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Stabilization of Nrf2 by tBHQ prevents LPS-induced apoptosis in differentiated PC12 cells

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Abstract

The inflammatory reaction plays an important role in the pathogenesis of the neurodegenerative disorders. tert-butylhydroquinone (tBHQ) exhibits a wide range of pharmacological activities including anti-oxidative and anti-inflammatory action. In this study, we tried to elucidate possible effects of tBHQ on lipopolysaccharide (LPS)-induced inflammatory reaction and its underlying mechanism in neuron-like PC12 cells. tBHQ inhibited LPS-induced generation of reactive oxygen species (ROS) and elevation of intracellular calcium level. It also inhibited LPS-induced cyclooxygenase 2 (COX-2), TNF-α, nuclear factor KappaB (NF-kB), and caspase-3 expression in a dose-dependent manner while stabilizing nuclear factor-erythroid 2 p45-related factor 2. Moreover, the phosphorylations of p38, ERK1/2, and JNK were suppressed by tBHQ. These results suggest that the anti-inflammatory properties of tBHQ might result from inhibition of COX-2 and TNF-α expression, inhibition of NF-kB nuclear translocation along with suppression of MAP kinases (p38, ERK1/2, and JNK) phosphorylation in PC12 cells, so may be a useful agent for prevention of inflammatory diseases.

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Abbreviations

Aβ:

Amyloid β

AREs:

Antioxidant responsive elements

CAT:

Catalase

CNS:

Central nervous system

ECL:

Electrochemiluminescence

GSH:

Glutathione

H2O2 :

Hydrogen peroxide

Keap1:

Kelch-like ECH-associated protein 1

MDA:

Malondialdehyde

MTT:

3-[4,5-Dimethylthiazol-2-yl]-2, 5-dephenyl tetrazolium bromide

NF-κB:

Nuclear factor-κB

NGF:

Nerve growth factor

Nrf2:

Nuclear factor-erythroid 2 p45-related factor 2

ROS:

Reactive oxygen species

SOD:

Superoxide dismutase

tBHQ:

tert-Butylhydroquinone

References

  1. Shimohama S (2000) Apoptosis in Alzheimer’s disease—an update. Apoptosis 5(1):9–16

    Article  CAS  PubMed  Google Scholar 

  2. Ashe KH (2001) Learning and memory in transgenic mice modeling Alzheimer’s disease. Learn Mem 8(6):301–308

    Article  CAS  PubMed  Google Scholar 

  3. Yang F, Sun X, Beech W, Teter B, Wu S, Sigel J, Vinters HV, Frautschy SA, Cole GM (1998) Antibody to caspase-cleaved actin detects apoptosis in differentiated neuroblastoma and plaque-associated neurons and microglia in Alzheimer’s disease. Am J Pathol 152(2):379–389

    CAS  PubMed  Google Scholar 

  4. Irizarry MC, Hyman BT (2001) Alzheimer disease therapeutics. J Neuropathol Exp Neurol 60(10):923–928

    CAS  PubMed  Google Scholar 

  5. Guha M, Mackman N (2001) LPS induction of gene expression in human monocytes. Cell Signal 13(2):85–94

    Article  CAS  PubMed  Google Scholar 

  6. Allan SM, Rothwell NJ (2003) Inflammation in central nervous system injury. Philos Trans R Soc Lond B Biol Sci 358(1438):1669–1677

    Article  CAS  PubMed  Google Scholar 

  7. Li W, Khor TO, Xu C, Shen G, Jeong WS, Yu S, Kong AN (2008) Activation of Nrf2-antioxidant signaling attenuates NFkappaB-inflammatory response and elicits apoptosis. Biochem Pharmacol 76(11):1485–1489

    Article  CAS  PubMed  Google Scholar 

  8. Nair S, Xu C, Shen G, Hebbar V, Gopalakrishnan A, Hu R, Jain MR, Liew C, Chan JY, Kong AN (2007) Toxicogenomics of endoplasmic reticulum stress inducer tunicamycin in the small intestine and liver of Nrf2 knockout and C57BL/6 J mice. Toxicol Lett 168(1):21–39

    Article  CAS  PubMed  Google Scholar 

  9. Khodagholi F, Eftekharzadeh B, Maghsoudi N, Rezaei PF (2010) Chitosan prevents oxidative stress-induced amyloid beta formation and cytotoxicity in NT2 neurons: involvement of transcription factors Nrf2 and NF-kappaB. Mol Cell Biochem 337(1–2):39–51

    Article  CAS  PubMed  Google Scholar 

  10. Stewart D, Killeen E, Naquin R, Alam S, Alam J (2003) Degradation of transcription factor Nrf2 via the ubiquitin-proteasome pathway and stabilization by cadmium. J Biol Chem 278(4):2396–2402

    Article  CAS  PubMed  Google Scholar 

  11. Tusi SK, Ansari N, Amini M, Amirabad AD, Shafiee A, Khodagholi F (2010) Attenuation of NF-κB and activation of Nrf2 signaling by 1,2,4-triazine derivatives, protects neuron-like PC12 cells against apoptosis. Apoptosis 15:738–751

    Article  CAS  PubMed  Google Scholar 

  12. Koh K, Cha Y, Kim S, Kim J (2009) tBHQ inhibits LPS-induced microglial activation via Nrf2-mediated suppression of p38 phosphorylation. Biochem Biophys Res Commun 380(3):449–453

    Article  CAS  PubMed  Google Scholar 

  13. Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  14. Kutuk O, Basaga H (2003) Aspirin prevents apoptosis and NF-kappaB activation induced by H2O2 in HeLa cells. Free Radic Res 37:1267–1276

    Article  CAS  PubMed  Google Scholar 

  15. Ellman GL (1959) Tissue sulfhydryl groups. Arch Biochem Biophys 82:70–77

    Article  CAS  PubMed  Google Scholar 

  16. Draper HH, Hadley M (1990) Malondialdehyde determination as index of lipid peroxidation. Methods Enzymol 186:421–431

    Article  CAS  PubMed  Google Scholar 

  17. Kakkar P, Das B, Viswanathan PN (1984) A modified spectrophotometric assay of superoxide dismutase. Ind J Biochem Biophys 21:130–132

    CAS  Google Scholar 

  18. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126

    Article  CAS  PubMed  Google Scholar 

  19. Guroff G (1985) PC12 cells as a model of neuronal differentiation. In: Bottenstein JE (ed) Cell culture in neurosciences. Plenum Press, New York, pp 245–272

    Google Scholar 

  20. Martin TF, Grishanin RN (2003) PC12 cells as a model for studies of regulated secretion in neuronal and endocrine cells. Methods Cell Biol 71:267–286

    Article  CAS  PubMed  Google Scholar 

  21. Omata Y, Saito Y, Fujita K, Ogawa Y, Nishio K, Yoshida Y, Niki E (2008) Induction of adaptive response and enhancement of PC12 cell tolerance by lipopolysaccharide primarily through the upregulation of glutathione S-transferase A3 via Nrf2 activation. Free Radic Biol Med 45:1437–1445

    Article  CAS  PubMed  Google Scholar 

  22. Miller DK (1997) The role of caspase family of cysteine proteases in apoptosis. Semin Immunol 9:35–49

    Article  CAS  PubMed  Google Scholar 

  23. Yu LC, Turner JR, Buret AG (2006) LPS/CD14 activation triggers SGLT-1-mediated glucose uptake and cell rescue in intestinal epithelial cells via early apoptotic signals upstream of caspase-3, Exp. Cell Res 312:3276–3286

    Article  CAS  Google Scholar 

  24. Eftekharzadeh B, Maghsoudi N, Khodagholi F (2010) Stabilization of transcription factor Nrf2 by tBHQ prevents oxidative stress-induced amyloid beta formation in NT2 N neurons. Biochimie 92(3):245–253

    Article  CAS  PubMed  Google Scholar 

  25. Li J, Johnson D, Calkins M, Wright L, Svendsen C, Johnson J (2005) Stabilization of Nrf2 by tBHQ confers protection against oxidative stress-induced cell death in human neural stem cells. Toxicol Sci 83(2):313–328

    Article  CAS  PubMed  Google Scholar 

  26. Liu CS, Chen NH, Zhang JT (2007) Protection of PC12 cells from hydrogen peroxide-induced cytotoxicity by salvianolic acid B, a new compound isolated from Radix Salviae miltiorrhizae. Phytomedicine 14(7–8):492–497

    Article  CAS  PubMed  Google Scholar 

  27. Blum D, Torch S, Nissou MF, Verna JM (2001) 6-Hydroxydopamine-induced nuclear factor-kappa B activation in PC12 cells. Biochem Pharmacol 62(4):473–481

    Google Scholar 

  28. Cárcamo JM, Pedraza A, Bórquez-Ojeda O, Zhang B, Sanchez R, Golde DW (2004) Vitamin C is a kinase inhibitor: dehydroascorbic acid inhibits IkappaBalpha kinase beta. Mol Cell Biol 24(15):6645–6652

    Article  PubMed  Google Scholar 

  29. Shishodia S, Potdar P, Gairola CG, Aggarwal BB (2003) Curcumin (diferuloylmethane) down-regulates cigarette smoke-induced NF-kappaB activation through inhibition of IkappaBalpha kinase in human lung epithelial cells: correlation with suppression of COX-2, MMP-9 and cyclin D1. Carcinogenesis 24(7):1269–1279

    Article  CAS  PubMed  Google Scholar 

  30. Li Y, Liu L, Barger SW, Mrak RE, Griffin WS (2001) Vitamin E suppression of microglial activation is neuroprotective. J Neurosci Res 66(2):163–170

    Article  CAS  PubMed  Google Scholar 

  31. Scapagnini G, Colombrita C, Amadio M, D’Agata V, Arcelli E, Sapienza M, Quattrone A, Calabrese V (2006) Curcumin activates defensive genes and protects neurons against oxidative stress. Antioxid Redox Signal 8:395–403

    Article  CAS  PubMed  Google Scholar 

  32. Greene LA, Tischler AS (1976) Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci USA 73(7):2424–2428

    Article  CAS  PubMed  Google Scholar 

  33. Cowley S, Paterson H, Kemp P, Marshall CJ (1994) Activation of MAP kinase kinase is necessary and sufficient for PC12 differentiation and for transformation of NIH 3T3 cells. Cell 77(6):841–852

    Article  CAS  PubMed  Google Scholar 

  34. Heneka MT, Löschmann PA, Gleichmann M, Weller M, Schulz JB, Wüllner U, Klockgether T (1998) Induction of nitric oxide synthase and nitric oxide-mediated apoptosis in neuronal PC12 cells after stimulation with tumor necrosis factor-alpha/lipopolysaccharide. J Neurochem 71(1):88–94

    Article  CAS  PubMed  Google Scholar 

  35. Männel DN, Rosenstreich DL, Mergenhagen SE (1979) Mechanism of lipopolysaccharide-induced tumor necrosis: requirement for lipopolysaccharide-sensitive lymphoreticular cells. Infect Immun 24(2):573–576

    PubMed  Google Scholar 

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

    Article  CAS  Google Scholar 

  37. Nguyen T, Nioi P, Pickett CB (2009) The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem 284(20):13291–13295

    Article  CAS  PubMed  Google Scholar 

  38. He X, Lin GX, Chen MG, Zhang JX, Ma Q (2007) Protection against chromium (VI)-induced oxidative stress and apoptosis by Nrf2. Recruiting Nrf2 into the nucleus and disrupting the nuclear Nrf2/Keap1 association. Toxicol Sci 98(1):298–309

    Article  CAS  PubMed  Google Scholar 

  39. Ho HK, White CC, Fernandez C, Fausto N, Kavanagh TJ, Nelson SD, Bruschi SA (2005) Nrf2 activation involves an oxidative-stress independent pathway in tetrafluoroethylcysteine-induced cytotoxicity. Toxicol Sci 86(2):354–364

    Article  CAS  PubMed  Google Scholar 

  40. Jiang T, Huang Z, Chan JY, Zhang DD (2009) Nrf2 protects against As(III)-induced damage in mouse liver and bladder. Toxicol Appl Pharmacol 240(1):8–14

    Article  CAS  PubMed  Google Scholar 

  41. Kamata H, Honda S, Maeda S, Chang L, Hirata H, Karin M (2005) Reactive oxygen species promote TNFalpha-induced death and sustained JNK activation by inhibiting MAP kinase phosphatases. Cell 120(5):649–661

    Article  CAS  PubMed  Google Scholar 

  42. Karin M (2006) Nuclear factor-kappaB in cancer development and progression. Nature 441(7092):431–436

    Article  CAS  PubMed  Google Scholar 

  43. Papa S, Bubici C, Zazzeroni F, Pham CG, Kuntzen C, Knabb JR, Dean K, Franzoso G (2006) The NF-kappaB-mediated control of the JNK cascade in the antagonism of programmed cell death in health and disease. Cell Death Differ 13(5):712–729

    Article  CAS  PubMed  Google Scholar 

  44. Pham CG, Bubici C, Zazzeroni F, Papa S, Jones J, Alvarez K, Jayawardena S, De Smaele E, Cong R, Beaumont C, Torti FM, Torti SV, Franzoso G (2004) Ferritin heavy chain upregulation by NF-kappaB inhibits TNFalpha-induced apoptosis by suppressing reactive oxygen species. Cell 119(4):529–542

    Article  CAS  PubMed  Google Scholar 

  45. Sakon S, Xue X, Takekawa M, Sasazuki T, Okazaki T, Kojima Y, Piao JH, Yagita H, Okumura K, Doi T, Nakano H (2003) NF-kappaB inhibits TNF-induced accumulation of ROS that mediate prolonged MAPK activation and necrotic cell death. EMBO J 22(15):3898–3909

    Article  CAS  PubMed  Google Scholar 

  46. Claudio E, Brown K, Siebenlist U (2006) NF-kappaB guides the survival and differentiation of developing lymphocytes. Cell Death Differ 13(5):697–701

    Article  CAS  PubMed  Google Scholar 

  47. Hayden MS, Ghosh S (2004) Signaling to NF-kappaB. Genes Dev 18(18):2195–2224

    Article  CAS  PubMed  Google Scholar 

  48. Kucharczak J, Simmons MJ, Fan Y, Gélinas C (2003) To be, or not to be: NF-kappaB is the answer–role of Rel/NF-kappaB in the regulation of apoptosis. Oncogene 22(56):8961–8982

    Article  CAS  PubMed  Google Scholar 

  49. Gilmore TD (2006) Introduction to NF-kappaB: players, pathways, perspectives. Oncogene 25(51):6680–6684

    Article  CAS  PubMed  Google Scholar 

  50. Pearson G, Robinson F, Beers Gibson T, Xu BE, Karandikar M, Berman K, Cobb MH (2001) Mitogen-activated protein (MAP) kinase pathways: regulation and physiological functions. Endocr Rev 22(2):153–183

    Article  CAS  PubMed  Google Scholar 

  51. Davis RJ (2000) Signal transduction by the JNK group of MAP kinases. Cell 103(2):239–252

    Article  CAS  PubMed  Google Scholar 

  52. Kyriakis JM, Avruch J (2001) Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev 81(2):807–869

    CAS  PubMed  Google Scholar 

  53. Ramachandiran S, Huang Q, Dong J, Lau SS, Monks TJ (2002) Mitogen-activated protein kinases contribute to reactive oxygen species-induced cell death in renal proximal tubule epithelial cells. Chem Res Toxicol 15(12):1635–1642

    Article  CAS  PubMed  Google Scholar 

  54. Zanotto-Filho A, Delgado-Cañedo A, Schröder R, Becker M, Klamt F, Moreira JC (2010) The pharmacological NFkappaB inhibitors BAY117082 and MG132 induce cell arrest and apoptosis in leukemia cells through ROS-mitochondria pathway activation. Cancer Lett 288(2):192–203

    Article  CAS  PubMed  Google Scholar 

  55. Aggarwal BB, Natarajan K (1996) Tumor necrosis factors: developments during the last decade. Eur Cytokine Netw 7(2):93–124

    CAS  PubMed  Google Scholar 

  56. Locksley RM, Killeen N, Lenardo MJ (2001) The TNF and TNF receptor superfamilies: integrating mammalian biology. Cell 104(4):487–501

    Article  CAS  PubMed  Google Scholar 

  57. Wang H, Joseph JA (2000) Mechanisms of hydrogen peroxide-induced calcium dysregulation in PC12 cells. Free Radic Biol Med 28(8):1222–1231

    Article  CAS  PubMed  Google Scholar 

  58. Tan S, Sagara Y, Liu Y, Maher P, Schubert D (1998) The regulation of reactive oxygen species production during programmed cell death. J Cell Biol 141(6):1423–1432

    Article  CAS  PubMed  Google Scholar 

  59. Castillo MR, Babson JR (1998) Ca(2+)-dependent mechanisms of cell injury in cultured cortical neurons. Neuroscience 86(4):1133–1144

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by Shahid Beheshti University of Medical Sciences Research Funds. The authors thank MS. Joodi for her help in preparing the manuscript.

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Authors and Affiliations

  1. Neuroscience Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Fariba Khodagholi & Solaleh Khoramian Tusi

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  1. Fariba Khodagholi
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  2. Solaleh Khoramian Tusi
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Correspondence to Fariba Khodagholi.

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Khodagholi, F., Tusi, S.K. Stabilization of Nrf2 by tBHQ prevents LPS-induced apoptosis in differentiated PC12 cells. Mol Cell Biochem 354, 97–112 (2011). https://doi.org/10.1007/s11010-011-0809-2

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