Glutathione oxidation and PTPase inhibition by hydrogen peroxide in Caco-2 cell monolayer

Radhakrishna Rao, L. Li, R. D. Baker, S. S. Baker, A. Gupta

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

The role of H2O2 and protein thiol oxidation in oxidative stress-induced epithelial paracellular permeability was investigated in Caco-2 cell monolayers. Treatment with a H2O2 generating system (xanthine oxidase + xanthine) or H2O2 (20 μM) increased the paracellular permeability. Xanthine oxidase-induced permeability was potentiated by superoxide dismutase and prevented by catalase. H2O2-induced permeability was prevented by ferrous sulfate and potentiated by deferoxamine and 1,10-phenanthroline. GSH, N-acetyl-L-cysteine, dithiothreitol, mercaptosuccinate, and diethylmaleate inhibited H2O2-induced permeability, but it was potentiated by 1,3-bis(2-chloroethyl)-1-nitrosourea. H2O2 reduced cellular GSH and protein thiols and increased GSSG. H2O2-mediated reduction of GSH-to-GSSG ratio was prevented by ferrous sulfate, GSH, N-acetyl-L-cysteine, diethylmaleate, and mercaptosuccinate and potentiated by 1,10-phenanthroline and 1,3-bis(2-chloroethyl)-1-nitrosourea. Incubation of soluble fraction of cells with GSSG reduced protein tyrosine phosphatase (PTPase) activity, which was prevented by co-incubation with GSH. PTPase activity was also lower in H2O2-treated cells. This study indicates that H2O2, but not O2 - · or · OH, increases paracellular permeability of Caco-2 cell monolayer by a mechanism that involves oxidation of GSH and inhibition of PTPases.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume279
Issue number2 42-2
StatePublished - Sep 14 2000
Externally publishedYes

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Protein Tyrosine Phosphatases
Caco-2 Cells
Hydrogen Peroxide
Glutathione
Permeability
ferrous sulfate
diethyl maleate
Glutathione Disulfide
Carmustine
Xanthine Oxidase
Acetylcysteine
Sulfhydryl Compounds
Xanthine
Deferoxamine
Dithiothreitol
Catalase
Superoxide Dismutase
Proteins
Oxidative Stress

All Science Journal Classification (ASJC) codes

  • Physiology
  • Hepatology
  • Gastroenterology
  • Physiology (medical)

Cite this

Glutathione oxidation and PTPase inhibition by hydrogen peroxide in Caco-2 cell monolayer. / Rao, Radhakrishna; Li, L.; Baker, R. D.; Baker, S. S.; Gupta, A.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 279, No. 2 42-2, 14.09.2000.

Research output: Contribution to journalArticle

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