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Licensed Unlicensed Requires Authentication Published by De Gruyter November 25, 2019

Development of nonalcoholic fatty liver disease model by high-fat diet in rats

  • Hijrawati Ayu Wardani , Mahardian Rahmadi , Chrismawan Ardianto , Santhra Segaran Balan , Norshafarina Shari Kamaruddin and Junaidi Khotib EMAIL logo



Nonalcoholic fatty liver disease (NAFLD) is indicated by liver steatosis without excessive alcohol use or other liver disease. Several studies have reported that metabolic syndromes such as obesity, type 2 diabetes mellitus, and dyslipidemia have a linear correlation associated with NAFLD pathophysiology. One of the characteristics of dyslipidemia in NAFLD is increase in serum triglycerides. This study aimed to develop a model of NAFLD characterized by an increase in serum triglyceride levels and histological profile of liver steatosis by high-fat diet in rats.


Twelve Wistar rats were fed with pellets enriched with 60% fat. They were housed individually, and the remaining pellets were weighted every day for intake evaluation. Blood samples were collected at day 0 and at the end of each trial period at days 7, 14, 21, and 28 for the measurement of triglyceride levels. Every animal from each group was also sacrificed for liver histopathological examination.


This study has established developing the NAFLD animal model by induction of a high-fat diet. The levels of serum triglycerides were increased from baseline 80.41 ± 12.82 to 1152.00 ± 73.62, 493.66 ± 159.98, 556.00 ± 120.79, and 489.00 ± 156.75 mg/dL at days 7, 14, 21, and 28, respectively. Liver histology also showed liver steatosis development, inflammation, and hepatocellular ballooning, which were associated with the NAFLD state.


High-fat diet in rats induced hypertriglyceridemia along with NAFLD-like liver histopathology.


The authors thank Dr. Priangga Adi Wiratama, the pathologist who technically contributed to the histology examination.

  1. Research funding: This work was supported by research grants from “Tahir Professorship Program” and PDUPT 2019 from Indonesian Ministry of Research Technology and Higher Education.

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: The authors state no conflict of interest.

  4. Ethical approval: All experiments were performed at the Animal Research Laboratory of the Faculty of Pharmacy, Universitas Airlangga (Surabaya, Indonesia), in accordance with the Guide for the Care and Use of Laboratory Animals issued by the National Institutes of Health revised in 1985. The protocol was approved by the Ethical Committee of the Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, Indonesia.


[1] Younossi ZM, Koenig AB, Abdelatif D, Fazel Y, Henry L, Wymer M. Global epidemiology of nonalcoholic fatty liver disease – meta-analytic assessment of prevalence, incidence, and outcomes. Hepatology 2016;64:73–84.10.1002/hep.28431Search in Google Scholar

[2] Farrell GC, Wong VW, Chitturi S. NAFLD in Asia – as common and important as in the West. Nat Rev Gastroenterol Hepatol 2013;10:307–18.10.1038/nrgastro.2013.34Search in Google Scholar

[3] Estes C, Anstee QM, Arias-loste MT, Bantel H, Bellentani S, Caballeria J, et al. Modeling NAFLD disease burden in China, France, Germany, Italy, Japan, Spain, United Kingdom, and United States for the period 2016–2030. J Hepatol 2018; 69:896–904.10.1016/j.jhep.2018.05.036Search in Google Scholar

[4] Almeda-Valdés P, Cuevas-Ramos D, Aguilar-Salinas CA. Metabolic syndrome and non-alcoholic fatty liver disease. Ann Hepatol 2009;8:S18–24.10.1016/S1665-2681(19)31822-8Search in Google Scholar

[5] Marchesini G, Brizi M, Bianchi G, Tomassetti S, Bugianesi E, Lenzi M, et al. Nonalcoholic fatty liver disease: a feature of the metabolic syndrome. Diabetes 2001;50:1844–50.10.1097/01.mol.0000174153.53683.f2Search in Google Scholar PubMed

[6] Wattacheril J, Chalasani N. Nonalcoholic fatty liver disease (NAFLD): is it really a serious condition? Hepatology 2012;56:1580–4.10.1002/hep.26031Search in Google Scholar PubMed PubMed Central

[7] Zivkovic AM, Sanyal AJ, German JB. Comparative review of diets for the metabolic syndrome: implications for nonalcoholic fatty liver disease. Am J Clin Nutr 2007;86:285–300.10.1093/ajcn/86.2.285Search in Google Scholar PubMed

[8] Van Herck MA, Vonghia L, Francque SM. Animal models of nonalcoholic fatty liver disease – a starter’s guide. Nutrients 2017;9:1–13.10.3390/nu9101072Search in Google Scholar PubMed PubMed Central

[9] Kleiner DE, Brunt EM, Van Natta M, Behling C, Contos MJ, Cummings OW, et al. Design and validation of a histological scoring system for nonalcoholic fatty liver disease. Hepatology 2005;41:1313–21.10.1002/hep.20701Search in Google Scholar PubMed

[10] Liang W, Menke AL, Driessen A, Koek GH, Lindeman JH. Establishment of a general NAFLD scoring system for rodent models and comparison to human liver pathology. PLoS One 2014;9:1–17.10.1371/journal.pone.0115922Search in Google Scholar PubMed PubMed Central

[11] Santhekadur PK, Kumar DP, Sanyal AJ. Preclinical models of nonalcoholic fatty liver disease. J Hepatol 2019;68:230–7.10.1016/j.jhep.2017.10.031Search in Google Scholar PubMed PubMed Central

[12] Forouhi NG, Krauss RM, Taubes G, Willett W. Dietary fat and cardiometabolic health : evidence, controversies, and consensus for guidance and colleagues. Br Med J 2018;2139:1–8.10.1136/bmj.k2139Search in Google Scholar PubMed PubMed Central

[13] Rolls BJ. The relationship between dietary energy density and energy intake. Physiol Behav 2014;97:609–15.10.1016/j.physbeh.2009.03.011Search in Google Scholar PubMed PubMed Central

[14] Ackroff K, Sclafani A. Energy density and macronutrient composition determine flavor preference conditioned by intragastric infusions of mixed diets. Physiol Behav 2006;89:250–60.10.1016/j.physbeh.2006.06.003Search in Google Scholar PubMed

[15] Ferreira JG, Tellez LA, Ren X, Yeckel CW, de Araujo IE. Regulation of fat intake in the absence of flavour. J Physiol 2012;4:953–72.10.1113/jphysiol.2011.218289Search in Google Scholar PubMed PubMed Central

[16] Ardianto C, Yonemochi N, Yamamoto S, Yang L, Takenoya F, Shioda S, et al. Opioid systems in the lateral hypothalamus regulate feeding behavior through orexin and GABA neurons. Neuroscience 2016;320:183–93.10.1016/j.neuroscience.2016.02.002Search in Google Scholar PubMed

[17] Ipsen DH, Tveden-nyborg P, Lykkesfeldt J. Dyslipidemia?: obese or not obese – that is not the question. Curr Obes Rep 2016;5:405–12.10.1007/s13679-016-0232-9Search in Google Scholar PubMed

Received: 2019-09-09
Accepted: 2019-10-23
Published Online: 2019-11-25

© 2019 Walter de Gruyter GmbH, Berlin/Boston

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