Advertisement

Molecular Medicine

, Volume 19, Issue 1, pp 237–244 | Cite as

Nrf2 Activates Augmenter of Liver Regeneration (ALR) via Antioxidant Response Element and Links Oxidative Stress to Liver Regeneration

  • Rania Dayoub
  • Arndt Vogel
  • Jutta Schuett
  • Madeleine Lupke
  • Susannah M. Spieker
  • Nadja Kettern
  • Eberhard Hildt
  • Michael Melter
  • Thomas S. Weiss
Research Article

Abstract

Liver regeneration can be impaired by permanent oxidative stress and activation of nuclear factor erythroid 2-related factor 2 (Nrf2), known to regulate the cellular antioxidant response, and has been shown to improve the process of liver regeneration. A variety of factors regulate hepatic tissue regeneration, among them augmenter of liver regeneration (ALR), attained great attention as being survival factors for the liver with proproliferative and antiapoptotic properties. Here we determined the Nrf2/ antioxidant response element (ARE) regulated expression of ALR and show ALR as a target gene of Nrf2 in vitro and in vivo. The ALR promoter comprises an ARE binding site and, therefore, ALR expression can be induced by ARE-activator tertiary butylhydroquinone (tBHQ) in hepatoma cells and primary human hepatocytes (PHH). Promoter activity and expression of ALR were enhanced after cotransfection of Nrf2 compared with control and dominant negative mutant of Nrf2. Performing partial hepatectomy in livers from Nrf2+/+ mice compared with Nrf2−/− knock-out (KO) mice, we found increased expression of ALR in addition to known antioxidant ARE-regulated genes. Furthermore, we observed increased ALR expression in hepatitis B virus (HBV) compared with hepatitis C virus (HCV) positive hepatoma cells and PHH. Recently, it was demonstrated that HBV infection activates Nrf2 and, now, we add results showing increased ALR expression in liver samples from patients infected with HBV. ALR is regulated by Nrf2, acts as a liver regeneration and antioxidative protein and, therefore, links oxidative stress to hepatic regeneration to ensure survival of damaged cells.

Notes

Acknowledgments

The authors are grateful to Friederike Schrenk, Elke Gerstl and Renate Lange, Department of Pediatrics and Juvenile Medicine, University of Regensburg Hospital, Germany, for their excellent technical assistance. This work was supported by the Excellence Cluster REBIRTH for A Vogel, the Medical Faculty of the University of Regensburg ReForM-C for TS Weiss and the German Federal Ministry of Education and Research (BMBF) Virtual Liver Network grant FKZ 0315753 to TS Weiss.

References

  1. 1.
    Fausto N. (2000) Liver regeneration. J. Hepatol. 32 Suppl 1:19–31.CrossRefPubMedGoogle Scholar
  2. 2.
    Schwabe RF, Brenner DA. (2006) Mechanisms of liver injury. I. TNF-alpha-induced liver injury: role of IKK, JNK, and ROS pathways. Am. J. Physiol. Gastrointest. Liver Physiol. 290:G583–9.CrossRefPubMedGoogle Scholar
  3. 3.
    Wasserman WW, Fahl WE. (1997) Functional antioxidant responsive elements. Proc. Natl. Acad. Sci. U. S. A. 94:5361–6.CrossRefGoogle Scholar
  4. 4.
    Kobayashi M, Yamamoto M. (2005) Molecular mechanisms activating the Nrf2-Keap1 pathway of antioxidant gene regulation. Antioxid. Redox. Signal. 7:385–94.CrossRefPubMedGoogle Scholar
  5. 5.
    Jaiswal AK. (2004) Nrf2 signaling in coordinated activation of antioxidant gene expression. Free Radic. Biol. Med. 36:1199–207.CrossRefPubMedGoogle Scholar
  6. 6.
    Nguyen T, Sherratt PJ, Pickett CB. (2003) Regulatory mechanisms controlling gene expression mediated by the antioxidant response element. Annu. Rev. Pharmacol. Toxicol. 43:233–60.CrossRefPubMedGoogle Scholar
  7. 7.
    Lee JM, et al. (2005) Nrf2, a multi-organ protector? FASEB J. 19:1061–6.CrossRefPubMedGoogle Scholar
  8. 8.
    Beyer TA, et al. (2008) Impaired liver regeneration in Nrf2 knockout mice: role of ROS-mediated insulin/IGF-1 resistance. EMBO J. 27:212–23.CrossRefPubMedGoogle Scholar
  9. 9.
    Xu W, et al. (2008) The Nrf2 transcription factor protects from toxin-induced liver injury and fibrosis. Lab. Invest. 88:1068–78.CrossRefPubMedGoogle Scholar
  10. 10.
    Schaedler S, et al. (2010) Hepatitis B virus induces expression of antioxidant response elementregulated genes by activation of Nrf2. J. Biol. Chem. 285:41074–86.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Carvajal-Yepes M, et al. (2011) Hepatitis C virus impairs the induction of cytoprotective Nrf2 target genes by delocalization of small Maf proteins. J. Biol. Chem. 286:8941–51.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Pawlowski R, Jura J. (2006) ALR and liver regeneration. Mol. Cell Biochem. 288:159–69.CrossRefPubMedGoogle Scholar
  13. 13.
    Polimeno L, et al. (2011) Alrp, a survival factor that controls the apoptotic process of regenerating liver after partial hepatectomy in rats. Free Radic. Res. 45:534–49.CrossRefPubMedGoogle Scholar
  14. 14.
    Zhang LM, et al. (2005) Effect of naked eukaryotic expression plasmid encoding rat augmenter of liver regeneration on acute hepatic injury and hepatic failure in rats. World J. Gastroenterol. 11:3680–5.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Margeli AP, et al. (2003) Hepatic stimulator substance administration ameliorates liver regeneration in an animal model of fulminant hepatic failure and encephalopathy. Liver Int. 23:171–8.CrossRefPubMedGoogle Scholar
  16. 16.
    Li Q, et al. (2005) Effects of augmentation of liver regeneration recombinant plasmid on rat hepatic fibrosis. World J. Gastroenterol. 11:2438–43.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Gatzidou E, Kouraklis G, Theocharis S. (2006) Insights on augmenter of liver regeneration cloning and function. World J. Gastroenterol. 12:4951–8.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Bihlmaier K, Mesecke N, Kloeppel C, Herrmann JM. (2008) The disulfide relay of the intermembrane space of mitochondria: an oxygen-sensing system? Ann. N. Y. Acad. Sci. 1147:293–302.CrossRefPubMedGoogle Scholar
  19. 19.
    Francavilla A, et al. (1994) Augmenter of liver regeneration: its place in the universe of hepatic growth factors. Hepatology. 20:747–57.CrossRefPubMedGoogle Scholar
  20. 20.
    Gandhi CR. (2012) Augmenter of liver regeneration. Fibrogenesis Tissue Repair. 5:10.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Li Y, et al. (2000) Stimulation of the mitogen-activated protein kinase cascade and tyrosine phosphorylation of the epidermal growth factor receptor by hepatopoietin. J. Biol. Chem. 275:37443–7.CrossRefPubMedGoogle Scholar
  22. 22.
    Ilowski M, et al. (2010) Augmenter of liver regeneration causes different kinetics of ERK1/2 and Akt/PKB phosphorylation than EGF and induces hepatocyte proliferation in an EGF receptor independent and liver specific manner. Biochem. Biophys. Res. Commun. 394:915–20.CrossRefPubMedGoogle Scholar
  23. 23.
    Polimeno L, et al. (2009) Protective effect of augmenter of liver regeneration on hydrogen peroxide-induced apoptosis in SH-SY5Y human neuroblastoma cells. Free Radic. Res. 43:865–75.CrossRefPubMedGoogle Scholar
  24. 24.
    Cao Y, et al. (2009) Human augmenter of liver regeneration is important for hepatoma cell viability and resistance to radiation-induced oxidative stress. Free Radic. Biol. Med. 47:1057–66.CrossRefPubMedGoogle Scholar
  25. 25.
    Wu Y, Chen L, Yu H, Liu H, An W. (2007) Transfection of hepatic stimulator substance gene desensitizes hepatoma cells to H2O2-induced cell apoptosis via preservation of mitochondria. Arch. Biochem. Biophys. 464:48–56.CrossRefPubMedGoogle Scholar
  26. 26.
    Thirunavukkarasu C, et al. (2008) Augmenter of liver regeneration: an important intracellular survival factor for hepatocytes. J. Hepatol. 48:578–88.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Todd LR, Gomathinayagam R, Sankar U. (2010) A novel Gfer-Drp1 link in preserving mitochondrial dynamics and function in pluripotent stem cells. Autophagy. 6:821–2.CrossRefPubMedGoogle Scholar
  28. 28.
    Hu R, et al. (2006) Gene expression profiles induced by cancer chemopreventive isothiocyanate sulforaphane in the liver of C57BL/6J mice and C57BL/6J/Nrf2 (−/−) mice. Cancer Lett. 243:170–92.CrossRefPubMedGoogle Scholar
  29. 29.
    Hu R, et al. (2006) Identification of Nrf2-regulated genes induced by chemopreventive isothiocyanate PEITC by oligonucleotide microarray. Life Sci. 79:1944–55.CrossRefPubMedGoogle Scholar
  30. 30.
    World Medical Association [WMA]. (1949) WMA international code of medical ethics. Last amended 2006 Oct. [cited 2013 Jul 25]. Available from: https://doi.org/www.wma.net/en/30publications/10policies/c8/
  31. 31.
    Weiss TS, Pahernik S, Scheruebl I, Jauch KW, Thasler WE. (2003) Cellular damage to human hepatocytes through repeated application of 5-aminolevulinic acid. J. Hepatol. 38:476–82.CrossRefGoogle Scholar
  32. 32.
    Dayoub R, et al. (2010) Foxa2 (HNF-3beta) regulates expression of hepatotrophic factor ALR in liver cells. Biochem. Biophys. Res. Commun. 395:465–70.CrossRefPubMedGoogle Scholar
  33. 33.
    Itoh K, et al. (1997) An Nrf2/small Maf heterodimer mediates the induction of phase II detoxifying enzyme genes through antioxidant response elements. Biochem. Biophys. Res. Commun. 236:313–22.CrossRefPubMedGoogle Scholar
  34. 34.
    Mitchell C, Willenbring H. (2008) A reproducible and well-tolerated method for 2/3 partial hepatectomy in mice. Nat. Protoc. 3:1167–70.CrossRefPubMedGoogle Scholar
  35. 35.
    Guidotti LG, Matzke B, Schaller H, Chisari FV. (1995) High-level hepatitis B virus replication in transgenic mice. J. Virol. 69:6158–69.PubMedPubMedCentralGoogle Scholar
  36. 36.
    Wakita T, et al. (2005) Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nat. Med. 11:791–6.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Himmelsbach K, et al. (2009) New aspects of an anti-tumour drug: sorafenib efficiently inhibits HCV replication. Gut. 58:1644–53.CrossRefPubMedGoogle Scholar
  38. 38.
    Lupberger J, Mund A, Kock J, Hildt E. (2006) Cultivation of HepG2.2.15 on Cytodex-3: higher yield of hepatitis B virus and less subviral particles compared to conventional culture methods. J. Hepatol. 45:547–52.CrossRefPubMedGoogle Scholar
  39. 39.
    Dayoub R, et al. (2011) Liver regeneration associated protein (ALR) exhibits antimetastatic potential in hepatocellular carcinoma. Mol. Med. 17:221–8.CrossRefPubMedGoogle Scholar
  40. 40.
    Liao XH, et al. (2010) Augmenter of liver regeneration protects kidneys from ischaemia/reperfusion injury in rats. Nephrol. Dial. Transplant. 25:2921–9.CrossRefPubMedGoogle Scholar
  41. 41.
    Zhao Y, et al. (2003) An initiator and its flanking elements function as a core promoter driving transcription of the Hepatopoietin gene. FEBS Lett 540:58–64.CrossRefPubMedGoogle Scholar
  42. 42.
    Dong LY, et al. (2007) Identification of human hepatic stimulator substance gene promoter and demonstration of dual regulation of AP1/AP4 cisacting element in different cell lines. Int. J. Biochem. Cell Biol. 39:181–96.CrossRefPubMedGoogle Scholar
  43. 43.
    Guo D, Dong LY, Wu Y, Yang L, An W. (2008) Down-regulation of hepatic nuclear factor 4alpha on expression of human hepatic stimulator substance via its action on the proximal promoter in HepG2 cells. Biochem. J. 415:111–21.CrossRefPubMedGoogle Scholar
  44. 44.
    Arai M, et al. (2003) Gene expression profiling reveals the mechanism and pathophysiology of mouse liver regeneration. J. Biol. Chem. 278:29813–8.CrossRefPubMedGoogle Scholar
  45. 45.
    Xu L, et al. (2001) Expression profiling suggested a regulatory role of liver-enriched transcription factors in human hepatocellular carcinoma. Cancer Res. 61:3176–81.PubMedGoogle Scholar
  46. 46.
    Malhi H, Gores GJ, Lemasters JJ. (2006) Apoptosis and necrosis in the liver: a tale of two deaths? Hepatology. 43:S31–S44.CrossRefPubMedGoogle Scholar
  47. 47.
    Lau A, Villeneuve NF, Sun Z, Wong PK, Zhang DD. (2008) Dual roles of Nrf2 in cancer. Pharmacol. Res. 58:262–70.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Thasler WE, et al. (2005) Expression of augmenter of liver regeneration (ALR) in human liver cirrhosis and carcinoma. Histopathology. 47:57–66.CrossRefPubMedGoogle Scholar
  49. 49.
    Lamle J, et al. (2008) Nuclear factor-eythroid 2-related factor 2 prevents alcohol-induced fulminant liver injury. Gastroenterology. 134:1159–68.CrossRefPubMedGoogle Scholar
  50. 50.
    Tan KP, Yang M, Ito S. (2007) Activation of nuclear factor (erythroid-2 like) factor 2 by toxic bile acids provokes adaptive defense responses to enhance cell survival at the emergence of oxidative stress. Mol. Pharmacol. 72:1380–90.CrossRefPubMedGoogle Scholar

Copyright information

© The Author(s) 2013

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (https://doi.org/creativecommons.org/licenses/by-nc-nd/4.0/)

Authors and Affiliations

  • Rania Dayoub
    • 1
    • 2
  • Arndt Vogel
    • 3
  • Jutta Schuett
    • 3
  • Madeleine Lupke
    • 1
  • Susannah M. Spieker
    • 1
  • Nadja Kettern
    • 4
  • Eberhard Hildt
    • 4
  • Michael Melter
    • 1
  • Thomas S. Weiss
    • 1
    • 2
  1. 1.Department of Pediatrics and Juvenile MedicineUniversity of Regensburg HospitalRegensburgGermany
  2. 2.Center for Liver Cell ResearchUniversity of Regensburg HospitalRegensburgGermany
  3. 3.Department of GastroenterologyHepatology and Endocrinology, Hannover Medical SchoolHannoverGermany
  4. 4.Department of VirologyPaul-Ehrlich-InstitutLangenGermany

Personalised recommendations