FIGURE 9.

Cardanol pretreatment sensitizes primary cortical neurons to oxidative insult. Rat embryonic day 18 cortical neurons were pretreated for 1–2 h with cardanol prior to an overnight (16–18-h) tBHP exposure. The cardanol stock solution was prediluted 1:5 in 2-hydroxypropyl-β-cyclodextrin prior to dilution in medium to enhance delivery. Neuronal viability was measured by calcein fluorescence and normalized to the vehicle control. A, cardanol pretreatment exacerbates tBHP-mediated cell death. B, the bioactive lipid NAE 16:0 (palmitoylethanolamide, N-(2-hydroxyethyl)hexadecanamide), a substrate of FAAH, reverses this effect. Cardanol treatment significantly reduces neuronal viability in antioxidant-free medium and exacerbates tBHP-mediated cell death. C and D, preincubation with the specific, irreversible FAAH inhibitors MAFP and URB597 block this effect of cardanol. Data points represent means ± S.E. (error bars) of triplicate assays. Plots were generated with GraphPad Prism version 5.0. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001, as determined by one-way analysis of variance with Dunnett's post-test.