Figure 3
From: Targeting cPLA2 derived lipid hydroperoxides as a potential intervention for sarcopenia
Loss of innervation primarily induces LOOH production through metabolism in the cytosolic phospholipase A2 (cPLA2) Pathway. (a) Dose–response of Amplex UltraRed fluorescence values in response to increasing concentrations of H2O2 or the LOOHs 15(S)-HpETE or 13(S)-HpODE. (b) Preliminary catalase inhibition dose–response curve in 7 day denervated muscle fibers (n = 2F). IC50 determined by variable slope (four parameters) test. (c) Basal hydroperoxide production rate in 7 day denervated fibers treated with catalase and/or GPX1 (n = 5–7 M). Statistical significance determined by ordinary one-way ANOVA with Tukey’s post hoc test. (d) Basal hydroperoxide production rate and (e) ETC inhibitor-induced hydroperoxide production rates in 7 day sham and denervated fibers treated with catalase or AACOCF3 (n = 5 M). Statistical significance determined by ordinary one-way ANOVA with Tukey’s post hoc test for each condition. Dose–response of Amplex UltraRed raw fluorescence values in response to increasing concentrations of (f) H2O2 or (g) 15(S)-HpETE in the presence of antioxidants and inhibitors at the specified concentrations. (h) Sham and denervated fibers treated with AACOCF3 and exogenous AA (n = 5 M). Statistical significance determined by ordinary two-way ANOVA with Tukey’s post hoc test. (i) Percent inhibition of basal hydroperoxide production rate in neurogenic atrophy models with catalase or AACOCF3 treatment relative to untreated controls for each model. Statistical significance determined by ordinary one-way ANOVA with Tukey’s post hoc test. For 16–19 Mo. Sod1−/−, n = 5 M total, except catalase inhibition was only performed on 4 samples and AACOCF3 inhibition was only performed on 3 samples due to limitations in the number of Oroboros O2k chambers available at the time of experimentation and thus was not powered to reach statistical significance. However, similar inhibition with AACOCF3 was observed compared to 8–9 month old Sod1−/−. End-stage SOD1G93A basal hydroperoxide production rate for vehicle and catalase or AACOCF3 inhibition was previously reported and included here for further comparison between multiple models23. *p < 0.05 versus vehicle sham or control; &p < 0.05 versus AACOCF3 sham or control; †p < 0.05 versus vehicle denervated; #p < 0.05 versus AACOCF3 denervated. All plots represent mean ± standard deviation. N.D.—No data.
