Figure 4

Mice fed HFC diet plus HF/G developed hepatic fibrosis. C57BL/6J mice were fed a control diet or high-fat/-calorie diet plus high fructose/glucose in drinking water (HF/C diet plus HF/G) for 12 months. (a) Representative micrographs and semi-quantification (b) of immunohistochemical staining of collagen I in mouse liver at 5, 9, and 12 months. The positive signal was developed with DAB (in brown color indicated by arrow symbols). Representative images were recorded at original magnification (× 400), and scale bars=50 μm. (c) Representative micrographs of Masson’s trichrome staining of liver tissues at 5, 9, and 12 months. Hepatic fibrosis is indicated by arrow symbols. Images were recorded at original magnification (× 200). Scale bars=100 μm. The increase in hydroxyproline content overtime is consistent with progressive fibrotic deposition in the parenchyma and the portal triads. (d) The quantitative analysis of Masson’s trichrome staining. (e) mRNA level of α-SMA in mouse liver at 5, 9, and 12 months. (f, g) mRNA levels of procollagen type I (COL1A1 and COL1A2) in mouse liver at 5, 9, and 12 months. (h) Liver hydroxyproline content (ng/mg tissue) at 5, 9, and 12 months. Data are expressed as the means±s.e.m. Number of mice in control group at 5 and 9 months=3; number of mice in control group at 12 months=4; number of mice in HF/C diet plus HF/G group at 5 and 9 months=5; and number of mice in HF/C diet plus HF/G group at 12 months=10. *P<0.05 and **P<0.01 compared to mice fed control diet at corresponding time points. Figure legends are the same in panel e, f, g and h.