Figure 1

FK866 pre-treatment ameliorates GaIN/LPS- or ConA-induced ALF in mice and stimulates autophagy. Mice were subjected to GaIN/LPS (G/L, 600 mg/kg /0.5 μg/kg, IP) or ConA (20 mg/kg, IV) treatment with or without FK866 (FK, 10 mg/kg, IP), respectively. (a) Representative immunohistochemistry for NAMPT on paraffin-embedded liver tissue sections from control (Con) mice and G/L or ConA-induced mice. Original magnification × 400, scale bar 50 µm. n = 4 per group. (b) Western blot analysis of NAMPT protein expression in the presence or absence of FK. The data are shown as the means ± SEM. n = 4–6 per group. *P < 0.05 compared to the vehicle (Veh) group, ^# P < 0.05 compared to the Veh + G/L-7 h or Veh + ConA-6 h group. (c) The Kaplan-Meier method was used to determine the difference in survival rate after G/L or ConA challenge with or without FK. n = 11 per group. *P < 0.05 compared to the FK-treated group. (d) Quantification of serum ALT and AST levels. The data are shown as the means ± SEM. n = 6 per group. *P < 0.05 vs. the corresponding Veh group. (e) Routine histopathology was performed on formalin-fixed liver sections obtained from mice subjected to G/L or ConA challenge with or without FK. The arrows denote hepatocellular necrosis; the arrowheads denote infiltrating inflammatory cells. Original magnification × 400, scale bar 50 µm. n = 4 per group. (f) The protein expression levels of ATG7, p62 and LC3B were measured by western blot analysis. The data are shown as the means ± SEM. n = 4–6 per group. *P < 0.05 indicates significant differences. (g) To measure autophagic activity, mice received chloroquine (CQ, 60 mg/kg, 30 min, IP) prior to GaIN/LPS (G/L, 600 mg/kg /0.5 μg/kg, 3 h, IP) or ConA (20 mg/kg, 3 h, IV) challenge. Expression levels of LC3B are shown as a densitometric graph of the optical density-based data of immunoblot. The data are shown as the means ± SEM. n = 4–6 per group. *P < 0.05 vs. the corresponding Veh group, ^& P < 0.05 vs. the corresponding G/L or ConA group.