Hydrogen peroxide induced oxidative stress damage and antioxidant enzyme response in Caco-2 human colon cells

Subhashinee S.K. Wijeratne, Susan L. Cuppett, Vicky Schlegel

Research output: Contribution to journalArticle

137 Scopus citations

Abstract

Studies were conducted to evaluate the cell damage caused by exposing human colon carcinoma cells, Caco-2, to hydrogen peroxide at concentrations varying from 0 to 250 μM for 30 min. Evaluation of cell viability, as measured by trypan blue dye exclusion test, showed that the loss of viability was <5% at concentrations up to 250 μM hydrogen peroxide. Cell membrane damage and DNA damage as measured by the leakage of lactate dehydrogenase and the comet assay, respectively, were significantly high at concentrations >100 μM hydrogen peroxide compared to those of the control. Antioxidant mechanisms in Caco-2 cells were evaluated by measuring catalase, superoxide dismutase, and glutathione peroxidase activities. Catalase activities remained constant in cells treated with 50-250 μM hydrogen peroxide. Superoxide dismutase activity decreased, whereas glutathione peroxidase activity increased in cells treated with H2O2 concentrations of >50 μM. This study showed that with increasing hydrogen peroxide concentration, cell membrane leakage and DNA damage increased, whereas the three antioxidant enzymes responded differently, as shown by mathematical models.

Original languageEnglish (US)
Pages (from-to)8768-8774
Number of pages7
JournalJournal of Agricultural and Food Chemistry
Volume53
Issue number22
DOIs
StatePublished - Nov 2 2005

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Keywords

  • Antioxidant enzymes
  • Caco-2
  • Catalase
  • Cell viability
  • Colon
  • DNA damage
  • Glutathione peroxidase
  • Hydrogen peroxide
  • Oxidative stress
  • Superoxide dismutase

ASJC Scopus subject areas

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

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