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Production of Hydrogen Peroxide During Hypoxia-Reoxygenation in Isolated Myocytes

  • Ryuji Ueda
  • Noburu Konno
  • Masaki Nakatani
  • Toshiki Iwata
  • Yoshihisa Arata
  • Katsumiti Iijima
  • Kazuaki Nishio
  • Eiichi Geshi
  • Toshikuni Yanagishita
  • Takashi Katagiri
  • Yasumitsu Nakai
Part of the Progress in Experimental Cardiology book series (PREC, volume 1)

Abstract

Active oxygen species, including hydrogen peroxide (H2O2), have been implicated in myocardial reperfusion injury. Recently, spin-trap agents and biochemical techniques applied to intact hearts have shown that H2O2 is generated by leukocytes, by endothelial cells, and by mitochondria in myocytes. In this study, we used electron microscopy and the cerium (Ce) method to histologically investigate H2O2 formation during hypoxia—reoxygenation and its toxic effects on myocardium. This Ce method involves the formation of an electron-dense precipitate when H2O2 reacts with cerium chloride (CeCl3). Single myocytes were obtained from rat hearts by the collagenase method. Isolated myocytes were reoxygenated for 15 minutes after 30 minutes of hypoxia. Digitonin and CeCl3, were added to make cell membranes permeable and to detect intracellular H2O2 by electron microscopy. In the control group, the ultrastructure was well preserved and no dense deposits were found in myocytes. However, in the hypoxia—reoxygenation group, precipitates, which were cerium—H2O2 reaction products, were found along swollen mitochondria. Moreover, in the hypoxia-reoxygenation group, cell viability was reduced to 72% of control. These results indicate that H2O2 is generated by mitochondria and that its relese into cytosol may lead to myocyte death during hypoxia—reperfusion.

Keywords

Xanthine Oxidase Reactive Oxygen Intermediate Dense Deposit Swell Mitochondrion Light Microscopic Finding 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Ryuji Ueda
    • 1
  • Noburu Konno
    • 1
  • Masaki Nakatani
    • 1
  • Toshiki Iwata
    • 1
  • Yoshihisa Arata
    • 1
  • Katsumiti Iijima
    • 1
  • Kazuaki Nishio
    • 1
  • Eiichi Geshi
    • 1
  • Toshikuni Yanagishita
    • 1
  • Takashi Katagiri
    • 1
  • Yasumitsu Nakai
    • 1
  1. 1.Showa University School of MedicineJapan

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