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.
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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
Liao JK, Laufs U (2005) Pleiotropic effects of statins. Annu Rev Pharmacol Toxicol 45:89–118
Buhaescu I, Izzedine H (2007) Mevalonate pathway: a review of clinical and therapeutical implications. Clin Biochem 40:575–584
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
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
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
Moosmann B, Behl C (2004) Selenoprotein synthesis and side-effects of statins. Lancet 363:892–894
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
Kobayashi M, Yamamoto M (2006) Nrf2–Keap1 regulation of cellular defense mechanisms against electrophiles and reactive oxygen species. Adv Enzyme Regul 46:113–140
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
Dinkova-Kostova AT, Talalay P (2008) Direct and indirect antioxidant properties of inducers of cytoprotective proteins. Mol Nutr Food Res 52:128–138
Seglen PO (1976) Preparation of isolated rat liver cells. Methods Cell Biol 13:29–83
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
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
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
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
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
Lee TS, Chang CC, Zhu Y, Shyy JY (2004) Simvastatin induces heme oxygenase-1: a novel mechanism of vessel protection. Circulation 110:1296–1302
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
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
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
Wierzbicki AS, Poston R, Ferro A (2003) The lipid and non-lipid effects of statins. Pharmacol Ther 99:95–112
Davignon J, Jacob RF, Mason RP (2004) The antioxidant effects of statins. Coron Artery Dis 15:251–258
Stoll LL, McCormick ML, Denning GM, Weintraub NL (2004) Antioxidant effects of statins. Drugs Today (Barc) 40:975–990
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
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
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
Moosmann B, Behl C (2004) Selenoproteins, cholesterol-lowering drugs, and the consequences: revisiting of the mevalonate pathway. Trends Cardiovasc Med 14:273–281
Weiss SL, Sunde RA (1998) Cis-acting elements are required for selenium regulation of glutathione peroxidase-1 mRNA levels. RNA 4:816–827
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
Katz MS (2005) Therapy insight: potential of statins for cancer chemoprevention and therapy. Nat Clin Pract Oncol 2:82–89
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
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
Brigelius-Flohe R (2006) Glutathione peroxidases and redox-regulated transcription factors. Biol Chem 387:1329–1335
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
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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
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DOI: https://doi.org/10.1007/s00109-008-0393-4