Ceramide induces a loss in cytosolic peroxide levels in mononuclear cells

Darren C. Phillips, Helen R. Griffiths*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Ceramide (a sphingolipid) and reactive oxygen species are each partly responsible for intracellular signal transduction in response to a variety of agents. It has been reported that ceramide and reactive oxygen species are intimately linked and show reciprocal regulation [Liu, Andreieu-Abadie, Levade, Zhang, Obeid and Hannun (1998) J. Biol. Chem. 273, 11313-11320]. Utilizing synthetic, short-chain ceramide to mimic the cellular responses to fluctuations in natural endogenous ceramide formation or using stimulation of CD95 to induce ceramide formation, we found that the principal redox-altering property of ceramide is to lower the [peroxide]cyt (cytosolic peroxide concentration). Apoptosis of Jurkat T-cells, primary resting and phytohaemagglutinin-activated human peripheral blood T-lymphocytes was preceded by a loss in [peroxide]cyt, as measured by the peroxide-sensitive probe 2′,7′-dichlorofluorescein diacetate (also reflected in a lower rate of superoxide dismutase-inhibitable cytochrome c reduction), and this was not associated with a loss of membrane integrity. Where growth arrest of U937 monocytes was observed without a loss of membrane integrity, the decrease in [peroxide]cyt was of a lower magnitude when compared with that preceding the onset of apoptosis in T-cells. Furthermore, decreasing the cytosolic peroxide level in U937 monocytes before the application of synthetic ceramide by pretreatment with either of the antioxidants N-acetyl cysteine or glutathione conferred apoptosis. However, N-acetyl cysteine or glutathione did not affect the kinetics or magnitude of ceramide-induced apoptosis of Jurkat T-cells. Therefore the primary redox effect of cellular ceramide accumulation is to lower the [peroxide]cyt of both primary and immortalized cells, the magnitude of which dictates the cellular response.

Original languageEnglish
Pages (from-to)567-579
Number of pages13
JournalBiochemical Journal
Volume375
Issue numberPart 3
DOIs
Publication statusPublished - 23 Jul 2003

Fingerprint

Ceramides
Peroxides
T-cells
Acetylcysteine
Apoptosis
T-Lymphocytes
Jurkat Cells
Oxidation-Reduction
Cysteine
Monocytes
Reactive Oxygen Species
Membranes
Signal transduction
Sphingolipids
Phytohemagglutinins
Cytochromes c
Superoxide Dismutase
Signal Transduction
Blood
Antioxidants

Bibliographical note

The final version of record is available at http://www.biochemj.org/bj/375/bj3750567.htm

Keywords

  • apoptosis
  • ceramide
  • cytosolic peroxide
  • growth arrest
  • reactive oxygen species
  • redox compartmentalization

Cite this

Phillips, Darren C. ; Griffiths, Helen R. / Ceramide induces a loss in cytosolic peroxide levels in mononuclear cells. In: Biochemical Journal. 2003 ; Vol. 375, No. Part 3. pp. 567-579.
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Ceramide induces a loss in cytosolic peroxide levels in mononuclear cells. / Phillips, Darren C.; Griffiths, Helen R.

In: Biochemical Journal, Vol. 375, No. Part 3, 23.07.2003, p. 567-579.

Research output: Contribution to journalArticle

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