Carbon tetrachloride (CCl4) accelerated development of non-alcoholic fatty liver disease (NAFLD)/steatohepatitis (NASH) in MS-NASH mice fed western diet supplemented with fructose (WDF)

Abstract

Background: Multiple murine models of nonalcoholic fatty liver disease/steatohepatitis (NAFLD/NASH) have been established by using obesogenic diets and/or chemical induction. MS-NASH mouse (formally FATZO) is a spontaneously developed dysmetabolic strain that can progress from hepatosteatosis to moderate fibrosis when fed a western diet supplemented with 5% fructose (WDF). This study aimed to use carbon tetrachloride (CCl4) to accelerate and aggravate progression of NAFLD/NASH in MS-NASH mouse. Methods: Male MS-NASH mice at 8 weeks of age were fed WDF for the entire study. Starting at 16 weeks of age, CCl4 was intraperitoneally administered twice weekly at a dose of 0.2 mL/kg for 3 weeks or 0.08 mL/kg for 8 weeks. Obeticholic acid (OCA, 30 mg/kg, QD) was administered in both MS-NASH and C57Bl/6 mice fed WDF and treated with CCl4 (0.08 mL/kg).Results: WDF enhanced obesity and hepatosteatosis, as well as induced moderate fibrosis in MS-NASH mice similar to previous reports. Administration of CCl4 accelerated liver fibrosis with increased bridging and liver hydroxyproline contents, but had no significant impact on liver steatosis and lipid contents. High dose CCl4 caused high mortality and dramatic elevation of ALT and ASL, while low dose CCl4 resulted in a moderate elevation of ALT and AST with low mortality. Compared to C57BI/6 mice with WDF and CCl4 (0.08 mL/kg), MS-NASH mice had more prominent hepatosteatosis and fibrosis. OCA treatment significantly lowered liver triglycerides, steatosis and fibrosis in both MS-NASH and C57Bl/6 mice fed WDF with CCl4 treatment. Conclusions: CCl4 reduced induction time and exacerbated liver fibrosis in MS-NASH mice on WDF, proving a superior NASH model with more prominent liver pathology, which has been used favorably in pharmaceutical industry for testing novel NASH therapeutics.

Mouse models of nonalcoholic fatty liver disease (NAFLD): pathomechanisms and pharmacotherapies

“…Repeated administration of CCl 4 to HFD-fed obese mice successfully induced chronic oxidative stress, triggered inflammation and led to liver fibrosis [92]. Notably, under feeding with Western diet (WD) supplemented with 5% fructose (WDF), CCl 4 reduced the induction time and aggravated liver fibrosis in FATZO mice [93]. In comparison to CDAA-treated mice, additional treatment with a single dose of CCl 4 resulted in not merely hepatic lipid deposition but also peri-hepatocellular fibrosis [94].…”

Establishment of animal models for NASH and utilization for mining associated gut microbiota

“…21 A low weekly dose of intraperitoneal CCl 4 acts as a NASH accelerator in a western diet-fed animal model. 22 CCl 4 can be converted to trichloromethyl (•CCl 3 ) and trichloromethyl peroxyl (•CCl 3 O 2 ) radicals by the cytochrome P450 enzyme. These radicals damage the tissue by increasing oxidative stress and lipid peroxidation.…”

Automated Whole Slide Image Analysis for a Translational Quantification of Liver Fibrosis

“…Accurate assessment of brosis is paramount to determine the severity of chronic liver diseases, such as non-alcoholic steatohepatitis (NASH), as it primarily drives mortality in patients [1][2][3][4]. In preclinical studies, brosis amount is evaluated from the biochemical assay of hydroxyproline (HYP) content, providing a continuous measurement of collagen content and correlating with the severity of liver brosis [5][6][7][8]. In parallel, liver brosis is also evaluated by means of histological scoring systems which are applied for both preclinical studies and clinical investigations from human biopsy [9,10].…”