Journal of Gastroenterology

, Volume 48, Issue 5, pp 620–632 | Cite as

Deletion of Nrf2 leads to rapid progression of steatohepatitis in mice fed atherogenic plus high-fat diet

  • Kosuke Okada
  • Eiji Warabi
  • Hirokazu Sugimoto
  • Masaki Horie
  • Naohiro Gotoh
  • Katsutoshi Tokushige
  • Etsuko Hashimoto
  • Hirotoshi Utsunomiya
  • Hiroshi Takahashi
  • Tetsuro Ishii
  • Masayuki Yamamoto
  • Junichi ShodaEmail author
Original Article—Liver, Pancreas, and Biliary Tract



The transcription factor nuclear factor-E2-related factor-2 (Nrf2) inhibits lipid accumulation and oxidative stress in the liver by interfering with lipogenic pathways and inducing antioxidative stress genes.


The involvement of Nrf2 in defense against the development of steatohepatitis was studied in an experimental model induced by an atherogenic plus high-fat (Ath + HF) diet. Wild-type (WT) and Nrf2-null mice were fed the diet. Their specimens were analyzed for pathology as well as for the expression levels of genes involved in fatty acid metabolism and those involved via the Nrf2 transcriptional pathway.


In Nrf2-null mice fed the diet, steatohepatitis developed rapidly, leading to precirrhosis. The Ath + HF diet increased hepatic triglyceride levels and changed fatty acid composition in both mouse groups. However, oleic acid (C18:1 n-9) predominated in the livers of Nrf2-null mice. Correlating well with the pathology, the mRNA levels of the factors involved in fatty acid metabolism (Lxr, Srebp-1a, 1c, Acc-1, Fas, Scd-1, and Fatty acid transporting peptides 1, 3, 4), the inflammatory cytokine genes (Tnf-α and IL-), and the fibrogenesis-related genes (Tgf-β1 and α-Sma) were significantly increased in the livers of Nrf2-null mice fed the diet, compared with the levels of these factors in matched WT mice. Oxidative stress was significantly increased in the livers of Nrf2-null mice fed the diet. This change was closely associated with the decreased levels of antioxidative stress genes.


Nrf2 deletion leads to the rapid onset and progression of steatohepatitis induced by an Ath + HF diet, through both up-regulation of co-regulators of fatty acid metabolism and down-regulation of oxidative metabolism regulators in the liver.


Nrf2 gene-knockout mouse Transcription factor Atherogenic plus high-fat diet Fatty acid Oxidative stress 



Acetyl-CoA carboxylase


Acyl-CoA oxidase


Alpha-smooth muscle actin

Ath + HF diet

Atherogenic plus high-fat diet


Alanine aminotransferase


Aspartate aminotransferase


Carnitine palmitoyltransferase


Elongation of long-chain fatty acids family member 6


Fatty acid synthase


Fatty acid transport protein


γ-Glutamyl cystein synthetase




Glutathione S-transferase




Kelch-like Ech-associated protein 1


Liver fatty acid binding protein


Liver X receptor


Non-alcoholic steatohepatitis


Nuclear factor-E2-related factor-2


NAD(P)H: quinone oxidoreductase 1


Peroxisome proliferators activated receptor


Reactive oxygen species


Stearoyl-CoA desaturase-1


Sterol regulatory element-binding protein


Transforming growth factor


Wild type


Conflict of interest

The authors declare that they have no conflicts of interest.


  1. 1.
    Vuppalanchi R, Naga C. Nonalcoholic fatty liver disease and nonalcoholic steatohepatitis: selected practical issues in their evaluation and management. Hepatology. 2009;49:306–17.PubMedCrossRefGoogle Scholar
  2. 2.
    Torres DM, Harrison SA. Diagnosis and therapy of nonalcoholic steatohepatitis. Gastroenterology. 2008;134:1682–98.PubMedCrossRefGoogle Scholar
  3. 3.
    Marchesini G, Bugianesi E, Forlani G, Cerrelli F, Lenzi M, Manini R, et al. Nonalcoholic fatty liver, steatohepatitis, and metabolic syndrome. Hepatology. 2003;37:917–23.PubMedCrossRefGoogle Scholar
  4. 4.
    Browning JD, Horton JD. Molecular mediators of hepatic steatosis and liver injury. J Clin Invest. 2004;114:147–52.PubMedGoogle Scholar
  5. 5.
    Malaguarnera L, Madeddu R, Palio E, Arena N, Malaguarnera M. Heme oxygenase-1 levels and oxidative stress-related parameters in non-alcoholic fatty liver disease patients. J Hepatol. 2005;42:585–91.PubMedCrossRefGoogle Scholar
  6. 6.
    Sumida Y, Nakashima T, Yoh T, Furutani M, Hirohama A, Kakisaka Y, et al. Serum thioredoxin levels as a predictor of steatohepatitis in patients with nonalcoholic fatty liver disease. J Hepatol. 2003;38:32–8.PubMedCrossRefGoogle Scholar
  7. 7.
    Kotronen A, Seppala-Lindroos A, Bergholm R, Yki-Jarvinen H. Tissue specificity of insulin resistance in humans: fat in the liver rather than muscle is associated with features of the metabolic syndrome. Diabetologia. 2008;51:130–8.PubMedCrossRefGoogle Scholar
  8. 8.
    Fromenty B, Robin MA, Igoudili A, Mansouri A, Pessayre D. The ins and outs of mitochondrial dysfunction in NASH. Diabetes Metab 2004;30:121–38.Google Scholar
  9. 9.
    Yang S, Zhu H, Li Y, Lin H, Gabrielson K, Trush MA, et al. Mitochondrial adaptations to obesity related oxidant stress. Arch Biochem Biophys. 2000;378:259–68.PubMedCrossRefGoogle Scholar
  10. 10.
    Feldstein AE, Werneburg NW, Canbay A, Guicciardi ME, Bronk SF, Rydzewski R, et al. Free fatty acids promote hepatic lipotoxicity by stimulating TNF-α expression via a lysosomal pathway. Hepatology. 2004;40:185–94.PubMedCrossRefGoogle Scholar
  11. 11.
    Allard JP, Aghdassi E, Mohammed S, Raman M, Avand G, Arendt BM, et al. Nutritional assessment and hepatic fatty acid composition in non-alcoholic fatty liver disease (NAFLD): a cross-sectional study. J Hepatol. 2008;48:300–7.PubMedCrossRefGoogle Scholar
  12. 12.
    Zhang DD, Hannink M. Distinct cysteine in Keap1 are required for Keap1-dependent ubiquitination on Nrf2 and for stabilization of Nrf2 by chemopreventive agents and oxidative stress. Mol Cell Biol. 2003;23:8137–51.PubMedCrossRefGoogle Scholar
  13. 13.
    Kwak MK, Itoh K, Yamamoto M, Sutter TR, Kensler TW. Role of transcription factor Nrf2 in the induction of hepatic phase 2 and antioxidative enzymes in vivo by the cancer chemoprotective agent, 3H-1, 2-dithiole-3-thione. Mol Med. 2001;7:135–45.PubMedGoogle Scholar
  14. 14.
    Okada K, Shoda J, Taguchi K, Maher JM, Ishizaki K, Inoue Y, et al. Ursodeoxycholic acid stimulates Nrf2-mediated hepatocellular transport, detoxification, and antioxidative stress systems in mice. Am J Physiol Gastrointest Liver Pysiol. 2008;295:G735–47.CrossRefGoogle Scholar
  15. 15.
    Okada K, Shoda J, Taguchi K, Maher JM, Ishizaki K, Inoue Y, et al. Nrf2 counteracts cholestatic liver injury via stimulation of hepatic defense systems. Biochem Biophys Res Commun. 2009;389:431–6.PubMedCrossRefGoogle Scholar
  16. 16.
    Tanaka Y, Aleksunes LM, Yeager RL, Gyamfi MA, Esterly N, Guo GL, et al. NF-E2-related factor 2 inhibits lipid accumulation and oxidative stress in mice fed high-fat diet. J Pharmacol Exp Ther. 2008;325:655–64.PubMedCrossRefGoogle Scholar
  17. 17.
    Shin S, Wakabayashi J, Yates MS, Wakabayashi N, Dolan PM, Aja S, et al. Role of Nrf2 in prevention of high-fat diet-induced obesity by synthetic triterpenoid CDDO-Imidazolide. Eur J Pharmacol. 2009;620:138–44.PubMedCrossRefGoogle Scholar
  18. 18.
    Kitteringham N, Abdullah A, Walsh J, Randle L, Jenkins RE, Sison R, et al. Proteomic analysis of Nrf2 deficient transgenic mice reveals cellular defense and lipid metabolism as primary Nrf2-dependent pathways in the liver. J Proteomics. 2010;73:1612–31.PubMedCrossRefGoogle Scholar
  19. 19.
    Matsuzawa N, Takamura T, Kurita S, Nisu H, Ota T, Ando H, et al. Lipid-induced oxidative stress causes steatohepatitis in mice fed an atherogenic diet. Hepatology. 2007;46:1392–403.PubMedCrossRefGoogle Scholar
  20. 20.
    Sugimoto H, Okada K, Shoda J, Warabi E, Ishige K, Ueda T, et al. Deletion of nuclear factor-E2-related factor-2 leads to rapid onset and progression of nutritional steatohepatitis in mice. Am J Physiol Gastrointest Liver Physiol. 2010;298:G283–94.PubMedCrossRefGoogle Scholar
  21. 21.
    Gotoh N, Nagao K, Onoda S, Shirouchi B, Furuya K, Nagai T, et al. Effects of three different highly purified n-3 series highly unsaturated fatty acids on lipid metabolism in C57BL/KsJ-db/db mice. J Agric Food Chem. 2009;57:11047–54.PubMedCrossRefGoogle Scholar
  22. 22.
    Matsuzaka T, Shimano H, Yahagi N, Kato T, Atsumi A, Yamamoto T, et al. Crucial role of a long-chain fatty acid elongase, Elovl6, in obesity-induced insulin resistance. Nat Med. 2007;13:1193–202.PubMedCrossRefGoogle Scholar
  23. 23.
    Matsuzaka T, Shimano H. Elovl6: a new player in fatty acid metabolism and insulin sensitivity. J Mol Med. 2009;87:379–84.PubMedCrossRefGoogle Scholar
  24. 24.
    Maruyama A, Tsukamoto S, Nishikawa K, Yoshida A, Harada N, Motojima K, et al. Nrf2 regulates the alternative first exons of CD36 in macrophages through specific antioxidant response elements. Arch Biochem Biophys. 2008;477:139–45.PubMedCrossRefGoogle Scholar
  25. 25.
    Schwenk RW, Holloway GP, Luiken JJFP, Bonen A, Glatz JFC. Fatty acid transport across the cell membrane: regulation by fatty acid transporters. Prostaglandins Leukot Essent Fatty Acids. 2010;82:149–54.PubMedCrossRefGoogle Scholar
  26. 26.
    Tomita K, Oike Y, Teratani T, Taguchi T, Noguchi M, Suzuki T, et al. Hepatic adipoR2 signaling plays a protective role against progression of nonalcoholic steatohepatitis in mice. Hepatology. 2008;48:458–73.PubMedCrossRefGoogle Scholar
  27. 27.
    Adams LA, Lymp JF, Sauver JS, Sanderson SO, Lindor KD, Feldstein A, et al. The natural history of nonalcoholic fatty liver disease: a population based cohort study. Gastroenterology. 2005;129:113–21.PubMedCrossRefGoogle Scholar
  28. 28.
    Puri P, Wiest MM, Cheung O, Mirshahi F, Sargeant C, Min HK, et al. The plasma lipidomic signature of nonalcoholic steatohepatitis. Hepatology. 2009;50:1827–38.PubMedCrossRefGoogle Scholar
  29. 29.
    Araya J, Rodrigo R, Vidla LA, Thielemann L, Orellana M, Pettinelli P, et al. Increase in long-chain polyunsaturated fatty acid n-6/n-3 ratio in relation to hepatic steatosis in patients with non-alcoholic fatty liver disease. Clin Sci. 2004;106:635–43.PubMedCrossRefGoogle Scholar
  30. 30.
    Moriya K, Todoroki T, Tsutsumi T, Fujie H, Shintani Y, Miyoshi H, et al. Increase in the concentration of carbon 18 monounsaturated fatty acids in the liver with hepatitis C: analysis in transgenic mice and humans. Biochem Biophys Res Commun. 2001;281:1207–12.PubMedCrossRefGoogle Scholar
  31. 31.
    Feldstein A, Canbay A, Guicciardi ME, Higuchi H, Bronk SF, Gores GJ. Diet associated hepatic steatosis sensitizes to Fas mediated liver injury in mice. J Hepatol. 2003;39:978–83.PubMedCrossRefGoogle Scholar
  32. 32.
    Mao J, DeMayo FJ, Li H, Abu-Elheiga L, Gu Z, Shaikenov TE, et al. Liver-specific deletion of acetyl-CoA carboxylase 1 reduces hepatic triglyceride accumulation without affecting glucose homeostasis. Proc Natl Acad Sci USA. 2006;103:8552–7.PubMedCrossRefGoogle Scholar
  33. 33.
    Ntambi JM, Miyazaki M, Stoehr JP, Lan H, Kendziorski CM, Yandell BS, et al. Loss of stearoyl-CoA desaturase-1 function protects mice against adiposity. Proc Natl Acad Sci USA. 2002;99:11482–6.PubMedCrossRefGoogle Scholar
  34. 34.
    Miyazaki M, Flowers MT, Sampath H, Chu K, Otzelberger C, Liu X, et al. Hepatic stearoyl-CoA desaturase-1 deficiency protects mice from carbohydrate-induced adiposity and hepatic steatosis. Cell Metab. 2007;6:484–96.PubMedCrossRefGoogle Scholar
  35. 35.
    Miquilena-Colina ME, Lima-Cabello E, Sanchez-Campos S, Garcia-Mediavilla MV, Fernandez-Beremejo M, Lazano-Rodriguez T, et al. Hepatic fatty acid translocase CD36 upregulation is associated with insulin resistance, hyperinsulinaemia and increased steatosis in non-alcoholic steatohepatitis and chronic hepatitis C. Gut. 2011;60:1394–402.PubMedCrossRefGoogle Scholar
  36. 36.
    Pi J, Leung L, Xue P, Wang W, Hou Y, Liu D, et al. Deficiency in the nuclear factor E2-related factor-2 transcription factor results in impaired adipogenesis and protects against diet-induced obesity. J Biol Chem. 2010;285:9292–300.PubMedCrossRefGoogle Scholar
  37. 37.
    Chartoumpekis DV, Ziros PG, Psyrogiannis AI, Papavassilou AG, Kyriazopoulou VE, Sykiotis GP, et al. Nrf2 represses FGF21 during long-term high-fat diet-induced obesity in mice. Diabetes. 2011;60:2465–73.PubMedCrossRefGoogle Scholar

Copyright information

© Springer 2012

Authors and Affiliations

  • Kosuke Okada
    • 1
    • 2
  • Eiji Warabi
    • 3
  • Hirokazu Sugimoto
    • 2
  • Masaki Horie
    • 1
  • Naohiro Gotoh
    • 4
  • Katsutoshi Tokushige
    • 5
  • Etsuko Hashimoto
    • 5
  • Hirotoshi Utsunomiya
    • 6
  • Hiroshi Takahashi
    • 7
  • Tetsuro Ishii
    • 3
  • Masayuki Yamamoto
    • 8
  • Junichi Shoda
    • 1
    Email author
  1. 1.Division of Medical Science, Faculty of MedicineUniversity of TsukubaTsukubaJapan
  2. 2.Department of Gastroenterology, Faculty of MedicineThe University of TsukubaTsukubaJapan
  3. 3.Division of Biomedical Sciences, Faculty of MedicineUniversity of TsukubaTsukubaJapan
  4. 4.Department of Food Science and TechnologyTokyo University of Marine Science and TechnologyTokyoJapan
  5. 5.Department of Internal Medicine and GastroenterologyTokyo Women’s Medical UniversityTokyoJapan
  6. 6.Department of Strategic Surveillance for Functional Food and Comprehensive Traditional MedicineWakayama Medical UniversityWakayamaJapan
  7. 7.Department of Anesthesiology, Faculty of MedicineUniversity of TsukubaTsukubaJapan
  8. 8.Department of Medical BiochemistryTohoku University Graduate School of MedicineSendaiJapan

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