Dissociated ROS production and ceramide generation in sulfasalazine-induced cell death in Raw 264.7 cells.
Sulfasalazine (SSZ) is a drug used in inflammatory bowel disease, whose precise mechanism of action remains to be clarified. Here, we report that incubation of Raw 264.7 cells with SSZ but not salicylates [acetylsalicylic acid (ASA), 4-aminosalicylic acid (4-ASA), and 5-ASA] causes a mixed apoptotic and necrotic form of cell death. In contrast to its metabolites, sulfapyridine and 5-ASA, SSZ exposure in Raw 264.7 cells resulted in a threefold increase in ceramide generation, as well as a robust production of reactive oxygen species (ROS). However, inhibition of ceramide production by fumonisin B1 failed to attenuate cell death. Preincubation with catalase, cyclosporin A (CsA), and bongkrekic acid attenuated ROS production. When dead cells were quantified for apoptotic versus necrotic cell death, catalase and N-acetylcysteine reproducibly attenuated apoptosis, whereas CsA, in addition to reducing apoptosis, was observed to dramatically enhance necrosis. In conclusion, the cell-death response induced by SSZ in Raw 264.7 cells involves ROS in the apoptotic limb but is independent of ceramide formation.
Salh
B
B
The Jack Bell Research Centre, Vancouver, British Columbia, Canada. bsalh@interchange.ubc.ca
Assi
K
K
Huang
S
S
O'Brien
L
L
Steinbrecher
U
U
Gómez-Muñoz
A
A
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D017209:Apoptosis; D002460:Cell Line; D002518:Ceramides / Q000096:biosynthesis*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D051379:Mice; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction; D012460:Sulfasalazine / Q000494:pharmacology*
D000894:Anti-Inflammatory Agents, Non-Steroidal; D002518:Ceramides; D017382:Reactive Oxygen Species; D012460:Sulfasalazine