The acid sphingomyelinase inhibitor SR33557 counteracts TNF-alpha-mediated potentiation of IL-1beta-induced NF-kappaB activation in the insulin-producing cell line Rinm5F.
Cytokines induce nitric oxide (NO) production and cell death in insulin-producing cells in vitro but the signaling pathways mediating the cytokine effects are not well characterized. The aim of this study was to determine whether sphingomyelinase (SMase) participates in cytokine signaling leading to NF-kappaB activation, iNOS induction and cell death in insulin-producing cells. Acute exposure to IL-1beta or TNF-alpha did not affect SMase activities in rat insulinoma (RINm5F) cells. TNF-alpha activated NF-kappaB in gel shift experiments without inducing iNOS--as assessed by nitrite formation--whereas IL-1beta stimulated both NF-kappaB activation and iNOS induction. Natural ceramide did not activate NF-kappaB or iNOS. However, both ceramide and TNF-alpha potentiated IL-1beta- induced activation of NF-kappaB and iNOS. Moreover, the potentiating effects of TNF-alpha were counteracted by the acid SMase inhibitor SR33557. The combination of IL-1beta and IFN-gamma induced apoptosis in RINm5F cells, which was paralleled by a modest increase in acid SMase, whereas ceramide mainly induced necrosis. It is concluded that cytokine-induced beta-cell signaling is associated with the induction of iNOS but not with enhanced SMase activities. However, TNF-alpha-mediated potentiation of the IL-1beta effect may involve an increased sensitivity to basal acid SMase activity. An increased acid SMase activity may participate in the execution of cytokine-induced beta-cell apoptosis.
Saldeen
J
J
Department of Medical Cell Biology, Uppsala University, Sweden.
Jaffrézou
J P
JP
Welsh
N
N
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002518:Ceramides; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D007212:Indolizines / Q000494:pharmacology*; D007328:Insulin / Q000096:biosynthesis; D007375:Interleukin-1 / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D009569:Nitric Oxide / Q000096:biosynthesis; D010627:Phenethylamines / Q000494:pharmacology*; D051381:Rats; D015398:Signal Transduction / Q000276:immunology*; D013108:Sphingomyelin Phosphodiesterase / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D002518:Ceramides; D004791:Enzyme Inhibitors; D007212:Indolizines; D007328:Insulin; D007375:Interleukin-1; D016328:NF-kappa B; D010627:Phenethylamines; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D013108:Sphingomyelin Phosphodiesterase; C059901:fantofarone