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Comparative study of fatty liver induced by methionine and choline-deficiency in C57BL/6N mice originating from three different sources

Abstract

Non-alcoholic fatty liver disease (NAFLD) is believed to be the most prevalent liver disease worldwide and a major cause of chronic liver injury. It is characterized by lipid accumulation in the absence of significant alcohol consumption and frequently progresses to steatohepatitis, liver fibrosis, and hepatocellular carcinoma. Although many studies have been conducted to better understand NAFLD since it was first recognized, there are still many gaps in knowledge of etiology, prognosis, prevention and treatment. Methionine-choline deficient (MCD) diet, a well-established experimental model of NAFLD in rodents, rapidly and efficiently produces the clinical pathologies including macrovesicular steatosis and leads to disease progression. In this study, we measured the response to MCD diet in C57BL/6N mice obtained from three different sources; Korea NIFDS, USA, and Japan. We evaluated changes in body weight, food consumption, and relative weights of tissues such as liver, kidney, gonadal white adipose tissue, inguinal white adipose tissue, and brown adipose tissue. These basic parameters of mice with an MCD diet were not significantly different among the sources of mice tested. After 3 weeks on an MCD diet, histopathological analyses showed that the MCD diet induced clear fat vacuoles involving most area of the acinus in the liver of all mice. It was accompanied by increased serum activities of alanine aminotransferase and aspartate aminotransferase, and decreased levels of serum triglyceride and cholesterol. In conclusion, the response of C57BL6N mice originating from different sources to the MCD diet showed no significant differences as measured by physiological, biochemical, and histopathological parameters.

References

  1. 1.

    Loomba R, Sanyal AJ. The global NAFLD epidemic. Nat Rev Gastroenterol Hepatol 2013; 10(11): 686–690.

    CAS  Article  Google Scholar 

  2. 2.

    Chalasani N, Younossi Z, Lavine JE, Diehl AM, Brunt EM, Cusi K, Charlton M, Sanyal AJ; American Gastroenterological Association; American Association for the Study of Liver Diseases; American College of Gastroenterology!!. The diagnosis and management of non-alcoholic fatty liver disease: practice guideline by the American Gastroenterological Association, American Association for the Study of Liver Diseases, and American College of Gastroenterology. Gastroenterology 2012; 142(7): 1592–1609.

    Google Scholar 

  3. 3.

    Browning JD, Horton JD. Molecular mediators of hepatic steatosis and liver injury. J Clin Invest 2004; 114(2): 147–152.

    CAS  PubMed  Google Scholar 

  4. 4.

    Cohen JC, Horton JD, Hobbs HH. Human fatty liver disease: old questions and new insights. Science 2011; 332(6037): 1519–1523.

    CAS  PubMed  Google Scholar 

  5. 5.

    Day CP, James OF. Steatohepatitis: a tale of two “hits”? Gastroenterology 1998; 114(4): 842–845.

    CAS  PubMed  Google Scholar 

  6. 6.

    Kirsch R, Clarkson V, Shephard EG, Marais DA, Jaffer MA, Woodburne VE, Kirsch RE, Hall Pde L. Rodent nutritional model of non-alcoholic steatohepatitis: species, strain and sex difference studies. J Gastroenterol Hepatol 2003; 18(11): 1272–1282.

    PubMed  Google Scholar 

  7. 7.

    Leclercq IA, Farrell GC, Field J, Bell DR, Gonzalez FJ, Robertson GR. CYP2E1 and CYP4A as microsomal catalysts of lipid peroxides in murine nonalcoholic steatohepatitis. J Clin Invest 2000; 105(8): 1067–1075.

    CAS  PubMed  Google Scholar 

  8. 8.

    Bothe GW, Bolivar VJ, Vedder MJ, Geistfeld JG. Genetic and behavioral differences among five inbred mouse strains commonly used in the production of transgenic and knockout mice. Genes Brain Behav 2004; 3(3): 149–157.

    CAS  PubMed  Google Scholar 

  9. 9.

    Bryant CD, Zhang NN, Sokoloff G, Fanselow MS, Ennes HS, Palmer AA, McRoberts JA. Behavioral differences among C57BL/6 substrains: implications for transgenic and knockout studies. J Neurogenet 2008; 22(4): 315–331.

    CAS  PubMed  Google Scholar 

  10. 10.

    Mulligan MK, Ponomarev I, Boehm SL 2nd, Owen JA, Levin PS, Berman AE, Blednov YA, Crabbe JC, Williams RW, Miles MF, Bergeson SE. Alcohol trait and transcriptional genomic analysis of C57BL/6 substrains. Genes Brain Behav 2008; 7(6): 677–689.

    CAS  PubMed  Google Scholar 

  11. 11.

    Mekada K, Abe K, Murakami A, Nakamura S, Nakata H, Moriwaki K, Obata Y, Yoshiki A. Genetic differences among C57BL/6 substrains. Exp Anim 2009; 58(2): 141–149.

    CAS  PubMed  Google Scholar 

  12. 12.

    Zurita E, Chagoyen M, Cantero M, Alonso R, Gonzalez-Neira A, Lopez-Jimenez A, Lopez-Moreno JA, Landel CP, Benitez J, Pazos F, Montoliu L. Genetic polymorphisms among C57BL/6 mouse inbred strains. Transgenic Res 2011; 20(3): 481–489.

    CAS  PubMed  Google Scholar 

  13. 13.

    Itagaki H, Shimizu K, Morikawa S, Ogawa K, Ezaki T. Morphological and functional characterization of non-alcoholic fatty liver disease induced by a methionine-choline-deficient diet in C57BL/6 mice. Int J Clin Exp Pathol 2013; 6(12): 2683–2696.

    PubMed  Google Scholar 

  14. 14.

    Machado MV, Cortez-Pinto H. Non-alcoholic fatty liver disease: what the clinician needs to know. World J Gastroenterol 2014; 20(36): 12956–12980.

    CAS  PubMed  Google Scholar 

  15. 15.

    Rinella ME, Elias MS, Smolak RR, Fu T, Borensztajn J, Green RM. Mechanisms of hepatic steatosis in mice fed a lipogenic methionine choline-deficient diet. J Lipid Res 2008; 49(5): 1068–1076.

    CAS  PubMed  Google Scholar 

  16. 16.

    Yao ZM, Vance DE. The active synthesis of phosphatidylcholine is required for very low density lipoprotein secretion from rat hepatocytes. J Biol Chem 1988; 263(6): 2998–3004.

    CAS  PubMed  Google Scholar 

  17. 17.

    Caballero F, Fernandez A, Matias N, Martinez L, Fucho R, Elena M, Caballeria J, Morales A, Fernandez-Checa JC, Garcia-Ruiz C. Specific contribution of methionine and choline in nutritional nonalcoholic steatohepatitis: impact on mitochondrial S-adenosyl-L-methionine and glutathione. J Biol Chem 2010; 285(24): 18528–18536.

    CAS  PubMed  Google Scholar 

  18. 18.

    REITMAN S, FRANKEL S. A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clin Pathol 1957; 28(1): 56–63.

    CAS  PubMed  Google Scholar 

  19. 19.

    Wei E, Ben Ali Y, Lyon J, Wang H, Nelson R, Dolinsky VW, Dyck JR, Mitchell G, Korbutt GS, Lehner R. Loss of TGH/Ces3 in mice decreases blood lipids, improves glucose tolerance, and increases energy expenditure. Cell Metab 2010; 11(3): 183–193.

    CAS  PubMed  Google Scholar 

  20. 20.

    Kim SN, Jung YS, Kwon HJ, Seong JK, Granneman JG, Lee YH. Sex differences in sympathetic innervation and browning of white adipose tissue of mice. Biol Sex Differ 2016; 7: 67.

    PubMed  Google Scholar 

  21. 21.

    Tanaka N, Takahashi S, Fang ZZ, Matsubara T, Krausz KW, Qu A, Gonzalez FJ. Role of white adipose lipolysis in the development of NASH induced by methionine- and choline-deficient diet. Biochim Biophys Acta 2014; 1841(11): 1596–1607.

    CAS  PubMed  Google Scholar 

  22. 22.

    Day CP. Non-alcoholic fatty liver disease: a massive problem. Clin Med (Lond) 2011; 11(2): 176–178.

    Google Scholar 

  23. 23.

    Ekstedt M, Franzen LE, Mathiesen UL, Thorelius L, Holmqvist M, Bodemar G, Kechagias S. Long-term follow-up of patients with NAFLD and elevated liver enzymes. Hepatology 2006; 44(4): 865–873.

    CAS  PubMed  Google Scholar 

  24. 24.

    Sanyal AJ, Brunt EM, Kleiner DE, Kowdley KV, Chalasani N, Lavine JE, Ratziu V, McCullough A. Endpoints and clinical trial design for nonalcoholic steatohepatitis. Hepatology 2011; 54(1): 344–353.

    PubMed  Google Scholar 

  25. 25.

    Takahashi Y, Soejima Y, Fukusato T. Animal models of nonalcoholic fatty liver disease/nonalcoholic steatohepatitis. World J Gastroenterol 2012; 18(19): 2300–2308.

    PubMed  Google Scholar 

  26. 26.

    Sahai A, Malladi P, Melin-Aldana H, Green RM, Whitington PF. Upregulation of osteopontin expression is involved in the development of nonalcoholic steatohepatitis in a dietary murine model. Am J Physiol Gastrointest Liver Physiol 2004; 287(1): G264-273.

    Google Scholar 

  27. 27.

    Weltman MD, Farrell GC, Liddle C. Increased hepatocyte CYP2E1 expression in a rat nutritional model of hepatic steatosis with inflammation. Gastroenterology 1996; 111(6): 1645–1653.

    CAS  Google Scholar 

  28. 28.

    London RM, George J. Pathogenesis of NASH: animal models. Clin Liver Dis 2007; 11(1): 55–74, viii.

    PubMed  Google Scholar 

  29. 29.

    Yao ZM, Vance DE. Reduction in VLDL, but not HDL, in plasma of rats deficient in choline. Biochem Cell Biol 1990; 68(2): 552–558.

    CAS  Google Scholar 

  30. 30.

    Rinella ME, Green RM. The methionine-choline deficient dietary model of steatohepatitis does not exhibit insulin resistance. J Hepatol 2004; 40(1): 47–51.

    CAS  Google Scholar 

  31. 31.

    Machado MV, Michelotti GA, Xie G, Almeida Pereira T, Boursier J, Bohnic B, Guy CD, Diehl AM. Mouse models of diet-induced nonalcoholic steatohepatitis reproduce the heterogeneity of the human disease. PLoS One 2015; 10(5): e0127991.

    PubMed  Google Scholar 

  32. 32.

    Mashimo T, Serikawa T. Rat resources in biomedical research. Curr Pharm Biotechnol 2009; 10(2): 214–220.

    CAS  PubMed  Google Scholar 

  33. 33.

    Cray BA. The 1999 Reginald Thomson Lecture. Custom-built mice: unique discovery tools in biomedical research. Can Vet J 2000; 41(3): 201–206.

    Google Scholar 

  34. 34.

    Ardaillou R. Transgenic mice: a major advance in biomedical research. Bull Acad Natl Med 2009; 193(8): 1773–1782.

    CAS  Google Scholar 

  35. 35.

    Fontaine DA, Davis DB. Attention to Background Strain Is Essential for Metabolic Research: C57BL/6 and the International Knockout Mouse Consortium. Diabetes 2016; 65(1): 25–33.

    CAS  PubMed  Google Scholar 

  36. 36.

    Collins S, Martin TL, Surwit RS, Robidoux J. Genetic vulnerability to diet-induced obesity in the C57BL/6J mouse: physiological and molecular characteristics. Physiol Behav 2004; 81(2): 243–248.

    CAS  PubMed  Google Scholar 

  37. 37.

    Winzell MS, Ahren B. The high-fat diet-fed mouse: a model for studying mechanisms and treatment of impaired glucose tolerance and type 2 diabetes. Diabetes 2004; 53 Suppl 3: S215–219.

    Google Scholar 

  38. 38.

    Sundberg JP, Schofield PN. Commentary: mouse genetic nomenclature. Standardization of strain, gene, and protein symbols. Vet Pathol 2010; 47(6): 1100–1104.

    CAS  PubMed  Google Scholar 

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Acknowledgments

This project was supported by a grant of BIOREIN (Laboratory Animal Bio Resources Initiative) from Ministry of Food and Drug Safety in 2015.

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Correspondence to Young-Suk Jung.

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This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Kim, S.H., Lim, Y., Park, J.B. et al. Comparative study of fatty liver induced by methionine and choline-deficiency in C57BL/6N mice originating from three different sources. Lab Anim Res 33, 157–164 (2017). https://doi.org/10.5625/lar.2017.33.2.157

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Keywords

  • Non-alcoholic fatty liver disease
  • methionine-choline deficient diet
  • C57BL/6N