Well-established NAFLD/NASH mouse models successfully induce fibrosis, but struggle to recapitulate the full complexity of human disease, which often presents signs of metabolic syndrome (MS). In order to develop a more translatable NASH model that presents with MS, we fed mice a high-fat, cholesterol-rich western-diet (WD), and supplemented drinking water with fructose. A WD in combination with traditional methods to induce NASH exacerbates and facilitates the full spectrum of clinical presentations seen with human NAFLD/NASH disease, including obesity, impaired glucose tolerance and hepatic steatosis.
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Western Diet Induced NASH Model
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Western diet (WD) induced NASH model. Wild-type C57BL/6J male mice were randomly divided into 2 groups and given a standard diet (STD) or a Western diet (WD) diet consisting of high glucose and high fructose water, with various parameters measured 22 weeks after feeding. (A) Schematic representation of WD-induced NASH mouse model. (B) Body weight, (C) fasting glucose, (D) glucose tolerance test, (E) area under the curve of glucose tolerance test blood glucose level, (F) liver weight, (G) liver weight-to-body weight ratio, (H) serum ALT concentration, (I) serum total cholesterol, (J) H&E staining of liver section, (K) NAFLD (non-alcoholic fatty liver disease) activity score (NAS) were all measured in mice fed a STD or WD. Mice fed a WD showed increased body weight, fasting blood glucose, glucose tolerance test, blood glucose, liver weight, and an elevation in serum ALT and serum total cholesterol. H&E staining indicates significant steatosis, and the NAS score was significantly increased in WD-induced NASH mice. The above results indicate that mice fed a Western diet can successfully establish a NASH model. Data is represented as mean ± SEM, n = 8.
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Efficacy Evaluation of Crotedumab in a NASH Mouse Model
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Crotedumab, an anti-GCGR antibody, alleviated metabolic disorder in a WD-induced NASH mouse model. (A) Experimental schematic of NASH (non-alcoholic steatohepatitis) model establishment. Eight-week-old C57BL/6 male mice were fed a standard diet (STD) or a diet rich in cholesterol (Western diet; WD) containing 21% fat, 50% carbohydrate, and 1.5% cholesterol for 22 weeks. Blood glucose levels were analyzed and glucose tolerance tests were performed at week 13. Blood samples were collected at week 14 and analyzed for ALT and blood cholesterol. Blood samples were treated with a Crotedumab analog (synthesized in-house). (B) Body weight, (C) fasting blood glucose, (D-E) glucose tolerance test and relative values of area under the curve, (F) liver weight, (G) ratio of liver weight to body weight, (H-I) serum ALT and total cholesterol concentrations, and (J-K) H&E staining and NAS scoring of liver tissue sections were performed in mice fed a STD or WD with and without Crotedumab treatment. Results indicate that Crotedumab significantly reduced fasting blood glucose and glucose tolerance compared to mice fed a WD. Additionally, liver weight and ALT were slightly decreased, serum total cholesterol concentration was decreased, and liver tissue steatosis and ballooning were relieved. The above results indicate that administration of Crotedumab can significantly reduce the symptoms of non-alcoholic steatohepatitis induced by a Western diet. Data is represented as mean ± SEM, n = 10.
