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

Abstract

1 Non-steroidal anti-in¯ammatory 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 cyclooxygenase-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 mM for 24 h) promoted cell loss, and apoptosis, under both basal conditions and when apoptosis was increased by 25 mM N-Hexanoyl-D-sphingosine (C 6 -ceramide). 4 Diclofenac (250 mM 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 mM C 6 -ceramide. 5 The inhibitory eect of diclofenac on basal caspase-3-like activity was also exhibited by the structurally similar mefenamic and¯ufenamic acids (1 ± 250 mM), but not by ni¯umic 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 aect 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 eect on apoptotic activity in a primary culture of guineapig gastric mucous epithelial cells, and the inhibitory eect of diclofenac on basal apoptosis could involve an action on caspase activity.

Enhanced Death Ligand-Induced Apoptosis in Cutaneous SCC Cells by Treatment with Diclofenac/Hyaluronic Acid Correlates with Downregulation of c-FLIP

“…The role of diclofenac in apoptosis regulation has, however, been controversial. Even antiapoptotic effects have been attributed as suppression of ER stress-induced apoptosis in neuroblastoma cells (Yamazaki et al, 2006) and inhibition of basal apoptosis in guinea-pig gastric mucous cells (Ashton and Hanson, 2002).…”

Potential Protective Effects of NSAIDs/ASA in Oxidatively Stressed Human Lens Epithelial Cells and Intact Mouse Lenses in Culture

“…It has been shown that in rat liver cells, celecoxib and indomethacin stimulate the release of arachidonic acid, which may be associated with induction of apoptosis [27] . It has also been demonstrated that indomethacin and to some extent diclofenac cause apoptosis in chick embryo fi broblasts in a dose-and time-dependent manner [28] , whereas Ashton and Hanson [29] showed that diclofenac reduced the portion of apoptotic nuclei in guinea-pig gastric mucous cells. These studies in other tissues as well as the present study, indicate that the concentration is crucial.…”

Intracellular Effects of NSAIDs/ASA in Oxidatively Stressed Human Lens Epithelial Cells in Culture

“…Studies in the eye as well as in other tissues have found protective as well as damaging effects by NSAIDs/ASA [11,23,[31][32][33][34][35][36] . In a previous study, we found protection against hydrogen peroxide-induced apoptosis by NSAIDs/ASA at low concentrations, whereas higher concentrations were toxic, emphasizing the importance of the chosen concentration [23] .…”