title: A high-fat diet and multiple administration of carbon tetrachloride induces liver injury and pathological features associated with non-alcoholic steatohepatitis in mice.
abstract: The aim of the present study was to establish a progressive steatohepatitis mouse model because few reported animal models of non-alcoholic steatohepatitis (NASH) show the progression from fatty liver to steatohepatitis. C57BL/6N mice were fed a high-fat diet (HFD) to develop obesity and were either administered carbon tetrachloride (CCl4 ) eight times (0.05 mL/kg, s.c., once, followed by 0.1 mL/kg, s.c., seven times) or not. Serum parameters and hepatic histopathology were examined. In a separate experiment, CCl4 was administered subcutaneously from 0 to eight times to HFD-fed obese mice to investigate progressive changes. Markers of oxidative stress, inflammation and apoptosis, as well as histopathological changes in the liver, were analysed. The HFD-fed obese mice showed fatty liver but not steatohepatitis. In contrast, HFD-fed mice administered CCl4 eight times showed histopathological features of steatohepatitis (fatty liver, inflammation, hepatocellular ballooning and fibrosis) and increased serum alanine aminotransferase levels. However, the multiple administration of CCl4 to obese mice reduced the ratio of reduced glutathione to oxidized glutathione, superoxide dismutase activity and mitochondrial DNA copy number, leading to the development of chronic oxidative stress, increased numbers of apoptotic cells and increased levels of both tumour necrosis factor-α and transforming growth factor-β mRNA. The resulting inflammation led to increased hydroxyproline content in the liver and fibrosis. The present study demonstrates that multiple administration of CCl4 to HFD-fed obese mice induces chronic oxidative stress that triggers inflammation and apoptosis and leads to the development of fibrosis in the liver, resulting in progression from fatty liver to steatohepatitis. This murine model will be useful in the research of hepatic disorders.
authors.affiliation: Yakult Central Institute for Microbiological Research, Tokyo, Japan. norihiro-kubota@yakult.co.jp
authors.forename: Norihiro
authors.initials: N
authors.lastname: Kubota
authors.forename: Shoichi
authors.initials: S
authors.lastname: Kado
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Kano
authors.forename: Norie
authors.initials: N
authors.lastname: Masuoka
authors.forename: Yuriko
authors.initials: Y
authors.lastname: Nagata
authors.forename: Toshihide
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Kouji
authors.initials: K
authors.lastname: Miyazaki
authors.forename: Fumiyasu
authors.initials: F
authors.lastname: Ishikawa
mesh_terms: D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002251:Carbon Tetrachloride / Q000009:adverse effects*; D056915:DNA Copy Number Variations / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D059305:Diet, High-Fat / Q000009:adverse effects*; D004195:Disease Models, Animal; D005234:Fatty Liver / Q000139:chemically induced* / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D005355:Fibrosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D005978:Glutathione / Q000235:genetics / Q000378:metabolism; D019803:Glutathione Disulfide / Q000235:genetics / Q000378:metabolism; D007249:Inflammation / Q000209:etiology / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008107:Liver Diseases / Q000378:metabolism / Q000473:pathology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D065626:Non-alcoholic Fatty Liver Disease; D009765:Obesity / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D004272:DNA, Mitochondrial; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D002251:Carbon Tetrachloride; D013482:Superoxide Dismutase; D000410:Alanine Transaminase; D005978:Glutathione; D019803:Glutathione Disulfide