Oxidative insults and HNE induce apoptosis in primary hippocampal neurons: prevention by GSH. A, Cultures were exposed to 2 μm of each aldehyde (see Fig. 2C for aldehyde structures) or 0.2% ethanol (Control) for 16 hr, and percentages of neurons with condensed and fragmented nuclei were quantified. Values are the mean and SEM of determinations made in four separate cultures. *p < 0.05, **p < 0.001 compared with control value (ANOVA with Scheffé’s post hoc tests). B, Cultures were exposed for 16 hr to 0.2% ethanol (Control) or the indicated treatments, at which time cells were stained with Hoescht dye and the percentages of neurons with condensed and fragmented nuclei were quantified. Treatment concentrations were HNE, 10 μm; Aβ, 10 μm;FeSO4, 2 μm;GSH, 1 mm. Values are the mean and SEM of determinations made in four separate cultures. *p< 0.01 compared with control value; **p < 0.01, ***p < 0.05 compared with corresponding values for cultures not cotreated with GSH (ANOVA with Scheffé’spost hoc tests). C, Images of Hoescht dye fluorescence in hippocampal neurons from cultures exposed for 16 hr to 0.2% ethanol (Control), 2 μmHNE, 1 mm GSH-ethyl-ester plus 2 μm HNE (GSH+HNE), or 10 μmAβ. HNE and Aβ induced nuclear condensation and fragmentation in most neurons (arrowheads), whereas fluorescence remained uniformly distributed throughout the nucleus in neurons in control cultures and neurons in cultures cotreated with GSH and HNE (arrowheads).
