Figure 2.

Increasing dietary MCFA ratio attenuated NASH development in mice. To investigate the protective effects of MCFAs on NASH development, paraffin-embedded mouse liver sections were stained with H&E and Sirius Red to evaluate hepatic steatosis (A) and fibrosis (B) respectively. Hepatic TG content (C), collagen I protein and hydroxyproline (D) levels were also quantified using a commercial assay kit and dot blotting assay. Hepatic insulin resistance, autophagic flux, ER stress, and apoptosis were analyzed using Western blotting. For insulin resistance, the hepatic phospho-Akt (Ser473) level in mice injected with or without I.P. insulin (0.5 IU/kg body weight, 30 min) were analyzed. Representative immunoblots and densitometric quantification of phospho-Akt (Ser473) (E), and the calculated insulin-stimulated Akt phosphorylation fold change results (F) are presented. Representative SQSTM1/p62, LC3, CHOP, and cleaved caspase 3 immunoblots and densitometric quantifications (G) indicate that increasing MCFA ratio attenuated HFD-induced hepatic autophagy impairment, ER stress, and cell apoptosis. For densitometric analyses of Western blotting data, β-actin and Akt (for Akt phosphorylation analysis) were used as the loading controls. Values are mean ± SEM (n = 6). *, ^#(vs. CTD fed mice without insulin injection), and ^†(vs. SDHFD fed mice without insulin injection) indicate statistical significance, P < 0.05. Full-length blots are presented in Supplementary Figure S2.