Figure 5

Mice fed HF/C diet plus HF/G developed hyperplastic nodules with dysplasia. 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 up to 12 months. (a) The appearance of the liver with hyperplastic nodules (B) and the whole liver of another mouse appeared to be in a hyperplastic stage (C) in mice fed HF/C diet plus HF/G and mice fed control diet, as well as representative micrographs of CD31 (D, E) and CD34 (F, G) staining at original magnification (× 400, scale bars=50 μm). Hyperplastic nodule (B) and positive staining of CD31 are indicated by arrow symbols. (b) Representative micrographs of reticular fiber, Ki67, and TUNEL staining in liver tissues at 12 months. For reticular fiber staining, immunohistochemistry images were taken at × 400 original magnification (scale bars=50 μm). For Ki67 and TUNEL (apoptosis) staining, immunohistochemistry images were taken at × 200 original magnification (scale bars=100 μm). Positive staining of reticular fiber, Ki67, and TUNEL is indicated by arrow symbols. Reduced CD31, negative CD34, much less reticular fiber, and increased Ki67 staining are in accordance with the morphologic criteria of hyperplastic nodules with dysplasia at the end stage of HF/C diet plus HF/G feeding. (c) The quantitative analysis of CD31 staining. (d) The quantitative analysis of reticular fiber staining. (e) The quantitative analysis of Ki67 staining. (f) The quantitative analysis of TUNEL staining.