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.

Impaired liver regeneration and lipid homeostasis in CCl4 treated WDR13 deficient mice

“…To avoid confounding effects of the absence of WDR13 per se on metabolism and the toxic effects of CCl 4 , the present study was conducted at the age of 8-10 weeks old animals when there is no onset of hyperinsulinemia and obesity. Fatty liver condition in mice has been reported upon treatment with CCl 4 along with high fat diet [42] via upregulation of PPARγ in the liver [43]. In this study we observed fatty liver phenotype in Wdr13 -/0 mice on CCl 4 treatment alone, owing to upregulation of PPAR pathway.…”

A long‐acting FGF21 alleviates hepatic steatosis and inflammation in a mouse model of non‐alcoholic steatohepatitis partly through an FGF21‐adiponectin‐IL17A pathway

“…Only two experimental studies of native FGF21 or FGF21 analogues had implicated FGF21 in the treatment of MCD diet-induced NASH (Fisher et al, 2014;Ju et al, 2016). The MCD diet-fed mouse model has a number of disadvantages, including decreased bodyweight, serum triglycerides and glucose levels (Kubota et al, 2013). By using CD-HFD to induce a type of NASH associated with obesity and a human NASH-like pathology, we could focus on the effects of long-acting FGF21 on weight loss, as PsTag600-FGF21 treatment potently induced weight loss in DIO mice (Yin et al, 2016).…”

Inhibition of Plasminogen Activator Inhibitor 1 Attenuates Hepatic Steatosis but Does Not Prevent Progressive Nonalcoholic Steatohepatitis in Mice

“…The degree of inflammation and fibrosis observed by Lee et al after feeding C57BL/6J mice a purely high‐fat diet for 10 weeks was remarkable given that mice are relatively resistant to the development of NASH with fibrosis. Moreover, it has been widely reported that even with prolonged high‐fat feeding, C57BL/6J mice develop scant or no hepatic fibrosis . As such, with the goal of identifying PAI‐1 antagonism as an antifibrotic therapy in NASH, we employed two distinct dietary models of murine NASH with fibrosis.…”