PMC

Binge ethanol-dependent PI labeling of degenerating neurons in the EC and DG in representative organotypic HEC slices in culture and suppression by MEP or by DHA. (A) Control slice; (B) slice supplemented with DHA (25 µM); (C) slice supplemented with MEP (1 µM); (D) binge ethanol (100 mM)-treated slice; (E) binge ethanol-treated and MEP-supplemented slice; (F) binge ethanol-treated and DHA-supplemented slice

Mobilization of [³H] from incorporated [³H]AA in HEC slices in culture is significantly increased by ethanol + withdrawal treatment and normalized by supplementation with DHA. Results expressed as cpm/mg slice protein. N = 6–9 wells/grp. See legend for explanation of box plot. *Groups whose means were significantly less (P < .05) than the mean of the ethanol (E) group by Holm-Bonferroni t-tests. Overall one-way ANOVA was significant (F_3,18 = 7.91, P = .0014)

Neurodegeneration in the EC is significantly increased compared to control (Cont) by binge ethanol (E) treatment and inhibited by supplementation with DHA but not ADA. Neurodegeneration expressed as percent PI labeling. N = 6–9 wells/grp. See legend for explanation of box plots. *Groups whose means were significantly less (P <. 05) than the mean of the ethanol (E) group by Holm-Bonferroni t-test. Overall one-way ANOVA was significant (F_5,36 = 6.77, P = .00015)

Proposed mechanistic scheme for binge ethanol-induced neurodegeneration. Upper pathway depicts focus of this report: brain edema, PLA2 activation, AA mobilization, and ROS generation, and inhibition (neuroprotection) by diuretics, MEP, DHA, and ROS trapping agents at various potential steps. The roles of edema and ROS in the model are predicated on the basis of in vivo results with binge ethanol rat models [, , ]. Two lower pathways depict possible ROS generation during ethanol exposure from increased NADPH oxidase activity and mitochondrial leakage/damage. See text for further discussion

Neurodegeneration in HEC slices in culture is significantly increased over control (Cont) by binge ethanol (E) treatment and inhibited by supplementation with MEP. Box in boxplot illustrates the extent of the middle two quartiles and defines the interquartile range (IQR), along with the median (horizontal line). Filled circle inside box shows the mean with error bars showing SEM. Whiskers extending from box depict maximum range of values within 1.5 IQR, while small open circles denote “outlier” values. N = 6–9 wells/grp. (A) Neurodegeneration expressed as percent PI labeling. *Group mean was significantly greater (P < .01) than the mean of the Control (Cont) group by Holm-Bonferroni t-tests. Overall one-way ANOVA was significant (F_3.20 = 65.34, P = 2.03 × 10⁻¹⁴). (B) Neurodegeneration expressed as media LDH activity/mg slice protein. *Group mean significantly greater (P < .05) than the mean of the Control (Cont) group by Holm-Bonferroni t-test. ^@Group mean significantly less (P <0.05) than the mean for the Ethanol (E) group by Holm-Bonferroni t-test. Overall one-way ANOVA was significant (F_3,25 = 21.04, P = 2.13 × 10⁻¹⁶)