Skip to main content
Log in

Simvastatin activates Keap1/Nrf2 signaling in rat liver

  • Original Article
  • Published:
Journal of Molecular Medicine Aims and scope Submit manuscript

Abstract

Some of the statins’ pleiotropic actions have been attributed to their antioxidant activity. The Nrf2 transcription factor controls the expression of a number of protective genes in response to oxidative stress. In the present study, wistar rats, primary hepatocytes as well as ST2 cells, were employed to explore the potential role of Nrf2 in mediating the reported antioxidant effects of statins. Simvastatin triggered nuclear translocation of Nrf2 in rat liver and in primary rat hepatocytes in a mevalonate-dependent and cholesterol-independent way. In liver, nuclear extracts from simvastatin-treated rats, the DNA-binding activity of Nrf2, was significantly increased and the mRNA of two known targets of Nrf2 (HO-1 and GPX2) was induced. In ST2 cells stably transfected with constructs bearing Nrf2-binding site (antioxidant responsive element), simvastatin enhanced Nrf2-mediated transcriptional activity in a mevalonate-dependent and cholesterol-independent fashion. In conclusion, activation of Keap1/Nrf2 signaling pathway by simvastatin might provide effective protection of the cell from the deleterious effects of oxidative stress.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

ARE:

Antioxidant responsive element

DLS:

Delipidated serum

DMEM:

Dulbecco’s modified Eagle’s medium

DN:

Dominant negative

EMSA:

Electrophoretic mobility-shift assay

FBS:

Fetal bovine serum

GPX:

Glutathione peroxidase

HMG-CoA:

3-hydroxy-3-methylglutaryl coenzyme A

HO-1:

Heme oxygenase 1

NQO1:

NAD(P)H:quinone oxidoreductase 1

Nrf2:

NF-E2-related factor 2

ROS:

Reactive oxygen species

RPA:

Ribonuclease protection assay

SREBP-2:

Sterol regulatory element-binding protein 2

TRXR:

Thioredoxin reductase

References

  1. Liao JK, Laufs U (2005) Pleiotropic effects of statins. Annu Rev Pharmacol Toxicol 45:89–118

    Article  PubMed  CAS  Google Scholar 

  2. Buhaescu I, Izzedine H (2007) Mevalonate pathway: a review of clinical and therapeutical implications. Clin Biochem 40:575–584

    Article  PubMed  CAS  Google Scholar 

  3. Perez-Guerrero C, Alvarez de Sotomayor M, Jimenez L, Herrera MD, Marhuenda E (2003) Effects of simvastatin on endothelial function after chronic inhibition of nitric oxide synthase by L-NAME. J Cardiovasc Pharmacol 42:204–210

    Article  PubMed  CAS  Google Scholar 

  4. Carneado J, Alvarez de Sotomayor M, Perez-Guerrero C, Jimenez L, Herrera MD, Pamies E, Martin-Sanz MD, Stiefel P, Miranda M, Bravo L, Marhuenda E (2002) Simvastatin improves endothelial function in spontaneously hypertensive rats through a superoxide dismutase mediated antioxidant effect. J Hypertens 20:429–437

    Article  PubMed  CAS  Google Scholar 

  5. Hsu M, Muchova L, Morioka I, Wong RJ, Schroder H, Stevenson DK (2006) Tissue-specific effects of statins on the expression of heme oxygenase-1 in vivo. Biochem Biophys Res Commun 343:738–744

    Article  PubMed  CAS  Google Scholar 

  6. Moosmann B, Behl C (2004) Selenoprotein synthesis and side-effects of statins. Lancet 363:892–894

    Article  PubMed  CAS  Google Scholar 

  7. Suzuki T, Kelly VP, Motohashi H, Nakajima O, Takahashi S, Nishimura S, Yamamoto M (2008) Deletion of the selenocysteine tRNA gene in macrophages and liver results in compensatory gene induction of cytoprotective enzymes by Nrf2. J Biol Chem 283:2021–2030

    Article  PubMed  CAS  Google Scholar 

  8. Kobayashi M, Yamamoto M (2006) Nrf2–Keap1 regulation of cellular defense mechanisms against electrophiles and reactive oxygen species. Adv Enzyme Regul 46:113–140

    Article  PubMed  CAS  Google Scholar 

  9. Kensler TW, Wakabayashi N, Biswal S (2007) Cell survival responses to environmental stresses via the Keap1–Nrf2–ARE pathway. Annu Rev Pharmacol Toxicol 47:89–116

    Article  PubMed  CAS  Google Scholar 

  10. Dinkova-Kostova AT, Talalay P (2008) Direct and indirect antioxidant properties of inducers of cytoprotective proteins. Mol Nutr Food Res 52:128–138

    Google Scholar 

  11. Seglen PO (1976) Preparation of isolated rat liver cells. Methods Cell Biol 13:29–83

    Article  PubMed  CAS  Google Scholar 

  12. Habeos I, Ziros PG, Psyrogiannis A, Vagenakis AG, Papavassiliou AG (2005) Statins and transcriptional regulation: the FXR connection. Biochem Biophys Res Commun 334:601–605

    Article  PubMed  CAS  Google Scholar 

  13. Alam J, Stewart D, Touchard C, Boinapally S, Choi AM, Cook JL (1999) Nrf2, a Cap‘n’Collar transcription factor, regulates induction of the heme oxygenase-1 gene. J Biol Chem 274:26071–26078

    Article  PubMed  CAS  Google Scholar 

  14. Chen JC, Huang KC, Lin WW (2006) HMG-CoA reductase inhibitors upregulate heme oxygenase-1 expression in murine RAW264.7 macrophages via ERK, p38 MAPK and protein kinase G pathways. Cell Signal 18:32–39

    Article  PubMed  CAS  Google Scholar 

  15. Hsieh CH, Jeng SF, Hsieh MW, Chen YC, Rau CS, Lu TH, Chen SS (2008) Statin-induced heme oxygenase-1 increases NF-kappaB activation and oxygen radical production in cultured neuronal cells exposed to lipopolysaccharide. Toxicol Sci 102:150–159

    Article  PubMed  CAS  Google Scholar 

  16. Hsieh CH, Rau CS, Hsieh MW, Chen YC, Jeng SF, Lu TH, Chen SS (2008) Simvastatin-induced heme oxygenase-1 increases apoptosis of Neuro 2A cells in response to glucose deprivation. Toxicol Sci 101:112–121

    Article  PubMed  CAS  Google Scholar 

  17. Lee TS, Chang CC, Zhu Y, Shyy JY (2004) Simvastatin induces heme oxygenase-1: a novel mechanism of vessel protection. Circulation 110:1296–1302

    Article  PubMed  CAS  Google Scholar 

  18. Grosser N, Hemmerle A, Berndt G, Erdmann K, Hinkelmann U, Schurger S, Wijayanti N, Immenschuh S, Schroder H (2004) The antioxidant defense protein heme oxygenase 1 is a novel target for statins in endothelial cells. Free Radic Biol Med 37:2064–2071

    Article  PubMed  CAS  Google Scholar 

  19. Lai IR, Chang KJ, Tsai HW, Chen CF (2008) Pharmacological preconditioning with simvastatin protects liver from ischemia-reperfusion injury by heme oxygenase-1 induction. Transplantation 85:732–738

    PubMed  CAS  Google Scholar 

  20. Burk RF, Hill KE, Nakayama A, Mostert V, Levander XA, Motley AK, Johnson DA, Johnson JA, Freeman ML, Austin LM (2008) Selenium deficiency activates mouse liver Nrf2-ARE but vitamin E deficiency does not. Free Radic Biol Med 44:1617–1623

    Article  PubMed  CAS  Google Scholar 

  21. Wierzbicki AS, Poston R, Ferro A (2003) The lipid and non-lipid effects of statins. Pharmacol Ther 99:95–112

    Article  PubMed  CAS  Google Scholar 

  22. Davignon J, Jacob RF, Mason RP (2004) The antioxidant effects of statins. Coron Artery Dis 15:251–258

    Article  PubMed  Google Scholar 

  23. Stoll LL, McCormick ML, Denning GM, Weintraub NL (2004) Antioxidant effects of statins. Drugs Today (Barc) 40:975–990

    Article  CAS  Google Scholar 

  24. Beltowski J, Wojcicka G, Jamroz A (2004) Effect of 3-hydroxy-3-methylglutarylcoenzyme A reductase inhibitors (statins) on tissue paraoxonase 1 and plasma platelet activating factor acetylhydrolase activities. J Cardiovasc Pharmacol 43:121–127

    Article  PubMed  CAS  Google Scholar 

  25. Ali F, Hamdulay SS, Kinderlerer AR, Boyle JJ, Lidington EA, Yamaguchi T, Soares MP, Haskard DO, Randi AM, Mason JC (2007) Statin-mediated cytoprotection of human vascular endothelial cells: a role for Kruppel-like factor 2-dependent induction of heme oxygenase-1. J Thromb Haemost 5:2537–2546

    Article  PubMed  CAS  Google Scholar 

  26. McFarlane SI, Muniyappa R, Francisco R, Sowers JR (2002) Clinical review 145: pleiotropic effects of statins: lipid reduction and beyond. J Clin Endocrinol Metab 87:1451–1458

    Article  PubMed  CAS  Google Scholar 

  27. Moosmann B, Behl C (2004) Selenoproteins, cholesterol-lowering drugs, and the consequences: revisiting of the mevalonate pathway. Trends Cardiovasc Med 14:273–281

    Article  PubMed  CAS  Google Scholar 

  28. Weiss SL, Sunde RA (1998) Cis-acting elements are required for selenium regulation of glutathione peroxidase-1 mRNA levels. RNA 4:816–827

    Article  PubMed  CAS  Google Scholar 

  29. Wingler K, Bocher M, Flohe L, Kollmus H, Brigelius-Flohe R (1999) mRNA stability and selenocysteine insertion sequence efficiency rank gastrointestinal glutathione peroxidase high in the hierarchy of selenoproteins. Eur J Biochem 259:149–157

    Article  PubMed  CAS  Google Scholar 

  30. Katz MS (2005) Therapy insight: potential of statins for cancer chemoprevention and therapy. Nat Clin Pract Oncol 2:82–89

    Article  PubMed  CAS  Google Scholar 

  31. Tsui TY, Lau CK, Ma J, Wu X, Wang YQ, Farkas S, Xu R, Schlitt HJ, Fan ST (2005) rAAV-mediated stable expression of heme oxygenase-1 in stellate cells: a new approach to attenuate liver fibrosis in rats. Hepatology 42:335–342

    Article  PubMed  CAS  Google Scholar 

  32. Rombouts K, Kisanga E, Hellemans K, Wielant A, Schuppan D, Geerts A (2003) Effect of HMG-CoA reductase inhibitors on proliferation and protein synthesis by rat hepatic stellate cells. J Hepatol 38:564–572

    Article  PubMed  CAS  Google Scholar 

  33. Brigelius-Flohe R (2006) Glutathione peroxidases and redox-regulated transcription factors. Biol Chem 387:1329–1335

    Article  PubMed  CAS  Google Scholar 

  34. Banning A, Deubel S, Kluth D, Zhou Z, Brigelius-Flohe R (2005) The GI-GPx gene is a target for Nrf2. Mol Cell Biol 25:4914–4923

    Article  PubMed  CAS  Google Scholar 

Download references

Competing interests

The authors declare no competing interests.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Athanasios G. Papavassiliou.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 499 KB)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Habeos, I.G., Ziros, P.G., Chartoumpekis, D. et al. Simvastatin activates Keap1/Nrf2 signaling in rat liver. J Mol Med 86, 1279–1285 (2008). https://doi.org/10.1007/s00109-008-0393-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00109-008-0393-4

Keywords

Navigation