BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
권호 표지
DOI QR코드

DOI QR Code

The hypertension drug, verapamil, activates Nrf2 by promoting p62-dependent autophagic Keap1 degradation and prevents acetaminophen-induced cytotoxicity

  • Lee, Da Hyun (Severance Biomedical Science Institute, Yonsei Biomedical Research Institute, Yonsei University College of Medicine) ;
  • Park, Jeong Su (Severance Biomedical Science Institute, Yonsei Biomedical Research Institute, Yonsei University College of Medicine) ;
  • Lee, Yu Seol (Severance Biomedical Science Institute, Yonsei Biomedical Research Institute, Yonsei University College of Medicine) ;
  • Sung, Su Haeng (Severance Biomedical Science Institute, Yonsei Biomedical Research Institute, Yonsei University College of Medicine) ;
  • Lee, Yong-ho (Division of Endocrinology and Metabolism, Department of Internal Medicine, Yonsei University College of Medicine) ;
  • Bae, Soo Han (Severance Biomedical Science Institute, Yonsei Biomedical Research Institute, Yonsei University College of Medicine)
  • Received : 2016.11.10
  • Accepted : 2016.12.13
  • Published : 2017.02.28
Download PDF
⟨ Previous Next ⟩

Abstract

Nuclear factor erythroid 2-related factor 2 (Nrf2) provides a cellular defense against oxidative stress by inducing the expression of antioxidant and detoxification enzymes. The calcium antagonist, verapamil, is an FDA-approved drug prescribed for the treatment of hypertension. Here, we show that verapamil acts as a potent Nrf2 activator without causing cytotoxicity, through degradation of Kelch-like ECH-associated protein 1 (Keap1), a Nrf2 repressor. Furthermore, verapamil-induced Keap1 degradation is prominently mediated by a p62-dependent autophagic pathway. Correspondingly, verapamil protects cells from acetaminophen-induced oxidative damage through Nrf2 activation. These results demonstrated the underlying mechanisms for the protective role of verapamil against acetaminophen-induced cytotoxicity.

Keywords

References

  1. Kensler TW, Wakabayashi N and Biswal S (2007) Cell survival responses to environmental stresses via the Keap1-Nrf2-ARE pathway. Annu Rev Pharmacol Toxicol 47, 89-116 https://doi.org/10.1146/annurev.pharmtox.46.120604.141046
  2. Itoh K, Mimura J and Yamamoto M (2010) Discovery of the negative regulator of Nrf2, Keap1: a historical overview. Antioxid Redox Signal 13, 1665-1678 https://doi.org/10.1089/ars.2010.3222
  3. Taguchi K, Motohashi H and Yamamoto M (2011) Molecular mechanisms of the Keap1-Nrf2 pathway in stress response and cancer evolution. Genes Cells 16, 123-140 https://doi.org/10.1111/j.1365-2443.2010.01473.x
  4. Itoh K, Wakabayashi N, Katoh Y et al (1999) Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain. Genes Dev 13, 76-86 https://doi.org/10.1101/gad.13.1.76
  5. Bae SH, Sung SH, Oh SY et al (2013) Sestrins activate Nrf2 by promoting p62-dependent autophagic degradation of Keap1 and prevent oxidative liver damage. Cell Metab 17, 73-84 https://doi.org/10.1016/j.cmet.2012.12.002
  6. Bataille AM and Manautou JE (2012) Nrf2: a potential target for new therapeutics in liver disease. Clin Pharmacol Ther 92, 340-348 https://doi.org/10.1038/clpt.2012.110
  7. Musso G, Cassader M and Gambino R (2016) Nonalcoholic steatohepatitis: emerging molecular targets and therapeutic strategies. Nat Rev Drug Discov 15, 249-274 https://doi.org/10.1038/nrd.2015.3
  8. Johnson JA, Johnson DA, Kraft AD et al (2008) The Nrf2-ARE pathway: an indicator and modulator of oxidative stress in neurodegeneration. Ann N Y Acad Sci 1147, 61-69 https://doi.org/10.1196/annals.1427.036
  9. Zoja C, Benigni A and Remuzzi G (2014) The Nrf2 pathway in the progression of renal disease. Nephrol Dial Transplant 29 Suppl 1, i19-i24 https://doi.org/10.1093/ndt/gft224
  10. Goligorsky MS, Chaimovitz C, Rapoport J, Goldstein J and Kol R (1985) Calcium metabolism in uremic nephrocalcinosis: preventive effect of verapamil. Kidney Int 27, 774-779 https://doi.org/10.1038/ki.1985.79
  11. Rosei EA, Dal Palu C, Leonetti G, Magnani B, Pessina A and Zanchetti A (1997) Clinical results of the Verapamil inHypertension and Atherosclerosis Study. VHAS Investigators. J Hypertens 15, 1337-1344 https://doi.org/10.1097/00004872-199715110-00019
  12. Pajak B, Kania E, Gajkowska B and Orzechowski A (2012) Verapamil-induced autophagy-like process in colon adenocarcinoma COLO 205 cells; the ultrastructural studies. Pharmacol Rep 64, 991-996 https://doi.org/10.1016/S1734-1140(12)70896-4
  13. Park HW, Park H, Semple IA et al (2014) Pharmacological correction of obesity-induced autophagy arrest using calcium channel blockers. Nat Commun 5, 4834 https://doi.org/10.1038/ncomms5834
  14. Park HW and Lee JH (2014) Calcium channel blockers as potential therapeutics for obesity-associated autophagy defects and fatty liver pathologies. Autophagy 10, 2385-2386 https://doi.org/10.4161/15548627.2014.984268
  15. James LP, Mayeux PR and Hinson JA (2003) Acetaminophen-induced hepatotoxicity. Drug Metab Dispos 31, 1499-1506 https://doi.org/10.1124/dmd.31.12.1499
  16. Ni HM, Williams JA, Yang H, Shi YH, Fan J and Ding WX (2012) Targeting autophagy for the treatment of liver diseases. Pharmacol Res 66, 463-474 https://doi.org/10.1016/j.phrs.2012.07.003
  17. Copple IM, Goldring CE, Jenkins RE et al (2008) The hepatotoxic metabolite of acetaminophen directly activates the Keap1-Nrf2 cell defense system. Hepatology 48, 1292-1301 https://doi.org/10.1002/hep.22472
  18. Li N, Alam J, Venkatesan MI et al (2004) Nrf2 is a key transcription factor that regulates antioxidant defense in macrophages and epithelial cells: protecting against the proinflammatory and oxidizing effects of diesel exhaust chemicals. J Immunol 173, 3467-3481 https://doi.org/10.4049/jimmunol.173.5.3467
  19. Taguchi K, Fujikawa N, Komatsu M et al (2012) Keap1 degradation by autophagy for the maintenance of redox homeostasis. Proc Natl Acad Sci U S A 109, 13561-13566 https://doi.org/10.1073/pnas.1121572109
  20. Komatsu M, Kurokawa H, Waguri S et al (2010) The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1. Nat Cell Biol 12, 213-223
  21. Ichimura Y, Waguri S, Sou YS et al (2013) Phosphorylation of p62 activates the Keap1-Nrf2 pathway during selective autophagy. Mol Cell 51, 618-631 https://doi.org/10.1016/j.molcel.2013.08.003
  22. Hinson JA, Roberts DW and James LP (2010) Mechanisms of acetaminophen-induced liver necrosis. Handb Exp Pharmacol 369-405
  23. McGill MR, Sharpe MR, Williams CD, Taha M, Curry SC and Jaeschke H (2012) The mechanism underlying acetaminophen-induced hepatotoxicity in humans and mice involves mitochondrial damage and nuclear DNA fragmentation. J Clin Invest 122, 1574-1583 https://doi.org/10.1172/JCI59755
  24. Goldring CE, Kitteringham NR, Elsby R et al (2004) Activation of hepatic Nrf2 in vivo by acetaminophen in CD-1 mice. Hepatology 39, 1267-1276 https://doi.org/10.1002/hep.20183
  25. Chan K, Han XD and Kan YW (2001) An important function of Nrf2 in combating oxidative stress: detoxification of acetaminophen. Proc Natl Acad Sci U S A 98, 4611-4616 https://doi.org/10.1073/pnas.081082098
  26. McMahon M, Thomas N, Itoh K, Yamamoto M and Hayes JD (2006) Dimerization of substrate adaptors can facilitate cullin-mediated ubiquitylation of proteins by a "tethering" mechanism: a two-site interaction model for the Nrf2-Keap1 complex. J Biol Chem 281, 24756-24768 https://doi.org/10.1074/jbc.M601119200
  27. Yu Z, Shao W, Chiang Y et al (2011) Oltipraz upregulates the nuclear factor (erythroid-derived 2)-like 2 [corrected] (NRF2) antioxidant system and prevents insulin resistance and obesity induced by a high-fat diet in C57BL/6J mice. Diabetologia 54, 922-934 https://doi.org/10.1007/s00125-010-2001-8
  28. Katsuragi Y, Ichimura Y and Komatsu M (2015) p62/SQSTM1 functions as a signaling hub and an autophagy adaptor. FEBS J 282, 4672-4678 https://doi.org/10.1111/febs.13540
  29. Lau A, Wang XJ, Zhao F et al (2010) A noncanonical mechanism of Nrf2 activation by autophagy deficiency: direct interaction between Keap1 and p62. Mol Cell Biol 30, 3275-3285 https://doi.org/10.1128/MCB.00248-10
  30. Lee DH, Han DH, Nam KT et al (2016) Ezetimibe, an NPC1L1 inhibitor, is a potent Nrf2 activator that protects mice from diet-induced nonalcoholic steatohepatitis. Free Radic Biol Med 99, 520-532 https://doi.org/10.1016/j.freeradbiomed.2016.09.009
  31. Sarkar S (2013) Regulation of autophagy by mTORdependent and mTOR-independent pathways: autophagy dysfunction in neurodegenerative diseases and therapeutic application of autophagy enhancers. Biochem Soc Trans 41, 1103-1130 https://doi.org/10.1042/BST20130134
  32. Schmieder RE, Messerli FH, Garavaglia GE and Nunez BD (1987) Cardiovascular effects of verapamil in patients with essential hypertension. Circulation 75, 1030-1036 https://doi.org/10.1161/01.CIR.75.5.1030
  33. Prescott LF and Critchley JA (1983) The treatment of acetaminophen poisoning. Annu Rev Pharmacol Toxicol 23, 87-101 https://doi.org/10.1146/annurev.pa.23.040183.000511
  34. Jiang J, Briede JJ, Jennen DG et al (2015) Increased mitochondrial ROS formation by acetaminophen in human hepatic cells is associated with gene expression changes suggesting disruption of the mitochondrial electron transport chain. Toxicol Lett 234, 139-150 https://doi.org/10.1016/j.toxlet.2015.02.012
  35. Liu J, Wu KC, Lu YF, Ekuase E and Klaassen CD (2013) Nrf2 protection against liver injury produced by various hepatotoxicants. Oxid Med Cell Longev 2013, 305861

Cited by

  1. Strange Bedfellows: Nuclear Factor, Erythroid 2-Like 2 (Nrf2) and Hypoxia-Inducible Factor 1 (HIF-1) in Tumor Hypoxia vol.6, pp.2, 2017, https://doi.org/10.3390/antiox6020027
  2. vol.175, pp.3, 2018, https://doi.org/10.1111/bph.14093
  3. Activation of the Nrf2/HO-1 Signaling Pathway Contributes to the Protective Effects of Sargassum serratifolium Extract against Oxidative Stress-Induced DNA Damage and Apoptosis in SW1353 Human Chondrocytes vol.15, pp.6, 2018, https://doi.org/10.3390/ijerph15061173
  4. Comparative effects of calcium and potassium channel modulators on ischemia/reperfusion injury in the isolated rat heart pp.1573-4919, 2018, https://doi.org/10.1007/s11010-018-3384-y
  5. Role of p62 in the regulation of cell death induction vol.23, pp.3-4, 2018, https://doi.org/10.1007/s10495-018-1445-z