Basic Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Sep 7, 2019; 25(33): 4904-4920
Published online Sep 7, 2019. doi: 10.3748/wjg.v25.i33.4904
Towards a standard diet-induced and biopsy-confirmed mouse model of non-alcoholic steatohepatitis: Impact of dietary fat source
Michelle L Boland, Denise Oró, Kirstine S Tølbøl, Sebastian T Thrane, Jens Christian Nielsen, Taylor S Cohen, David E Tabor, Fiona Fernandes, Andrey Tovchigrechko, Sanne S Veidal, Paul Warrener, Bret R Sellman, Jacob Jelsing, Michael Feigh, Niels Vrang, James L Trevaskis, Henrik H Hansen
Michelle L Boland, Taylor S Cohen, David E Tabor, Fiona Fernandes, Andrey Tovchigrechko, Paul Warrener, Bret R Sellman, James L Trevaskis, Cardiovascular, Renal and Metabolic Diseases, MedImmune, Gaithersburg, MD 20878, United States
Michelle L Boland, Denise Oró, Kirstine S Tølbøl, Sebastian T Thrane, Jens Christian Nielsen, Sanne S Veidal, Jacob Jelsing, Michael Feigh, Niels Vrang, Henrik H Hansen, Pharmacology, Gubra, Hørsholm DK-2970, Denmark
Author contributions: Boland ML, Cohen TS, Warrener P, Sellman BR, Feigh M, Vrang N, Trevaskis JL, and Hansen HH designed and coordinated the study; Boland ML, Oró D, Tølbøl KS, Thrane ST, Nielsen JC, Tabor DE, and Fernandes F performed the experiments, acquired and analyzed data; Boland ML, Cohen TS, Tabor DE, Fernandes F, Oró D, Tølbøl KS, Thrane ST, Nielsen JC, Tovchigrechko A, Veidal SS, Feigh M, Jelsing J, Vrang N, Trevaskis JL, and Hansen HH interpreted the data; Boland ML, Jelsing J, Trevaskis JL, and Hansen HH wrote the manuscript; all authors approved the final version of the article.
Supported by the Innovation Fund Denmark, Tølbøl KS, No. 5016-00168B.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board at MedImmune and Gubra.
Institutional animal care and use committee statement: All animal experiments conformed to the internationally accepted principles for the care and use of laboratory animals (licence No. 2013-15-2934-00784, The Animal Experiments Inspectorate, Denmark; protocol no. MI-17-0005, The Institutional Animal Care and Use Committee at MedImmune, Gaitherburg, MD, United States).
Conflict-of-interest statement: Michelle L. Boland and James L. Trevaskis were previously employed by MedImmune, LLC. Taylor S. Cohen, David Tabor, Fiona Fernandes, Andrey Tovchigrechko, Paul Warrener, and Bret R. Sellman are employed by MedImmune LLC. All other authors have nothing to disclose.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and theuse is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Henrik H Hansen, PhD, Senior Scientist, Pharmacology, Gubra, Hørsholm Kongevej 11B, Hørsholm DK-2970, Denmark. hbh@gubra.dk
Telephone: +45-31-522-651
Received: April 26, 2019
Peer-review started: April 26, 2019
First decision: May 24, 2019
Revised: June 28, 2019
Accepted: July 19, 2019
Article in press: July 19, 2019
Published online: September 7, 2019
Processing time: 134 Days and 21 Hours
ARTICLE HIGHLIGHTS
Research background

Non-alcoholic steatohepatitis (NASH) is an obesity-associated liver disease with marked unmet medical need. Various diet-induced obese animal models of NASH have been employed in preclinical research, target discovery and drug development. The trans-fat containing amylin liver NASH (AMLN) diet, high in fat, fructose and cholesterol, has been widely used in ob/ob and C57BL/6J mice for reliably inducing metabolic and liver histopathological changes recapitulating hallmarks of NASH.

Research motivation

A recent ban on trans-fats as food additive has prompted the development of a trans-fat free high-fat diet capable of promoting a compatible level of disease in ob/ob and C57BL/6J mice.

Research objectives

The present study aimed to develop and characterize a liver biopsy-confirmed obese mouse model of NASH based on an isocaloric palmitic acid-enriched diet with a nutrient composition similar to the AMLN diet.

Research methods

Male ob/ob mice were fed AMLN diet or a modified AMLN diet with trans-fat (Primex shortening) substituted by equivalent amounts of palm oil [Gubra Amylin NASH, (GAN) diet] for 8, 12 and 16 wk. In addition, C57BL/6J mice were fed AMLN or GAN diet for 28 wk. AMLN and GAN diets were isocaloric (40% fat kcal; 10% sucrose, 22% fructose, 2% cholesterol). Disease phenotyping included metabolic, liver biochemical/histopathological/transcriptomics as well as gut microbiome analyses.

Research results

In ob/ob mice, the GAN diet was more obesogenic and adipogenic compared to the AMLN diet. Whereas the GAN diet promoted impaired oral glucose tolerance in ob/ob mice, the AMLN diet had no effect on glucose regulation. The GAN and AMLN diets induced similar severity of liver biopsy-confirmed steatosis, lobular inflammation, hepatocyte ballooning and fibrotic lesions. In addition, hepatic transcriptome and gut microbiome changes were similar in ob/ob mice fed the GAN and AMLN diet. Also, C57BL/6J mice fed the GAN and AMLN developed a similar histological phenotype of mild to moderate fibrotic NASH.

Research conclusions

Substitution of trans-fat (Primex in the AMLN diet) with saturated fat (palm oil in the GAN diet) promotes a consistent phenotype of biopsy-confirmed fibrotic NASH in both ob/ob and C57BL/6J mice.

Research perspectives

GAN diet-based ob/ob and C57BL/6J mouse models of biopsy-confirmed NASH are applicable for preclinical characterization of novel NASH treatments.