Disparate effects of non-steroidal anti-inflammatory drugs on apoptosis in guinea-pig gastric mucous cells: inhibition of basal apoptosis by diclofenac

1 Non-steroidal anti-inflammatory drugs (NSAIDs) induce apoptosis in gastrointestinal cancer cell lines. Similar actions on normal gastric epithelial cells could contribute to NSAID gastropathy. The present work therefore compared the actions of diclofenac, ibuprofen, indomethacin, and the cyclo-oxygenase-2 selective inhibitor, NS-398, on a primary culture of guinea-pig gastric mucous epithelial cells. 2 Cell number was assessed by staining with crystal violet. Apoptotic activity was determined by condensation and fragmentation of nuclei and by assay of caspase-3-like activity. Necrosis was evaluated from release of cellular enzymes. 3 Ibuprofen (250 mum for 24 h) promoted cell loss, and apoptosis, under both basal conditions and when apoptosis was increased by 25 muM N-Hexanoyl-D-sphingosine (C-6-ceramide). 4 Diclofenac (250 muM for 24 h) reduced the proportion of apoptotic, nuclei from 5.2 to 2.1%, and caused inhibition of caspase-3-like activity, without causing necrosis under basal conditions. No such reduction in apoptotic activity was evident in the presence of 25 muM C-6-ceramide. 5 The inhibitory effect of diclofenac on basal caspase-3-like activity was also exhibited by the structurally similar mefenamic and flufenamic acids (1 -250 muM), but not by niflumic acid. 6 Inhibition of superoxide production by the cells increased caspase-3-like activity, but the inhibitory action of diclofenac on caspase activity remained. Diclofenac did not affect superoxide production. 7 Diclofenac inhibited caspase-3-like activity in cell homogenates and also inhibited human recombinant caspase-3. 8 In conclusion, NSAIDs vary in their effect on apoptotic activity in a primary culture of guinea pig gastric mucous epithelial cells, and the inhibitory effect of diclofenac on basal apoptosis could involve an action on caspase activity.