co-localization of free-radical production and apoptosis formation in rat kidney subjected to ischemia/reperfusion. J Am Soc Nephrol 2001;12:973–982.
  14. Padanilam BJ: Cell death induced by acute renal injury: A perspective on the contributions of apoptosis and necrosis. Am J Physiol 2003;284:F608–F627.
  15. Saikumar P, Venkatachalam M.A: Role of apoptosis in hypoxic/ischemic damage in the kidney. Semin Nephrol 2003;23:511–521.
  16. Rauchman MI, Nigam SK, Delpire E, Gullans SR: An osmotically tolerant inner medullary collecting duct cell line from an SV40 transgenic mouse. Am J Physiol 1993;265:F416–F424.
  17. Yoshida T, Tang SS, Hsiao LL, Jensen RV, Ingelfinger JR, Gullans SR: Global analysis of gene expression in renal ischemia-reperfusion in the mouse. Biochem Biophys Res Commun 2002;291:787–794.
  18. Shiraki-Iida T, Iida A, Nabeshima Y, Anazawa H, Nishikawa S, Noda M, Kuro-o M, Nabeshima Y: Improvement of multiple pathophysiological phenotypes of klotho (kl/kl) mice by adenovirus-mediated expression of the klotho gene. J Gene Med 2000;2:233–244.
  19. Saito Y, Nakamura T, Ohyama Y, Suzuki T, Iida A, Shiraki-Iida T, Kuro-o M, Nabeshima Y, Kurabayashi M, Nagai R: In vivo klotho gene delivery protects against endothelial dysfunction in multiple risk factor syndrome. Biochem Biophys Res Commun 2000;267:767–772.
  20. Mizuno I, Takahashi Y, Okimura Y, Kaji H, Chihara K: Upregulation of the klotho gene expression by thyroid hormone and during adipose differentiation in 3T3-L1 adipocytes. Life Sci 2001;68:2917–2923.
  21. Kato Y, Arakawa E, Kinoshita S, Shirai A, Furuya A, Yamano K, Nakamura K, Iida A, Anazawa H, Koh N, Iwano A, Imura A, Fujimori T, Kuro-o M, Hanai N, Takeshige K, Nabeshima Y: Establishment of the anti-klotho monoclonal antibodies and detection of klotho protein in kidneys. Biochem Biophys Res Commun 2001;267:597–602.
  22. Saito K, Ishizaka N, Mitani H, Ohno M, Nagai R: Iron chelation and a free radical scavenger suppress angiotensin II-induced downregulation of klotho, an anti-aging gene, in rat. FEBS Lett 2003;551:58–62.
  23. Nagai T, Yamada K, Kim HC, Kim YS, Noda Y, Imura A, Nabeshima Y: Cognition impairment in the genetic model of aging klotho gene mutant mice: a role of oxidative stress. FASEB J 2002;17:50–52.

    External Resources

  24. Mitani H, Ishizaka N, Aizawa T, Ohno M, Usui S, Suzuki T, Amaki T, Mori I, Nakamura Y, Sato M, Nangaku M, Hirata Y, Nagai R: In vivo klotho gene transfer ameliorates angiotensin II-induced renal damage. Hypertension 2002;39:838–843.
  25. Nakamura T, Saito Y, Ohyama Y, Masuda H, Sumino H, Kuro-o M, Nabeshima Y, Nagai R, Kurabayashi M: Production of nitric oxide, but not prostacyclin, is reduced in klotho mice. Jpn J Pharmacol 2002;89:149–156.

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