Effect of Glutathione Depletion on Nrf2/ARE Activation by Deltamethrin in PC12 Cells

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Transcription factor NF-E2-related factor 2 (Nrf2) is important for cell protection against chemical-induced oxidative stress. Previously, we have reported that in PC12 cells, Nrf2 can be triggered by deltamethrin (DM), a commonly used pyrethroid insecticide. Molecular mechanisms behind Nrf2 activation by DM are still unclear. Here we studied the effects of cell glutathione (GSH) depletion on Nrf2 activation by DM. We found that DM enhanced Nrf2 expression at the mRNA and protein levels and increased nuclear Nrf2 levels. Activation of Nrf2 was associated with activation of its downstream targets, such as heme oxygenase-1 (HO-1) and glutamate cysteine ligase catalytic subunit (GCLC). In contrast, DL-buthionine-[S,R]- sulfoximine (BSO), a known GSH-depleting agent, did not increase Nrf2 protein expression or cause its nuclear accumulation. However, pre-treatment with BSO triggered mRNA expression of HO-1 and GCLC. Furthermore, BSO pre-treatment suppressed DM-induced Nrf2 upregulation and activation and lowered mRNA expression of HO-1 and GCLC upon DM treatment. These data demonstrate that GSH depletion is not necessary for the activation of Nrf2/ARE by DM in PC12 cells, and that GCLC and HO-1 expression can increase through other signalling pathways.

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  • 1. Casida JE Gammon DW Glickman AH Lawrence LJ. Mechanisms of selective action of pyrethroid insecticides. Annu Rev Pharmacol Toxicol 1983;23:413-38.

  • 2. Soderlund DM Clark JM Sheet LP Mullin LS Piccirillo VJ Sargent D Stevens JT Weiner ML. Mechanisms of pyrethroid neurotoxicity: implications for cumulative risk assessment. Toxicology 2002;171:3-59.

  • 3. Nguyen T Sherratt PJ Pickett CB. Regulatory mechanisms controlling gene expression mediated by the antioxidant response element. Annu Rev Pharmacol Toxicol 2003;43:233-60.

  • 4. Ishii T Itoh K Yamamoto M. Roles of Nrf2 in activation of antioxidant enzyme genes via antioxidant responsive elements. Methods Enzymol 2002;348:182-90.

  • 5. Wild AC Moinova HR Mulcahy RT. Regulation of gammaglutamylcysteine synthetase subunit gene expression by the transcription factor Nrf2. J Biol Chem 1999;274:33627-36.

  • 6. Chan K Kan YW. Nrf2 is essential for protection against acute pulmonary injury in mice. Proc Natl Acad Sci USA 1999;96:12731-6.

  • 7. Chanas SA Jiang Q McMahon M McWalter GK McLellan LI Elcombe CR Henderson CJ Wolf CR Moffat GJ Itoh K Yamamoto M Hayes JD. Loss of the Nrf2 transcription factor causes a marked reduction in constitutive and inducible expression of the glutathione S-transferase Gsta1 Gsta2 Gstm1 Gstm2 Gstm3 and Gstm4 genes in the livers of male and female mice. Biochem J 2002;365:405-16.

  • 8. Alam J Stewart D Touchard C Boinapally S Choi AM Cook JL. Nrf2 a Cap’n’Collar transcription factor regulates induction of the heme oxygenase-1 gene. J Biol Chem 1999;274:26071-8.

  • 9. Wasserman WW Fahl WE. Functional antioxidant responsive elements. Proc Natl Acad Sci USA 1997;94:5361-6.

  • 10. Thimmulappa RK Mai KH Srisuma S Kensler TW Yamamoto M Biswal S. Identification of Nrf2-regulated genes induced by the chemopreventive agent sulforaphane by oligonucleotide microarray. Cancer Res 2002;62:5196-203.

  • 11. Kwak MK Akabayashi N Itoh K Motohash H Yamamoto M Kensler TW. Modulation of gene expression by cancer chemopreventive dithiolethiones through the Keap1-Nrf2 pathway: identification of novel gene clusters for cell survival. J Biol Chem 2003;278:8135-45.

  • 12. Li HY Wu SY Ma Q Shi N. The pesticide deltamethrin increases free radical production and promotes nuclear translocation of the stress response transcription factor Nrf2 in rat brain. Toxicol Ind Health 2011;27:579-90.

  • 13. Li HY Shi N Chen D Dai ZH Lu WH Wang B Li YR. [Oxidative stress of deltamethrin on rats nervous system in Chinese]. Chin J Ind Hyg Occup Dis 2005;23:97-101.

  • 14. Hong F Sekhar KR Freeman ML Liebler DC. Specific patterns of electrophile adduction trigger Keap1 ubiquitination and Nrf2 activation. J Biol Chem 2005;280:31768-75.

  • 15. Kobayashi A Kang M Watai Y Tong K Shibata T Uchida K Yamamoto M. Oxidative and electrophilic stresses activate Nrf2 through inhibition of ubiquitination activity of Keap1. Mol Cell Biol 2006;26:221-9.

  • 16. Zhang DD Lo SC Cross JV Templeton DJ Hannink M. Keap1 is a redox-regulated substrate adaptor protein for a Cul3-dependent ubiquitin ligase complex. Mol Cell Biol 2004;24:10941-53.

  • 17. Huang HC Nguyen T Pickett CB. Phosphorylation of Nrf2 at Ser-40 by protein kinase C regulates antioxidant response element-mediated transcription. J Biol Chem 2002;277:42769-74.

  • 18. Bloom DA Jaiswal AK. Phosphorylation of Nrf2 at Ser40 by protein kinase C in response to antioxidants leads to the release of Nrf2 from INrf2 but is not required for Nrf2 stabilization/accumulation in the nucleus and transcriptional activation of antioxidant response element-mediated NAD(P)H:quinone oxidoreductase-1 gene expression. J Biol Chem 2003;278:44675-82.

  • 19. Jain AK Bloom DA Jaiswal AK. Nuclear import and export signals in control of NRF2. J Biol Chem 2005;280:29158-68.

  • 20. Li W Jain MR Chen C Yue X Hebbar V Zhou R Kong AN. NRF2 possesses a Redox-insensitive nuclear export signal overlapping with the leucine zipper motif. J Biol Chem 2005:280:28430-8.

  • 21. Goldring CE Kitteringham NR Elsby R Randle LE Clement YN Williams DP McMahon M Hayes JD Itoh K Yamamoto M Park BK. Activation of hepatic Nrf2 in vivo by acetaminophen in CD-1 mice. Hepatology 2004;39:1267-76.

  • 22. Timothy JS William DA. Transmitter ion channel and receptor properties of pheochromocytoma (PC12) cells: a model for neurotoxicological studies. NeuroToxicology 1991;12:473-92.

  • 23. Tyrrell RM Pidoux M. Endogenous glutathione protects human skin fibroblasts against the cytotoxic action of UVB UVA and near-visible radiations. Photochem Photobiol 1986;44:561-4.

  • 24. Shen DX Fu JL Shi X Zhou ZC. Toxic effect of inhibition of glutathione synthesis on human embryonic kidney cells. J Health Toxicol 2002;16:150-3.

  • 25. Li HY Wu SY Shi N. Transcription factor Nrf2 activation by deltamethrin in PC12 cells: involvement of ROS. Toxicol Lett 2007;171:87-98.

  • 26. Deptala A Bedner E Gorczyca W Darzynkiewicz Z. Activation of nuclear factor kappa B (NF-κB) assayed by laser scanning cytometry (LSC). Cytometry 1998;33:376-82.

  • 27. Schreiber E Matthias P Mūller MM Schaffner W. Rapid detection of octamer binding proteins with ‘mini-extracts’ prepared from a small number of cells. Nucleic Acids Res 1989;17:6419.

  • 28. Yan CC Huxtable RJ. Fluorimetric determination of monobromobimane and o-phthalaldehyde adducts of gammaglutamylcysteine and glutathione: application to assay of gamma-glutamylcysteinyl synthetase activity and glutathione concentration in liver. J Chromatogr B Biomed Appl 1995;672:217-24.

  • 29. Lee JM Moehlenkamp JD Hanson JM Johnson JA. Nrf2- dependent activation of the antioxidant responsive element by ter-butylhydroquinone is independent of oxidative stress in IMR-32 Human neuroblastoma cells. Biochem Biophysic Res Commun 2001;280:286-92.

  • 30. Shukla GS Chiu J Hart BA. Enhanced expression of pulmonary gamma-glutamylcysteine synthetase heavy subunit in rats exposed to cadmium aerosols. Toxicol Appl Pharmacol 2000;163:249-59.

  • 31. Itoh KWakabayashi N Katoh Y Ishii T Igarashi K Engel JD Yamamoto M. Keap1 represses nuclear activation of antioxidant responsive elements by Nrf2 through binding to the amino-terminal Neh2 domain. Genes Dev 1999;13:76-86.

  • 32. Nguyen T Sherratt PJ Huang H-C Yang CS Pickett CB. Increased protein stability as a mechanism that enhances Nrf2-mediated transcriptional activation of the antioxidant response element: degradation of Nrf2 by the 26 S proteasome. J Biol Chem 2003;278:4536-41.

  • 33. Randle LE Goldring CE Benson CA Metcalfe PN Kitteringham NR Park BKWilliams DP.Investigation of the effect of a panel of model hepatotoxins on the Nrf2-Keap1 defence response pathway in CD-1 mice.Toxicology 2008;20:243:249-60.

  • 34. Zhang H Court N Forman HJ. Submicromolar concentrations of 4-hydroxynonenal induce glutamate cysteine ligase expression in HBE1 cells. Redox Rep 2007;12:101-6.

  • 35. Li HY Shi N Zhong YF. [Effect of deltamethrin on production of reactive oxygen species in PC12 cells in Chinese]. Chin J Ind Hyg Occup Dis 2008;26:654-8.

  • 36. Martin D Rojo AI Salinas M Diaz R Gallardo G Alam J De Galarreta CM Cuadrado A. Regulation of heme oxygenase-1 expression through the phosphatidylinositol 3-kinase/Akt pathway and the Nrf2 transcription factor in response to the antioxidant phytochemical carnosol. J Biol Chem 2004;279:8919-29.

  • 37. Chan K Han XD Kan YW. An important function of Nrf2 in combating oxidative stress: detoxification of acetaminophen. Proc Natl Acad Sci USA 2001;98:4611-6.

  • 38. Hayes JD Chanas SA Henderson CJ McMahon M Sun C Moffat GJ Wolf CR Yamamoto M. The Nrf2 transcription factor contributes both to the basal expression of glutathione S-transferases in mouse liver and to their induction by the chemopreventive synthetic antioxidants butylated hydroxyanisole and ethoxyquin. Biochem Soc Trans 2000;28:33-41.

  • 39. Ishii T Itoh K Takahashi S Sato H Yanagawa T Katoh Y Bannai S Yamamoto M. Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages. J Biol Chem 2000;275;16023-9.

  • 40. Itoh K Chiba T Takahashi S Ishii T Igarashi K Katoh Y Oyake T Hayashi N Satoh K Hatayama I Yamamoto M Nabeshima Y. An Nrf2/small Maf heterodimer mediates the induction of phase II detoxifying enzyme genes through antioxidant response elements. Biochem Biophys Res Commun 1997;236:313-22.

  • 41. McMahon M Itoh K Yamamoto M Chanas SA Henderson CJ McLellan LI Wolf CR Cavin C Hayes JD. The Cap’n’Collar basic leucine zipper transcription factor Nrf2 (NF-E2 p45-related factor 2) controls both constitutive and inducible expression of intestinal detoxification and glutathione biosynthetic enzymes. Cancer Res 2001;61:3299-307.

  • 42. Oguro T Yoshida T Numazawa S Kuroiwa Y. Possible role of glutatione depletion in the induction of rate-limiting enzymes involved in heme degradation and polyamine biosynthesis in the liver of rats. J Pharmacobiodyn 1990;13:628-36.

  • 43. Saunders EL Maines MD Meredith MJ Freeman ML. Enhancement of heme oxygenase 1 synthesis by glutathione depletion in Chinese hamster ovary cells. Arch Biochem Biophys 1991;288:368-73.

  • 44. Lautier D Luscher P Tyrrell RM. Endogenous glutathione level modulate both constitutive and UVA radiation/hydrogen peroxide inducible expression of the human heme oxygenase gene. Carcinogenesis 1992;13:227-32.

  • 45. Ewing JF Maines MD. Glutathione depletion induces heme oxygenase-1 (HSP32) mRNA and protein in rat brain. J Nerochem 1993;60:1512-9.

  • 46. Oguro T Hayashi M Numazawa S Asakawa K Yoshida T. Heme oxygenase-1 gene expression by a glutathione depletor phorone mediated through AP-1 activation in rats. Biochem Biophys Res Commun 1996;221:259-65.

  • 47. Alam J. Heme oxygenase-1: past present and future. Antiox Redox Signal 2002;4:559-62.

  • 48. Alam J Cook JL. Transcriptional regulation of the heme oxygenase-1 gene via the stress response element pathway. Curr Pharm Des 2003;9:2499-511.

  • 49. Balogun E Hoque M Gong P Killeen E Green CJ Foresti R Alam J Motterlini R. Curcumin activates the haem oxygenase-1 gene via regulation of Nrf2 and the antioxidantresponsive element. Biochem J 2003;371:887-95.

  • 50. Naughton P Hoque M Green CJ Foresti R Motterlini R. Interaction of heme with nitroxyl or nitric oxide amplifies heme oxygenase-1 induction: involvement of the transcription factor Nrf2. Cell Mol Biol 2002;48:885-94.

  • 51. Shih AY Johnson DA Wong G Kraft AD Jiang L Erb H Johnson JA Murphy TH. Coordinate regulation of glutathione biosynthesis and release by Nrf2-expressing glia potently protects neurons from oxidative stress. J Neurosci 2003;23:3394-406.

  • 52. Calabrese V Scapagnini G Ravagna A Colombrita C Spadaro F Butterfield DA Giuffrida Stella AM. Increased expression of heat shock proteins in rat brain during aging: relationship with mitochondrial function and glutathione redox state. Mech Ageing Dev 2004;125:325-35.

  • 53. Tian L Shi MM Forman HJ. Increased transcription of the regulatory subunit of γ - glutamylcysteine synthetase in rat lung epithelial L2 cells exposed to oxidative stress or glutathione depletion. Arch Biochem Biophy 1997;342:126-33.

  • 54. Mao LJ Xu HW. Effect of glutathione depletion on glutathione system in myocadium of rat. Chin J Sports Med 2004;23:266-70.

  • 55. Urata Y Yamamoto H Goto S Tsushima H Akazawa S Yamashita S Nagataki S Kondo T. Long exposure to high glucose concentration impairs the responsive expression of gamma-glutamylcysteine synthetase by interleukin-1beta and tumor necrosis factor-alpha in mouse endothelial cells. J Biol Chem 1996;271:15146-52.

  • 56. Gipp JJ Bailey HH Mulcahy RT. Cloning and sequencing of the cDNA for the light subunit of human liver gammaglutamylcysteine synthetase and relative mRNA levels for heavy and light subunits in human normal tissues. Biochem Biophys Res Commun 1995;206:584-9.

  • 57. Wild AC Mulcahy RT. Regulation of gamma-glutamylcysteine synthetase subunit gene expression: insights into transcriptional control of antioxidant defenses. Free Radic Res 2000;32:281-301.

  • 58. Venugopal R Jaiswal AK. Nrf1 and Nrf2 positively and c-Fos and Fra1 negatively regulate the human antioxidant response element-mediated expression of NAD(P)H:quinone oxidoreductase1 gene. Proc Natl Acad Sci USA 1996;93:14960-5.

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