Figure 5.

Reduced inflammation and fibrosis in constitutive Atg5–deficient livers after chronic-plus-binge alcohol treatment. A–E:Atg7^F/F and Atg7^Δhep-ERT2 mice (18 to 24 weeks old) were injected with tamoxifen on days 3 and 4 during acclimation. After acclimation, mice were randomly divided into two groups and given a liquid diet with alcohol (EtOH-fed) or with maltose dextrin (Pair-fed) for 10 days, followed by a single gavage of alcohol (5 g/kg) or isocaloric maltose dextrin. Mice were analyzed 9 hours later. Liver sections were subjected to hematoxylin and eosin (H&E) staining (A), F4/80 stain and quantified (B and C), and Masson's trichrome stain and quantified (D and E). F: The hepatic hydroxyproline level was determined. G: Representative Western blot images of α-smooth muscle actin (Sma) protein and densitometric analysis. The density of α-Sma protein was normalized to that of β-actin. H: The mRNA expression of fibrosis-related genes in the livers of Atg5^F/F and Atg5^Δhep mice given alcohol or control diet. Expression levels of indicated genes were determined by real-time quantitative -PCR and were normalized to that of β-actin. Data are expressed as the means ± SEM fold-changes over those of Atg5^F/F mice given the pair-fed diet. n = 4 (G); n = 5 to 6 (F); n = 6 (H). ^∗P < 0.05 versus nontreated F/F control; ^†P < 0.05 versus nontreated Δhep control; ^‡P < 0.05 versus treated F/F control. Original magnification, ×200 (A, B, and D). Ctgf, connective tissue growth factor; Gfap, glial fibrillary acidic protein; Tgf, transforming growth factor; Timp1, tissue inhibitor of metallopeptidase 1.