Fig. 6: Fasting is beneficial by priming for antioxidant and autophagy responses prior to APAP intoxication.

A Illustrated timeline of animal experiments with fasting 12 h prior to PBS/APAP injection and glucose injection 1 h prior to PBS/APAP injection. Glucose levels in (B + C) were measured at t = 0. Gene expression in (G) was measured at t = 2. ALT and liver damage was measured at t = 6. B Blood glucose levels of mice 1 h after 2.5 g/kg glucose injection, and of mice non-fasted or fasted overnight. Bold dotted line indicates median and weak dotted lines quartiles. C Correlation between serum ALT of mice treated with 500 mg/kg APAP for 6 h and respective blood glucose prior to APAP treatment. Crosses indicate group median +/− SEM. Statistical significance and linear fit was calculated by Pearson correlation. D Kaplan–Meier survival curve of fasted and nonfasted mice treated with APAP (6 h, 500 mg/kg APAP), n = 26 (fasted), n = 15 (non-fasted). Statistical significance was calculated using Gehan–Breslow–Wilcoxon test. E, F Representative images of H&E-stained liver sections and hemorrhage scoring of fasted or non-fasted mice treated with 500 mg/kg APAP for 6 h. Scale bars 250 µm (left), 50 µm (right). G Transcript levels of livers from nonfasted/fasted mice 2 h after glucose injection. Heatmap shows fold change to nonfasted and respective statistical significances on the right. H Fasting-induced gene ontologies of hepatocyte DEGs (left) and gene expression counts of selected DEGs (right) from overnight fasted mice analyzed by hepatocyte bulkRNA sequencing. BP biological process. Data points and/or bar graphs are mean +/− SD with n as independent biological replicates. Statistical significance was tested using Two-way ANOVA with Sidak’s multiple comparison test (B, F, G).