Thiol-mediated redox regulation of neutrophil apoptosis

R. W G Watson, O. D. Rotstein, A. B. Nathens, A. P B Dackiw, J. C. Marshall

Research output: Contribution to journalArticle

Abstract

Background. Intracellular glutathione, an endogenous antioxidant, protects cellular function against oxidative stress. Because oxidative stress has been implicated in neutrophil apoptosis, we hypothesized that reduced thiol levels may induce apoptosis through an alteration in cellular redox state. Methods. Human polymorphonuclear leukocytes (PMNs), were incubated with medium or with increasing concentrations of the reduced glutathione (GSH)-depleting agents diethylmaleate and diamide and buthionine sulfoximine, an inhibitor of GSH synthesis. Apoptosis was assessed by means of flow cytometry with propidium iodide DNA staining and confirmed morphologically. GSH was measured colorimetrically, and tyrosine phosphorylation was assessed by means of immunoblotting. Results. Diethylmaleate and diamide induced a dose-dependent reduction in GSH and a corresponding increase in PMN apoptosis. This effect could be reversed with N-acetylcysteine, suggesting that diethylmaleate induces apoptosis through the depletion of GSH. The antioxidant pyrolidine dithiocarbamate had no effect. Because oxidants can mediate intracellular signaling via tyrosine phosphorylation, we therefore evaluated the effects of the tyrosine kinase inhibition on diethylmaleate-induced PMN apoptosis. Both genistein and herbimycin A reduced diethylmaleate-induced apoptosis and tyrosine phosphorylation. Conclusions. Sulfhydryl oxidation by diethylmaleate alone induces apoptosis, providing evidence of a redox-sensitive, thiol-mediated pathway of apoptosis. Furthermore, tyrosine phosphorylation appears to play an important role in this process. Because apoptosis is a critical mechanism regulating PMN survival in vivo, manipulation of PMN intracellular thiols may represent a novel therapeutic target for the regulation of cellular function.

Original languageEnglish (US)
Pages (from-to)150-158
Number of pages9
JournalSurgery
Volume120
Issue number2
StatePublished - 1996
Externally publishedYes

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diethyl maleate
Sulfhydryl Compounds
Oxidation-Reduction
Neutrophils
Apoptosis
Tyrosine
Phosphorylation
Diamide
Glutathione
Oxidative Stress
Antioxidants
Buthionine Sulfoximine
Propidium
Genistein
Acetylcysteine
Immunoblotting
Oxidants
Protein-Tyrosine Kinases
Flow Cytometry

ASJC Scopus subject areas

  • Surgery

Cite this

Watson, R. W. G., Rotstein, O. D., Nathens, A. B., Dackiw, A. P. B., & Marshall, J. C. (1996). Thiol-mediated redox regulation of neutrophil apoptosis. Surgery, 120(2), 150-158.

Thiol-mediated redox regulation of neutrophil apoptosis. / Watson, R. W G; Rotstein, O. D.; Nathens, A. B.; Dackiw, A. P B; Marshall, J. C.

In: Surgery, Vol. 120, No. 2, 1996, p. 150-158.

Research output: Contribution to journalArticle

Watson, RWG, Rotstein, OD, Nathens, AB, Dackiw, APB & Marshall, JC 1996, 'Thiol-mediated redox regulation of neutrophil apoptosis', Surgery, vol. 120, no. 2, pp. 150-158.
Watson RWG, Rotstein OD, Nathens AB, Dackiw APB, Marshall JC. Thiol-mediated redox regulation of neutrophil apoptosis. Surgery. 1996;120(2):150-158.
Watson, R. W G ; Rotstein, O. D. ; Nathens, A. B. ; Dackiw, A. P B ; Marshall, J. C. / Thiol-mediated redox regulation of neutrophil apoptosis. In: Surgery. 1996 ; Vol. 120, No. 2. pp. 150-158.
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