Fig 1.
Effects of MCD diet and Western diet on metabolic profile.
WT mice were fed chow diet, MCD diet for 8 weeks, or Western diet for 16 weeks, and sacrificed at 20 weeks of age. (A). Body weights; (B). Adipokines (leptin and adiponectin); (C). Fasting serum glucose and HOMA-IR index; (D). Serum triglycerides and non-esterified fatty acids (NEFA’s). Mean±SEM results are graphed. *<0.05 and **<0.005, control versus experimental diet; #<0.05 and ##<0.005, MCD versus Western diet.
Fig 2.
Effects of MCD diet and Western diet on hepatic steatosis and liver enzymes.
(A). Liver to body weight (LBW) ratio, serum aminotransferases (ALT, AST) and alkaline phosphatase (ALP) from mice fed chow, MCD or Western diets. (B). H&E staining of representative liver sections from mice (left panels) and NASH patients with mild or severe fibrosis (right panels). (C). Oil-red O staining of representative liver sections from mice (left panels) and liver lipid levels (right). (D). qRT-PCR analysis of liver genes encoding lipid metabolic enzymes. Results normalized to chow-diet fed mice and graphed as mean±SEM. *<0.05 and **<0.005, control versus experimental diet; #<0.05 and ##<0.005, MCD versus Western diet.
Fig 3.
Effects of MCD diet and Western diet on liver fibrosis and ductular reaction.
(A). Hepatic hydroxyl-proline content and qRT-PCR analysis of liver fibrosis and progenitor marker genes; (B). Liver sections from representative mice stained for markers of the fibroductular reaction (Sirius red, α-SMA, desmin, K19 and Sox-9) (left panels) and respective morphometry (right). Results normalized to chow-fed mice and graphed as mean±SEM. *<0.05 and **<0.005, control versus experimental diet; #<0.05 and ##<0.005, MCD versus Western diet.
Fig 4.
Effects of MCD diet and Western diet on liver cell death, inflammation and oxidative stress.
(A). TUNEL assay and Western blot for caspase-2 and cleaved PARP; (B). qRT-PCR of macrophage markers (F4/80 and YM-1) and TNF-α (left). Immunohistochemistry for F4/80 and YM-1: Representative photos and morphometry (right). (C). qRT-PCR analysis of anti-oxidant enzymes (top) and immunohistochemistry plus morphometry for 4-hydroxynonenal (4-HNE) in representative mice (lower). Results normalized to chow-diet fed mice and graphed as mean±SEM. *<0.05 and **<0.005, control versus experimental diet; ##<0.005, MCD versus Western diet.
Fig 5.
Effects of MCD diet and Western diet on liver cell stress and Hedgehog pathway.
(A). qRT-PCR analysis of relevant ER stress-related genes (left panel) and western blot analysis of GADD-153 (right panel); (C). Western blot analysis of the autophagy marker, LC3BII. (C). qRT-PCR analysis of Hedgehog ligands (Shh, Ihh), receptor (Patch), Hedgehog-regulated transcription factors (Gli1, Gli2), and Hedgehog-target genes (OPN, Hip-1) (left); Western blot analysis of Gli-2 (right). (D). Immunohistochemistry for osteopontin: sections from representative mice and morphometry. Results graphed as mean±SEM. *<0.05 and **<0.005, control versus experimental diet; #<0.05 and ##<0.005, MCD versus Western diet.