Critical role of mitochondria, but not caspases, during glucocorticosteroid-induced human eosinophil apoptosis.

Glucocorticosteroids are potent anti-inflammatory drugs used in the treatment of eosinophilic disorders. These molecules directly promote eosinophil apoptosis, yet the molecular mechanisms regulating this process remain ill-defined. We show here that stimulation of human peripheral blood eosinophils with dexamethasone induced DNA fragmentation, chromatin and cytoplasm condensation, and caspase-3 activation, as assessed by the proteolysis of its zymogen form and by the increase of caspase-3-like activity in eosinophil lysates. These phenomena were accompanied by a reduced uptake of the mitochondrial potential-sensitive marker DiOC(6)(3), suggestive of mitochondrial membrane permeabilization. Eosinophil incubation with the caspase-3 inhibitor, Z-Asp-Glu-Val-Asp-fluromethylketone, or with the broad spectrum caspase inhibitor, Z-Val-Ala-Asp-fluromethylketone, inhibited caspase-3-like activity generation but failed to modify dexamethasone-mediated loss in mitochondrial transmembrane potential and eosinophil apoptosis. In contrast, bongkrekic acid, a ligand of the mitochondrial permeability transition pore component, adenine nucleotide translocator, prevented both dexamethasone-induced mitochondrial disruption and apoptosis. We conclude that the mitochondrial permeability transition pore, rather than the caspase cascade, plays a critical role in the propagation of glucocorticosteroid-mediated apoptotic signals in human eosinophils.