Figure 8

Fibrosis markers and injury- and hepatic apoptosis-related parameters in db/db mice after 12 weeks. (a) Plasma aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels also were measured in each murine methionine choline deficient (MCD) diet-fed db/db mouse group after 12 weeks of treatment. Mean±s.e. results from each group (n=5/group) (^*P<0.05). (b) The number of inflammatory foci per 200 × field was counted on sections from each mouse. Mean±s.e. data from each group (n=5/group) are displayed (^*P<0.05). (c) The mRNA levels of tumor necrosis factor-alpha (TNF-α), TGFβ-1, collagen, α-smooth muscle actin (α-SMA) and tissue inhibitor of metalloproteinases (TIMP-1) also were evaluated by quantitative real-time PCR analysis from each MCD diet-fed db/db mouse group at 12 weeks. Results were normalized to glucuronidase (GUS) expression and then expressed as fold changes relative to gene expression in MCD diet-fed control db/db mice. Mean±s.e. data from each group (n=5/group) (^*P<0.05, ^**P<0.01). (d) Morphometric analysis of Sirius Red-stained sections from each MCD diet-fed group (n=5/group) at 12 weeks. Results are expressed as percentage of section staining (+) for Sirius Red (^*P<0.05). (e) Hepatic Bcl-2 and Ref-1 levels were evaluated by immunoblot analysis of livers from three mice/group after 12 weeks. (f) Apoptosis of hepatocytes was evaluated by TUNEL staining of liver sections after 12 weeks of treatment. TUNEL-positive hepatocytes were counted in three randomly selected fields/section ( × 100 magnification) from each MCD diet-fed group (n=5/group) at 12 weeks. Results are expressed as percentage of nuclei staining (+) for TUNEL (^*P<0.05).