Novel non‐alcoholic steatohepatitis model with histopathological and insulin‐resistant features

Abstract

Although several non-alcoholic steatohepatitis (NASH) models have been reported to date, few of these models fully reflect the histopathology and pathophysiology of human NASH. The aim of this study was to establish a novel NASH model by feeding a high-fat (HF) diet and administering both carbon tetrachloride (CCl 4 ) and the Liver X receptor agonist T0901317. Male C57BL/6J mice were divided into four groups (each n ¼ 5): HF, HF þ CCl 4 , HF þ T0901317, and the novel NASH model (HF þ CCl 4 þ T0901317). CCl 4 (0.1 mL/kg) and T0901317 (2.5 mg/ kg) were intraperitoneally administered four times and five times, respectively. The livers of the novel NASH model group presented a whitish colour. The serum levels of TNF-a and IL-6 were significantly increased in the novel NASH model group, and mice in this group exhibited histopathological features and insulin resistance reflective of NASH, i.e., macrovesicular hepatic steatosis, ballooning hepatocytes, Mallory-Denk bodies, lobular inflammation and fibrosis. The novel NASH model group presented significantly upregulated expression levels of mRNAs related to lipogenesis, oxidative stress, fibrosis and steatosis and significantly downregulated expression levels of mRNAs related to triglyceride export. We successfully established a novel experimental NASH model that exhibits similar histopathology and pathophysiology to human NASH.Key words: insulin resistance, Liver X receptor, mouse model, NAFLD activity score, non-alcoholic steatohepatitis Metabolic syndrome has been increasing worldwide concomitant with the increased prevalence of obesity, diabetes mellitus, and hyperlipidemia. Non-alcoholic fatty liver disease (NAFLD) was recently recognized as a hepatic manifestation of metabolic syndrome. A high-fat (HF) diet model is also widely used for the basic study of NASH because it causes obesity and insulin resistance. 15 In the HF diet models, long-term feeding of a HF diet leads to steatohepatitis that resembles that observed in human NASH. 16 However, this model is not suitable for researching the pathogenesis of NASH because a long time is required for the development of NASH symptoms and because the hepatic fibrosis in the model animals is weaker than that found in human NASH. 16Therefore, the establishment of a novel animal model that exhibits features similar to those of human NASH is necessary for the basic study of NASH. We aimed to develop a novel NASH model by feeding an HF diet and administering both carbon tetrachloride (CCl 4 ) and T0901317, which resulted in a combination of multiple important factors, including overnutrition, oxidative stress and lipogenesis. The administration of CCl 4 is known to induce chronic oxidative stress and liver injury in mice, and CCl 4 has been used to create acute hepatitis animal models. 17 An analysis of histopathological findings has revealed that CCl 4 leads to inflammation and fibrosis but not steatosis or ballooning. The liver X receptor (LXR) is a nuclear receptor that plays a key role in regulating cholesterol, fatty ac...

Modeling alcoholic liver disease in a human Liver-Chip

“…Our data also show that the ALD Liver-Chip picked up the impact of ethanol treatment on genes involved in the metabolism of alanine, aspartate, and glutamate. In line with previous data shown the link between FLD and DNA damage in hepatocytes 18 , our analysis revealed upregulation of POLE and POLD2 61 , both involved in DNA replication and repair, as well as of RAD51 and FANCB, which are expressed at DNA damage sites and implicated in homologous recombination 62,63 (Fig. 3e).…”

Evaluation of safety for hepatectomy in a novel mouse model with nonalcoholic-steatohepatitis

“…NASH mice were fed an HF diet (60 kcal% fat; D12492, Research Diets, Inc., New Brunswick, NJ, United States) for 4 wk, intraperitoneally injected with CCl 4 (Wako Pure Chemical Industries, Ltd., Osaka, Japan) twice a week for the final 2 wk, and intraperitoneally injected with T0901317 (Cayman Chemical Co., Ann Arbor, MI, United States) solubilized in DMSO for the final 5 d. The CCl 4 dose was 0.1 mL/kg, and the T0901317 dose was 2.5 mg/kg[ 28 ].…”

“…The normal liver mice have been not added any reagent and the histology and pathology have been not change. In 30% PH and 70% PH of the NASH liver group, liver specimens were evaluated by an experienced pathologist in a blinded fashion, the histology and pathology finding in the NASH severity of each groups have resulted in no difference in the NAFLD activity scores[ 28 ]. All mice received the hepatectomy 48 h after the final administration of CCl 4 and T0901317.…”

“…For the same reasons, the effect of hepatectomy on NASH liver has not been clearly elucidated in previous reports. We established a novel experimental NASH model that indicates similar histopathological and pathophysiological characteristics as those of NASH in humans[ 28 ]. The aim of this study was to investigate whether a difference in liver resection volume in a novel NASH model influences surgical outcomes.…”

Molecular Characterization of Microvesicular and Macrovesicular Steatosis Shows Widespread Differences in Metabolic Pathways

“…To date, analysis of hepatic steatosis has been based on histological analysis, whole tissue homogenate analysis, or in vitro studies (Aljomah et al, 2015;Chen et al, 2014;Fujimoto et al, 2004;Guillén et al, 2009;Owada et al, 2018;Zhou et al, 2012). Here, we present a LCM-based method to assess the differences between microsteatotic and macrosteatotic compartments and outline the principal metabolic events behind steatosis development in a mouse model of liver steatosis.…”