Prevention of 7-ketocholesterol-induced mitochondrial damage and cell death by calmodulin inhibition

Oxysterols such as 7-ketocholesterol and 25-hydroxycholesterol formed under enhanced oxidative stress in the brain are suggested to induce neuronal cell death. The present study investigated the effect of calmodulin antagonists (trifluoperazine, W-7 and calmidazolium) against the cytotoxicity of 7-ketocholesterol in relation to the mitochondria-mediated cell death process and oxidative stress. PC12 cells exposed to 7-ketocholesterol revealed nuclear damage, decrease in the mitochondrial transmembrane potential, cytosolic accumulation of cytochrome c, activation of caspase-3, increase in the formation of reactive oxygen species and depletion of GSH. N-Acetylcysteine, trolox, carboxy-PTIO and Mn-TBAP reduced the cytotoxic effect of 7-ketocholesterol. Calmodulin antagonists attenuated the 7-ketocholesterol-induced nuclear damage, formation of the mitochondrial permeability transition and cell viability loss in PC12 cells. The results suggest that calmodulin antagonists may prevent the 7-ketocholesterol-induced viability loss in PC12 cells by suppressing formation of the mitochondrial permeability transition, leading to the release of cytochrome c and subsequent activation of caspase-3. The effects seem to be ascribed to their depressant action on the formation of reactive oxygen species and depletion of GSH. The findings suggest that calmodulin inhibition may exhibit a protective effect against the neurotoxicity of 7-ketocholesterol.