Fig. 4.

Metabolic and proautophagic effects of α-DBI in the context of MCD. (A) Heatmap clustered by Euclidean distance of changes in liver metabolite concentrations depicted as log² (fold change) (FC) in C57BL/6 mice injected with IgG or α-DBI after MCD (Left) (n = 3–9 mice per group). All carnitines are depicted in the Right. (B–D) Expression of β-oxidation-relevant enzymes. Representative Western blot (Top) and densitometric quantification (Bottom) of CPT1A after (B) (n = 5–8 mice in each group). mRNA levels of Cpt1a (C) and Ppara (D) analyzed by qRT-PCR (n = 4–8 mice per group). (E–L) Comparison of anti-NASH effects of α-DBI in Wt and autophagy-deficient (Atg4b^−/−) mice under MCD. Representative immunoblots of autophagy markers, such as ATG4B, p62, and LC3B (E) and densitometric analysis of p62 (F) and LC3B (G). Representative histological images (H) and NAFLD activity scores (I). ALT (J) and AST (K) were measured in plasma (n = 5–8 mice per group). Expression of genes involved in inflammation (Cd68, F480, Il1b, Il6, Mcp1, Nrlp3, and Tnfa), antioxidant response (Cat, Hmox1, Nrf2, Gpx, Gsr, Sod1, and Sod2), and β-oxidation (Cpt1a, Pgc1a, and Ppara) were measured by qRT-PCR and represented in volcano plots (L) (n = 3–8 mice per group). Asterisks, arrows, and arrowheads show inflammation foci, macrosteatosis, and microsteatosis vesicular, respectively. Results are displayed as means ± SEM. For statistical analyses, P values were calculated by ANOVA test (B, D, F, G, J, and K), Kruskal–Wallis test (C and I), or two-tailed unpaired Student’s t test (L).