Some oxysterols are associated with neurodegenerative diseases. Their lipotoxicity is characterized by an oxidative stress and induction of apoptosis. To evaluate the capacity of these molecules to trigger cellular modifications involved in neurodegeneration, human neuronal cells SK-N-BE were treated with 7-ketocholesterol, 7 alpha- and 7 beta-hydroxycholesterol, 6 alpha- and 6 beta-hydroxycholesterol, 4 alpha- and 4 beta-hydroxycholesterol, 24(S)-hydroxycholesterol and 27-hydroxycholesterol (50-100 mu M, 24 h) without or with docosahexaenoic acid (50 mu M). The effects of these compounds on mitochondrial activity, cell growth, production of reactive oxygen species (ROS) and superoxide anions (O-2(-)), catalase and superoxide dismutase activities were determined. The ability of the oxysterols to induce increases in Ca2+ was measured after 10 min and 24 h of treatment using fura-2 videomicroscopy and Von Kossa staining, respectively. Cholesterol, 7-ketocholesterol, 7 beta-hydroxycholesterol, and 24(S)-hydroxycholesterol (100 mu M) induced mitochondrial dysfunction, cell growth inhibition, ROS overproduction and cell death. A slight increase in the percentage of cells with condensed and/or fragmented nuclei, characteristic of apoptotic cells, was detected. With 27-hydroxycholesterol, a marked increase of O-2(center dot-) was observed. Increases in intracellular Ca2+ were only found with 7-ketocholesterol, 7 beta-hydroxycholesterol, 24(S)-hydroxycholesterol and 27-hydroxycholesterol. Pre-treatment with docosahexaenoic acid showed some protective effects depending on the oxysterol considered. According to the present data, 7-ketocholesterol, 7 beta-hydroxycholesterol, 24(S)-hydroxycholesterol and 27-hydroxycholesterol could favor neurodegeneration by their abilities to induce mitochondrial dysfunctions, oxidative stress and/or cell death associated or not with increases in cytosolic calcium levels.