Objective: Ceramide and sphingosine, generated by sphingomyelinase-mediated hydrolysis of sphingomyelin, which packs tightly in the bilayer of the plasma membrane, have been proposed as intracellular mediators of apoptotic signals. However, precise function of endogenous sphingomyelin-cycle metabolites in mast cells has been unclear. Thus, we sought to define the involvement of ceramide and sphingosine in apoptotic pathways of mast cells.
Materials and methods: We examined the effect of cell-permeable C(2)-ceramide, sphingosine, and sphingomyelinase on survival of murine bone marrow-derived cultured mast cells (BMCMC) supported by recombinant interleukin-3 (rIL-3) and/or recombinant stem cell factor (rSCF). Downstream signaling pathways of C(2)-ceramide and sphingosine were analyzed by using caspase inhibitors.
Results: C(2)-ceramide, sphingosine, and sphingomyelinase induced apoptosis in BMCMC in the presence of rIL-3 and/or rSCF, and Z-VAD-fmk (a broad caspase inhibitor), Z-DEVD-fmk (a caspase 3 inhibitor), and Z-IETD-fmk (a caspase 8 inhibitor) partially prevented apoptosis of BMCMC induced by C(2)-ceramide but not sphingosine.
Conclusion: The present results suggest that ceramide and sphingosine may function as intracellular mediators of apoptotic signals in mast cells, which override survival signals from IL-3 and SCF. In addition, caspases may be partially involved in ceramide- but not sphingosine-mediated apoptosis of mast cells.