Tert-butyl hydroperoxide damages mitochrondria and induces apoptosis in cortical neurons

AIM: To explore the possible mechanism of tert - butyl hydroperoxide (t - BHP) - induced apoptosis in rat cortical neurons. METHODS: Primary cultured rat cortical neurons were performed in vitro and cell viability was measured by MTT assay. DNA fragmentation was used to evaluate cell apoptosis and mitochondrial transmembrane potential △(?)m) was determined by flow cytometric assay. Cellular glutathione (GSH) content was measured by spectrophotometer. Bcl- 2 and Bax protein, cytosolic cytochrome c, cleaved caspase- 3 and poly (ADP- ribose) polymerase (PARP) were detected by Western blotting. RESULTS: After exposure of cortical neurons to tBHP (25 - 400 μmol/L), the cell viability was reduced. △(?)m and cellular GSH content were also decreased significantly. The level of Bcl - 2 protein was reduced and Bax was elevated. Meanwhile, tBHP exposure resulted in cytochrome c release, caspase - 3 and PARP proteolysis, DNA fragmentation and eventually neuron apoptosis. CONCLUSION: Mitochondrial damage may mediate tBHP - induced apoptosis in cortical neurons.