Long-term profile of serological biomarkers, hepatic inflammation, and fibrosis in a mouse model of non-alcoholic fatty liver disease
Introduction
Non-alcoholic fatty liver disease (NAFLD) can be typically classified into two subgroups: non-alcoholic fatty liver (NAFL; simple steatosis) and non-alcoholic steatohepatitis (NASH). NAFL is characterized by hepatic steatosis with no hepatocellular damage and inflammation, while NASH is characterized by the presence of hepatocellular damage, hepatic steatosis, inflammation, and fibrosis. NASH, especially with fibrosis, may lead to liver cirrhosis, which is associated with increased risk of developing hepatocellular carcinoma (HCC) (Starley et al., 2010; Vernon et al., 2011).
NAFLD mouse models are useful tools for understanding the pathogenesis and progression of NAFLD and for developing drugs for its treatment (Farrell et al., 2019; Hansen et al., 2017; Lau et al., 2017). Several NAFLD mouse models have been reported: 1) diet-induced models developed mainly through feeding the mice a high-fat diet; 2) nutrient-deficient diet-induced models developed through feeding mice a choline and/or methionine-deficient diet (with or without high-fat diet); 3) chemical-induced models developed through the injection of liver-targeted chemotoxins (with or without high-fat diet), such as carbon tetrachloride (CCl4) or streptozotocin; and 4) genetic models developed through use of high-fat or nutrient-deficient diet-fed genetic mice, such as ob/ob, db/db, and foz/foz mice (Farrell et al., 2019; Hansen et al., 2017; Lau et al., 2017).
Among these animal models, mice fed methionine and choline deficient (MCD) diets are broadly used as NAFLD mouse models. MCD diet-fed mice exhibit hepatic damage and inflammation at a relatively early stage (1–3 weeks), compared to the other animal models (Hansen et al., 2017; Lau et al., 2017). Choline deficiency disrupts liver functions, causing altered lipid metabolism, phosphatidylcholine synthesis, mitochondrial dysfunction, and reactive oxygen species-mediated DNA damage, which are closely associated with NAFLD development (Corbin and Zeisel, 2012; Sherriff et al., 2016). Furthermore, impaired S-adenosylmethionine synthesis caused by methionine deficiency leads to reduced liver metabolism, enhanced liver injury, and HCC (Anstee and Day, 2012; Mato et al., 2013).
MCD diet-fed mice show sustained reduction in body weight (BW) while on the diet. To overcome BW reduction in the MCD diet-fed mice, proteins are replaced with equivalent amounts of L-amino acids and/or high-fat, or high-sucrose diets are added to MCD diets (Farrell et al., 2019; Hansen et al., 2017; Lau et al., 2017). However, limited data are available on the time-dependent profiles of serum lipids and biochemical markers of hepatic function, hepatic inflammation, and fibrosis in the long-term in mice fed MCD diets containing L-amino acids and high-amounts of fat.
Here, we investigated the time-dependent change in serum lipids and biochemical markers of hepatic function, hepatic inflammation, and fibrosis in mice fed a NAFLD diet (choline deficient, L-amino acid-defined, high-fat diet; CDAHFD) for 12 weeks. Our data provides a better understanding of NAFLD pathogenesis.
Section snippets
Animal experiments
Animal studies were performed in accordance with the Guidelines for Animal Experiments established by the Ministry of Health, Labour and Welfare of Japan, and the National Cerebral and Cardiovascular Center Research Institute. Male C57BL/6 J mice (5-week-old) were purchased from Japan SLC (Shizuoka, Japan). They were housed in cages (maximum of five mice/cage), in a temperature-controlled room (22 °C) with a 12-h light-dark cycle. They were fed normal diet (ND) (CE-2; CLEA Japan, Inc., Tokyo,
BW change in mice
The BWs of the CDAHFD-fed mice decreased slightly during the first 2 weeks after feeding, but a time-dependent increase in BW was observed from 4 to 12 weeks. However, the changes in BW in the CDAHFD-fed mice through 1–12 weeks were not significantly different from those of mice at the start of the experiment (0 week). In the case of ND-fed mice, the BWs increased time-dependently throughout the feeding period (Fig. S1).
Lipid profiles
Serum lipid profiles of the ND- and CDAHFD-fed mice are presented in Fig. 1
Discussion
Patients with NAFL or NASH show increased serum TC, LDL-C, and TG levels but decreased serum HDL-C levels, compared to healthy subjects (Eguchi et al., 2012; Feldstein et al., 2010; Zimmermann et al., 2011). However, no substantial difference in serum lipids (TC, LDL-C, TG, and HDL-C) has been reported between NAFL and NASH (Feldstein et al., 2010; Musso et al., 2012; Zimmermann et al., 2011). Several studies have reported that the level of serum lipids decreased in patients with severe liver
Declaration of Competing Interest
The authors declare no conflict of interest.
Acknowledgment
This work was supported by theJapan Agency for Medical Research and Development (AMED) under Grant Number JP20fk0210044.
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