Ceramide induces a loss in cytosolic peroxide levels in mononuclear cells

0038, Article English OPEN
Phillips, Darren C. ; Griffiths, Helen R. (2003)

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.
  • References (44)
    44 references, page 1 of 5

    1. Gamard, C., Dbaibo, G., Liu, B., Obeid, L. and Hannun, Y. (1997). Selective involvement of ceramide in cytokine-induced apoptosis. J . Biol. Chem. 272, 16474-16481.

    2. Liu, B., Andreieu-Abadie, N., Levade, T., Zhang, P., Obeid, L. and Hannun, Y. (1998). Glutathione regulation of neutral sphingomyelinase in Tumour necrosis factor-a induced cell death. J. Biol. Chem. 273, 11313-11320.

    3. Obeid, L., Linardic, C., Karolak, L. and Hannun, Y. (1993). Programmed cell death induced by ceramide. Science 259, 1769-1771.

    4. Verheij, M., Bose, R., Lin, X., Yao, B., Jarvis, W., Grant, S., Birrer, M., Szabo, E., Zon, L., Kyriakis, J., Haimovitz-Friedman, A., Fuks, Z. and Kolesnick, R. (1996). Requirement for ceramide initiated SAPK/JNK signaling in stress-induced apoptosis. Nature 380, 75-79.

    5. Andrieu, N., Salvayre, R. and Levade, T. (1994). Evidence against involvement of the acid lysosomal sphingomyelinase in the tumor-necrosis-factor- and interleukin-1-induced sphingomyelin cycle and cell proliferation in human fibroblasts. Biochem J. 303, 341-345.

    6. Cifone, M., De Maria, R., Roncaioli, P., Rippo, M., Azuma, M., Lanier, L., Santoni, A. and Testi, R. (1993). Apoptotic signalling through CD95 (Fas/Apo-1) activates an acidic sphingomyelinase. J. Exp. Med. 177, 1547-1552.

    7. Gulbins, E., Bissonnette, R., Mahboubi, A., Martin, S., Nishioka, W., Brunner, T., Baier, G., Baier-Bitterlich, G., Byrd, C., Lang, F., Kolesnick, R., Altman, A. and Green, D. (1995). FAS-induced apoptosis is mediated via a ceramide-initiated RAS signalling pathway. Immunity 2, 341-351.

    8. Tepper, A., Boesen-De Cock, J.G.R., De Vries, E., Borst, J. And Van Blitterswijk, W.J. (1997). CD95/Fas-induced ceramide formation proceeds with slow kinetics and is not blocked by caspase-3/CPP32 inhibition. J. Biol. Chem. 272, 24308-24312.

    9. Santana, P., Peña, L., Haimovitz-Friedman, A., Martin, S., Green, D., Mcloughlin, M., Cordon-Cardo, C., Schuchman, E., Fuks, Z. and Kolesnick, R. (1996). Acid sphingomyelinase-deficient human lymphoblasts and mice are defective in radiation-induced apoptosis. Cell 86, 189-199.

    10. Boland, M., Foster, S. and O'neill, L. (1997). Daunorubicin activates NFκB and induces κB-dependent gene expression in HL-60 promyelocytic and Jurkat T lymphoma cells. J. Biol. Chem. 272, 12952-12960.

  • Metrics
    No metrics available
Share - Bookmark