tert-butylhydroperoxide induces peroxynitrite-dependent mitochondrial permeability transition leading PC12 cells to necrosis

A short-term exposure of PC12 cells to tert-butylhydroperoxide, followed by recovery in fresh culture medium, causes cell death and the extent of this response progressively increases during the 120 min of posttreatment incubation. Morphological and biochemical analyses of these cells revealed that the mode of cell death was necrosis. Cell killing induced by the hydroperoxide seems to be in part mediated by peroxynitrite because the lethal response was markedly and similarly reduced by the nitric oxide synthase inhibitor Nw-nitro-L-arginine methylester and by scavengers of nitric oxide or peroxynitrite. This peroxynitrite-dependent mechanism of cytotoxicity was blunted by antioxidants and inhibitors of mitochondrial permeability transition and the onset of cell death was preceded by mitochondrial depolarization and loss of cellular ATP. We conclude that tert-butylhydroperoxide promotes peroxynitrite-dependent PC12 cell necrosis causally linked to peroxidation of membrane lipids and mitochondrial permeability transition. (C) 2001 Wiley-Liss, Inc.