Journal of Gastroenterology

, Volume 41, Issue 10, pp 971–980 | Cite as

Cholesterol-fed rabbit as a unique model of nonalcoholic, nonobese, non-insulin-resistant fatty liver disease with characteristic fibrosis

  • Mosaburo Kainuma
  • Makoto Fujimoto
  • Nobuyasu Sekiya
  • Koichi Tsuneyama
  • Chunmei Cheng
  • Yasuo Takano
  • Katsutoshi Terasawa
  • Yutaka Shimada



The number of patients suffering from metabolic syndrome is increasing rapidly. Metabolic syndrome causes severe pathological changes in various organs, including the liver, and its main phenotype is nonalcoholic fatty liver disease (NAFLD). NAFLD has a broad spectrum ranging from simple fatty change to severe steatohepatitis with marked fibrosis. Recently, several experimental animal models for NAFLD have been proposed. However, most were established by rather artificial conditions such as genetic alteration. In the present study, we tried to establish a unique animal model mimicking some of the physiopathological features of NAFLD using high-cholesterol-fed rabbits.


Male rabbits fed with standard rabbit food containing 1% cholesterol for 8 weeks and 12 weeks were compared to controls (six rabbits/group). The weight of food was strictly restricted to 100 g/rabbit per day.


Body weights and fasting plasma insulin levels showed no significant differences among the groups. In contrast, characteristic fine fibrosis was extended from perivenular to pericellular areas, and microvesicular fatty change with ballooning degeneration was observed in perivenular areas in livers of the cholesterol-fed rabbits. Increase of serum cholesterol level, activation of hepatic stellate cells, and exposure to oxidative stress were also recognized.


Cholesterol-fed rabbits share several physiopathological features of NAFLD. Because this model did not show insulin resistance or obesity, it may be useful for elucidating the mechanism of NAFLD related mainly to hyperlipidemia.

Key words

nonalcoholic fatty liver disease hepatic stellate cell hyperlipidemia oxidative stress adipocyte differentiation-related protein 


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  1. 1.
    Hegele, RA, Pollex, RL 2005Genetic and physiological insights into the metabolic syndromeAm J Physiol Regul Integr Comp Physiol2896639Google Scholar
  2. 2.
    Bestermann, W, Houston, MC, Basile, J, Egan, B, Ferrario, CM, Lackland, D,  et al. 2005Addressing the global cardiovascular risk of hypertension, dyslipidemia, diabetes mellitus, and the metabolic syndrome in the southeastern United States, part II: treatment recommendations for management of the global cardiovascular risk of hypertension, dyslipidemia, diabetes mellitus, and the metabolic syndromeAm J Med Sci329292305PubMedCrossRefGoogle Scholar
  3. 3.
    James, OFW, Day, CP 1998Non-alcoholic steatohepatitis (NASH): a disease of emerging identity and importanceJ Hepatol29495501PubMedCrossRefGoogle Scholar
  4. 4.
    Clark, JM, Brancati, FL, Diehl, AM 2002Nonalcoholic fatty liver diseaseGastroenterology122164957PubMedCrossRefGoogle Scholar
  5. 5.
    Ruhl, CE, Everhart, JE 2003Determinants of the association of overweight with elevated serum alanine aminotransferase activity in the Unites StatesGastroenterology124719PubMedCrossRefGoogle Scholar
  6. 6.
    Bellentani, S, Saccoccio, G, Masutti, F, Crocè, LS, Brandi, G, Sasso, F,  et al. 2000Prevalence of and risk factors for hepatic steatosis in northern ItalyAnn Intern Med1321127PubMedGoogle Scholar
  7. 7.
    Nomura, H, Kashiwagi, S, Hayashi, J, Kajiyama, W, Tani, S, Goto, M 1988Prevalence of fatty liver in a general population of Okinawa JapanJpn J Med271429PubMedGoogle Scholar
  8. 8.
    Matteoni, CA, Younossi, ZM, Gramlich, T, Boparai, N, Liu, YC, McCullough, AJ 1999Nonalcoholic fatty liver disease: a spectrum of clinical and pathological severityGastroenterology11614139PubMedCrossRefGoogle Scholar
  9. 9.
    Powell, EE, Cooksley, WGE, Hanson, R, Searle, J, Halliday, JW, Powell, LW 1990The natural history of nonalcoholic steatohepatitis: a follow-up study of forty-two patients for up to 21 yearsHepatology117480PubMedGoogle Scholar
  10. 10.
    Bacon, BR, Farahvash, MJ, Janney, CG, Neuschwander-Tetri, BA 1994Nonalcoholic steatohepatitis: an expanded clinical entityGastroenterology10711039PubMedGoogle Scholar
  11. 11.
    Sumida, Y, Nakashima, T, Yoh, T, Furutani, M, Hirohama, A, Kakisaka, Y,  et al. 2003Serum thioredoxin levels as a predictor of steatohepatitis in patients with nonalcoholic fatty liver diseaseJ Hepatol38328PubMedCrossRefGoogle Scholar
  12. 12.
    Seki, S, Kitada, T, Yamada, T, Sakaguchi, H, Nakatani, K, Wasaka, K 2002In situ detection of lipid peroxidation and oxidative DNA damage in nonalcoholic fatty liver diseaseJ Hepatol375662PubMedCrossRefGoogle Scholar
  13. 13.
    Day, CP, James, OFW 1998Steatohepatitis: a tale of two “hits”?Gastroenterology1148425PubMedCrossRefGoogle Scholar
  14. 14.
    Weltman, MD, Farrell, GC, Hall, P, Ingelman-Sundberg, M, Liddle, C 1998Hepatic cytochrome P450 2E1 is increased in patients with non-alcoholic steatohepatitisHepatology2712833PubMedCrossRefGoogle Scholar
  15. 15.
    Leclercq, IA, Farrell, GC, Schriemer, R, Robertson, GR 2002Leptin is essential for the hepatic fibrogenic response to chronic liver injuryJ Hepatol3720613PubMedCrossRefGoogle Scholar
  16. 16.
    Koteish, A, Diehl, AM 2001Animal models of steatosisSemin Liver Dis2189104PubMedCrossRefGoogle Scholar
  17. 17.
    George, J, Pera, N, Phung, N, Leclercq, I, Yun, Hou J, Farrell, G 2003Lipid peroxidation, stellate cell activation and hepatic fibrogenesis in a rat model of chronic steatohepatitisJ Hepatol3975664PubMedCrossRefGoogle Scholar
  18. 18.
    Lieber, CS, Leo, MA, Mak, KM, Xu, Y, Cao, Q, Ren, C,  et al. 2004Model of nonalcoholic steatohepatitisAm J Clin Nutr795029PubMedGoogle Scholar
  19. 19.
    Fan, JG, Zhong, L, Xu, ZJ, Tia, LY, Ding, XD, Li, MS,  et al. 2003Effects of low-calorie diet on steatohepatitis in rats with obesity and hyperlipidemiaWorld J Gastroenterol920459PubMedGoogle Scholar
  20. 20.
    Finking, G, Hanke, H 1997Nikolaj Nikolajewitsch Antischkow (1885–1964) established the cholesterol-fed rabbit as a model for atherosclerosis researchAtherosclerosis13517PubMedCrossRefGoogle Scholar
  21. 21.
    Gupta, PP, Tandon, HD, Ramalingaswami, V 1976Cirrhosis of the liver in rabbits induced by a high cholesterol diet: an experimental modelIndian J Med Res64151626PubMedGoogle Scholar
  22. 22.
    Buyssens, N, Kockx, MM, Herman, AG, Lazou, JM, Van den Berg, K, Wisse, E,  et al. 1996Centrolobular liver fibrosis in the hypercholesterolemic rabbitHepatology2493946PubMedCrossRefGoogle Scholar
  23. 23.
    Wanless, IR, Belgiorno, J, Huet, PM 1996Hepatic sinusoidal fibrosis induced by cholesterol and stilbesterol in the rabbit: 1. morphology and inhibition of fibrogenesis by dipyridamoleHepatology2485564PubMedCrossRefGoogle Scholar
  24. 24.
    Sekiya, N, Tanaka, N, Itoh, T, Shimada, Y, Goto, H, Terasawa, K 1999Keishi-bukuryo-gan prevents the progression of atherosclerosis in cholesterol-fed rabbitPhytother Res131926PubMedCrossRefGoogle Scholar
  25. 25.
    Yoshiji, H, Kuriyama, S, Yoshii, J, Ikenaka, Y, Noguchi, R, Nakatani, T,  et al. 2001Angiotensin-II type 1 receptor interaction is a major regulator for liver fibrosis development in ratsHepatology3474550PubMedGoogle Scholar
  26. 26.
    Saadeh, S, Younossi, ZM, Remer, EM, Gramlich, T, Ong, JP, Hurley, M,  et al. 2002The utility of radiological imaging in nonalcoholic fatty liver diseaseGastroenterology12374550PubMedCrossRefGoogle Scholar
  27. 27.
    Washington, K, Wright, K, Shyr, Y, Hunter, EB, Olson, S, Raiford, DS 2000Hepatic stellate cell activation in nonalcoholic steatohepatitis and fatty liverHum Pathol318228PubMedCrossRefGoogle Scholar
  28. 28.
    Kasai, H 1997Analysis of a form of oxidative DNA damage, 8-hydroxy-2′-deoxyguanosine, as a marker of cellular oxidative stress during carcinogenesisMutat Res38714763PubMedCrossRefGoogle Scholar
  29. 29.
    Kleiner, DE, Brunt, EM, Van Natta, M, Behling, C, Contos, MJ, Cummings, OW,  et al. 2005Design and validation of a histological scoring system for nonalcoholic fatty liver diseaseHepatology41131321PubMedCrossRefGoogle Scholar
  30. 30.
    Fromenty, B, Pessayre, D 1995Inhibition of mitochondrial beta-oxidation as a mechanism of hepatotoxicityPharmacol Ther6710154PubMedCrossRefGoogle Scholar
  31. 31.
    Chitturi, S, Abeygunasekera, S, Farrell, GC, Holmes-Walker, J, Hui, JM, Fung, C,  et al. 2002NASH and insulin resistance: insulin hypersecretion and specific association with the insulin resistance syndromeHepatology353739PubMedCrossRefGoogle Scholar
  32. 32.
    Pagano, G, Pacini, G, Musso, G, Gambino, R, Mecca, F, Depetris, N,  et al. 2002Non-alcoholic steatohepatitis, insulin resistance, and metabolic syndrome: further evidence for an etiologic associationHepatology3536772PubMedCrossRefGoogle Scholar
  33. 33.
    Sanyal, AJ, Campbell-Sargent, C, Mirshahi, F, Rizzo, WB, Contos, MJ, Sterling, RK,  et al. 2001Non-alcoholic steatohepatitis: association of insulin resistance and mitochondrial abnormalitiesGastroenterology120118392PubMedCrossRefGoogle Scholar
  34. 34.
    Rinella, ME, Green, RM 2004The methionine–choline deficient dietary model of steatohepatitis does not exhibit insulin resistanceJ Hepatol404751PubMedCrossRefGoogle Scholar
  35. 35.
    Chen, A, Zhang, L, Xu, J, Tang, J 2002The antioxidant (–)-epigallocatechin-3-gallate inhibits activated hepatic stellate cell growth and suppresses acetaldehyde-induced gene expressionBiochem J368695704PubMedCrossRefGoogle Scholar
  36. 36.
    Pietrangelo, A 1996Metals, oxidative stress, and hepatic fibrogenesisSemin Liver Dis161330PubMedCrossRefGoogle Scholar
  37. 37.
    Stärkel, P, Sempoux, C, Leclercq, I, Herin, M, Deby, C, Desager, JP,  et al. 2003Oxidative stress, KLF6 and transforming growth factor-β up-regulation differentiate non-alcoholic steatohepatitis progressing to fibrosis from uncomplicated steatosis in ratsJ Hepatol3953846PubMedCrossRefGoogle Scholar
  38. 38.
    Matsuoka, M, Tsukamoto, H 1990Stimulation of hepatic lipocyte collagen production by Kupffer cell-derived transforming growth factor β: implication for a pathogenetic role in alcoholic liver fibrogenesisHepatology11599605PubMedGoogle Scholar
  39. 39.
    Nakatsukasa, H, Nagy, P, Everts, RP, Hsia, CC, Marsden, E, Thorgeirsson, SS 1990Cellular distribution of transforming growth factor-β1 and procollagen types I, III, and IV transcripts in carbon tetrachloride-induced rat liver fibrosisJ Clin Invest85183343PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Tokyo 2006

Authors and Affiliations

  • Mosaburo Kainuma
    • 1
  • Makoto Fujimoto
    • 1
    • 2
  • Nobuyasu Sekiya
    • 3
  • Koichi Tsuneyama
    • 2
    • 4
  • Chunmei Cheng
    • 5
  • Yasuo Takano
    • 2
  • Katsutoshi Terasawa
    • 4
    • 6
  • Yutaka Shimada
    • 1
    • 4
  1. 1.Department of Japanese Oriental Medicine, Faculty of MedicineUniversity of ToyamaToyamaJapan
  2. 2.First Department of Pathology, Faculty of MedicineUniversity of ToyamaToyamaJapan
  3. 3.Department of Frontier Japanese Oriental MedicineGraduate School of Medicine, Chiba UniversityChibaJapan
  4. 4.21st Century COE ProgramUniversity of ToyamaToyamaJapan
  5. 5.Allergy and Clinical ImmunologyUniversity of CaliforniaDavisUSA
  6. 6.Department of Japanese Oriental MedicineGraduate School of Medicine, Chiba UniversityChibaJapan

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