Disulfiram inhibits TNF-alpha-induced cell death

Disulfiram, a clinically employed alcohol deterrent, was recently discovere d to inhibit caspase-3 and DNA fragmentation. Using LLC-PK1 cells and murin e liver as models, we examined if the drug inhibited TNF-alpha-induced cell death. Disulfiram produced dose-dependent inhibition of TNF-alpha-induced cell death as well as caspase-3-like activity, Disulfiram retained 80% of i ts effect when added 4 h after TNF-alpha. Disulfiram protected the cells fr om cytokine-induced death for at least 6 days. The cells rescued by the dru g preserved the ability to proliferate. The cells died spontaneously after exposure to TNF-alpha for just 70 min. Co-administration of 15 mu M disulfi ram and TNF-alpha for 70 min prior to their removal abolished TNF-alpha-ind uced killing, and this was associated with restoration of mitochondrial mem brane potential and suppression of reactive oxygen species. Treatment of mi ce with TNF-alpha and D-galactosamine for 5 h markedly increased hepatic DN A fragmentation and caspase-3-like activity. Disulfiram at 0.6 mmol/kg abol ished these effects. We conclude that disulfiram is a potent inhibitor of T NF-alpha-induced cell death in vitro. The underlying mechanisms include sta bilization of mitochondrial membrane potential, suppression of reactive oxy gen species, and inhibition of caspase-3-like activity. We further conclude that disulfiram inhibits DNA fragmentation in vivo in association with the blockade of caspase-3-like activity.