Quantitation of intracellular oxidation in a renal epithelial cell line

We quantitated the presence of intracellular oxidizing species in response to oxidative stimuli using fluorescent cell analytic techniques

J Scott

2003

Scholarcy highlights

  • We quantitated the presence of intracellular oxidizing species in response to oxidative stimuli using fluorescent cell analytic techniques
  • Oxygen radical formation was initiated by the addition of FeCl2 or xanthine oxidase to the culture media
  • Increases in intracellular dichlorofluorescin diacetate oxidation in response to xanthine oxidase exposure were inhibited by extracellular superoxide dismutase, catalase and dimethyl sulfoxide, implicating the superoxide anion, hydrogen peroxide, and the hydroxyl radical in producing the changes in intracellular dichlorofluorescent in fluorescence
  • Increases in intracellular DCFDA oxidation in response to xanthine oxidase correlated with loss of cellular viability as established by decreased plating efficiency
  • We conclude that relative intracellular oxidation can be quantitated within the cultured renal cell and that some extracellularly generated radicals may be capable of traversing the intact cell membrane to oxidize DCFDA in the cell interior
  • We conclude that relative intracellular oxidation can be quantitated within the cultured renal cell and that some extracellularly generated radicals may be capable of traversing the intact cell membrane to oxidize dichlorofluorescin diacetate in the cell interior

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