Reactive oxygen species induce apoptosis in cultured human mesangial cells

Hitoshi Sugiyama, Naoki Kashihara, Hirofumi Makino, Yasushi Yamasaki, Zensuke Ota

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Abstract

Apoptosis is a distinct form of cell death that is observed under various physiologic and pathologic conditions, and it is thought to be important in regulating the number of glomerular cells. This study investigated the possible role of reactive oxygen species in the induction of apoptosis in cultured human mesangial cells. Fragmented nuclei with condensed chromatin, a morphologic characteristic of apoptosis, were observed by electron microscopy in mesangial cells exposed to 0.02 mM hydrogen peroxide for 4 h. Nuclear DNA extracted from mesangial cells that had been incubated with hydrogen peroxide (2 to 20 mM) or with xanthine (0.05 mM) and xanthine oxidase (5 to 100 mU/mL) showed the ladder pattern on electrophoresis that is a biochemical marker for apoptosis. Hydrogen peroxide (0.02 to 20 mM) decreased the number of viable cells, as determined by trypan blue exclusion, in a dose-dependent manner. Hydrogen peroxide or xanthine and xanthine oxidase increased the lactate dehydrogenase release from mesangial cells in a dose- and time-dependent manners. The release of lactate dehydrogenase was prevented by treatment with a free radical scavenger, catalase. Hydrogen peroxide (2 mM) also significantly increased the number of mesangial cells with fragmented DNA as detected by in situ nick end-labeling. Results indicate that reactive oxygen species induce apoptosis in cultured human mesangial cells. Furthermore, apoptosis of mesangial cells induced by reactive oxygen species may contribute to the loss of such cells observed in glomerular disease.

Original languageEnglish
Pages (from-to)2357-2363
Number of pages7
JournalJournal of the American Society of Nephrology
Volume7
Issue number11
Publication statusPublished - Nov 1996

Fingerprint

Mesangial Cells
Reactive Oxygen Species
Apoptosis
Hydrogen Peroxide
Xanthine
Xanthine Oxidase
L-Lactate Dehydrogenase
Cell Count
Free Radical Scavengers
Trypan Blue
DNA
In Situ Nick-End Labeling
Catalase
Chromatin
Electrophoresis
Electron Microscopy
Cell Death
Biomarkers

Keywords

  • Cell death
  • DNA fragmentation
  • Hydrogen peroxide
  • Xanthine
  • Xanthine oxidase

ASJC Scopus subject areas

  • Nephrology

Cite this

Reactive oxygen species induce apoptosis in cultured human mesangial cells. / Sugiyama, Hitoshi; Kashihara, Naoki; Makino, Hirofumi; Yamasaki, Yasushi; Ota, Zensuke.

In: Journal of the American Society of Nephrology, Vol. 7, No. 11, 11.1996, p. 2357-2363.

Research output: Contribution to journalArticle

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