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Neuroprotective Effect of Quercetin Against Hydrogen Peroxide-induced Oxidative Injury in P19 Neurons

Journal of Molecular Neuroscience volume 47, pages 286–299 (2012)Cite this article

Abstract

Oxidative stress is implicated in neuronal death in a variety of neurodegenerative diseases. In the present study, P19 neurons obtained by the differentiation procedure from mouse teratocarcinoma P19 cells were used to investigate the ability of quercetin, a plant-derived flavonoid, to prevent neuronal death induced by exposure to 150 μM or 1.5 mM hydrogen peroxide (H2O2) for 24 h. Quercetin treatment improved viability of P19 neurons exposed to both types of oxidative injury. During the modest oxidative stress, quercetin diminished generation of reactive oxygen species (ROS) and prevented H2O2-induced nuclear condensation, increase in caspase 3/7 activity and rise in poly(APD-ribose) polymerase expression. Expression of Bcl-2 family members Bax and Bcl-2 was not affected by quercetin treatment at both the transcriptional and translational levels. During the severe oxidative injury, quercetin prevented H2O2-induced rise in ROS accumulation and changes in plasma membrane integrity and nuclear morphology. The obtained results suggest that neuroprotective effects of quercetin are related to its antioxidative action and prevention of events associated with programmed cell death cascade. In the light of these findings, one might assume beneficial effects of quercetin for the prevention of oxidative stress-driven neuronal loss in human aging and age-related neurodegenerative diseases.

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Abbreviations

AIF:

Apoptosis-inducing factor

ATRA:

all-trans retinoic acid

BBB:

Blood–brain barrier

DCF-DA:

Dichlorofluorescin diacetate

DIV:

days in vitro

MTT:

3(4,5-Dimethylthiazol-2yl)2,5-dyphenyl-2H-tetrazolium bromide

PARP:

poly (ADP-ribose) polymerase

ROS:

Reactive oxygen species

NAD+ :

Nicotinamide adenine dinucleotide

RFU:

Relative fluorescence unit

References

  • Ahmad A, Khan MM, Hoda MN et al (2011) Quercetin protects against oxidative stress associated damages in a rat model of transient focal cerebral ischemia and reperfusion. Neurochem Res 36:1360–1371

    PubMed  Article  CAS  Google Scholar 

  • Andrabi SA, Dawson TM, Dawson VL (2008) Mitochondrial and nuclear cross talk in cell death. Ann NY Acad Sci 1147:233–241

    Google Scholar 

  • Akhtar RS, Ness JM, Roth KA (2004) Bcl-2 family regulation of neuronal development and neurodegeneration. Biochim Biophys Acta 1644:189–203

    PubMed  Article  CAS  Google Scholar 

  • Arredondo F, Echeverry C, Abin-Carriquiry JA et al (2010) After cellular internalization, quercetin causes Nrf2 nuclear translocation, increases glutathione levels, and prevents neuronal death against an oxidative insult. Free Radic Biol Med 49:738–747

    PubMed  Article  CAS  Google Scholar 

  • Boots AW, Haenen GRMM, Bast A (2008) Health effects of quercetin: from antioxidant to nutraceutical. Eur J Pharmacol 585:325–337

    PubMed  Article  CAS  Google Scholar 

  • Chandra J, Samali A, Orrenius S (2000) Triggering and modulation of apoptosis by oxidative stress. Free Radic Biol Med 29:323–333

    PubMed  Article  CAS  Google Scholar 

  • Chen T-J, Jeng J-Y, Lin C-W, Wu C-Y, Chen Y-C (2006) Quercetin inhibition of ROS-dependent and -independent apoptosis in rat glioma C6 cells. Toxicology 223:113–126

    PubMed  Article  CAS  Google Scholar 

  • Choi EJ, Kim G-H (2010) Quercetin accumulation by chronic administration causes the caspase-3 activation in liver and brain of mice. Biofactors 36:216–221

    PubMed  Article  Google Scholar 

  • Cole KK, Perez-Polo JR (2002) Poly(ADP-ribose) polymerase inhibition prevents both apoptotic-like delayed neuronal death and necrosis after H2O2 injury. J Neurochem 82:19–29

    PubMed  Article  CAS  Google Scholar 

  • Cookson MR, Ince PG, Shaw PJ (1998) Peroxynitrite and hydrogen peroxide induced cell death in the NSC34 neuroblastoma × spinal cord cell line: role of poly(ADP-ribose) polymerase. J Neurochem 70:501–508

    PubMed  Article  CAS  Google Scholar 

  • Crossthwaite AJ, Hasan S, Williams RJ (2002) Hydrogen peroxide-mediated phosphorylation of ERK1/2, Akt/PKB and JNK in cortical neurons: dependence on Ca2+ and PI3-kinase. J Neurochem 80:24–35

    PubMed  Article  CAS  Google Scholar 

  • Dajas F, Rivera-Megret F, Blasina F et al (2003) Neuroprotection by flavonoids. Braz J Med Biol Res 36:1613–1620

    PubMed  Article  CAS  Google Scholar 

  • Das S, Mandal AK, Ghosh A, Panda S, Das N, Sarkar S (2008) Nanoparticulated quercetin in combating age related cerebral oxidative injury. Curr Aging Sci 1:169–174

    PubMed  Article  CAS  Google Scholar 

  • Desagher S, Glowinski J, Premont J (1996) Astrocytes protect neurons from hydrogen peroxide toxicity. J Neurosci 16:2553–2562

    PubMed  CAS  Google Scholar 

  • Dok-Go H, Lee KH, Kim HJ et al (2003) Neuroprotective effects of antioxidative flavonoids, quercetin, (+)-dihydroquercetin and quercetin 3-methyl ether, isolated from Opuntia ficus-indica var. saboten. Brain Res 965:130–136

    PubMed  Article  CAS  Google Scholar 

  • Echeverry C, Arredondo F, Abin-Carriquiry JA (2010) Pretreatment with natural flavones and neuronal cell survival after oxidative stress: a structure–activity relationship study. J Agric Food Chem 58:2111–2115

    PubMed  Article  CAS  Google Scholar 

  • Emerit J, Edeas M, Bricaire F (2004) Neurodegenerative diseases and oxidative stress. Biomed Pharmacother 58:39–46

    PubMed  Article  CAS  Google Scholar 

  • Fatokun AA, Stone TW, Smith RA (2007) Cell death in rat cerebellar granule neurons induced by hydrogen peroxide in vitro: mechanisms and protection by adenosine receptor ligands. Brain Res 1132:193–202

    PubMed  Article  CAS  Google Scholar 

  • Geraets L, Moonen HJJ, Brauers K, Wouters EFM, Bast A, Hageman GJ (2007) Dietary flavones and flavonoles are inhibitors of poly(ADP-ribose)polymerase-1 in pulmonary epithelial cells. J Nutr 137:2190–2195

    PubMed  CAS  Google Scholar 

  • Hampton MB, Orrenius S (1997) Dual regulation of caspase activity by hydrogen peroxide: implications for apoptosis. FEBS Lett 414:552–556

    PubMed  Article  CAS  Google Scholar 

  • Heo HJ, Lee CY (2004) Protective effects of quercetin and vitamin C against oxidative stress-induced neurodegeneration. J Agric Food Chem 52:7514–7517

    PubMed  Article  CAS  Google Scholar 

  • Ishige K, Schubert D, Sagara Y (2001) Flavonoids protect neuronal cells from oxidative stress by three distinct mechanisms. Free Radic Biol Med 30:433–446

    PubMed  Article  CAS  Google Scholar 

  • Ishihara I, Minami Y, Nishizaki T, Matsuoka T, Yamamura H (2000) Activation of calpain precedes morphological alterations during hydrogen peroxide-induced apoptosis in neuronally differentiated mouse embryonal carcinoma P19 cell line. Neurosci Lett 279:97–100

    PubMed  Article  CAS  Google Scholar 

  • Jakubowicz-Gil J, Rzeski W, Zdzisinska B, Dobrowolski P, Gawron A (2008) Cell death and neuronal arborization upon quercetin treatment in rat neurons. Acta Neurobiol Exp 68:139–146

    Google Scholar 

  • Jones-Villeneuve EMV, McBurney MW, Rogers KA, Kalnins VI (1982) Retinoic acid induces embryonal carcinoma cells to differentiate into neurons and glial cells. J Cell Biol 94:253–262

    PubMed  Article  CAS  Google Scholar 

  • Kauppinen TM (2007) Multiple roles for poly(ADP-ribose)polymerase-1 in neurological disease. Neurochem Int 50:954–958

    PubMed  Article  CAS  Google Scholar 

  • Kelsey NA, Wilkins HM, Linseman DA (2010) Nutraceutical antioxidants as novel neuroprotective agents. Molecules 15:7779–7814

    Article  Google Scholar 

  • Krantic S, Mechawar N, Reix S, Quirion R (2005) Molecular basis of programmed cell death involved in neurodegeneration. Trends Neurosci 28:670–676

    PubMed  Article  CAS  Google Scholar 

  • Li G-Y, Osborne NN (2008) Oxidative-induced apoptosis to an immortalized ganglion cell line is caspase independent but involves the activation of poly(ADP-ribose)polymerase and apoptosis inducing factor. Brain Res 1188:35–43

    PubMed  Article  CAS  Google Scholar 

  • Mercer LD, Kelly BL, Horne MK, Beart PM (2005) Dietary polyphenols protect dopamine neurons from oxidative insults and apoptosis: investigations in primary rat mesencephalic cultures. Biochem Pharmacol 69:339–345

    PubMed  Article  CAS  Google Scholar 

  • Moroni F (2008) Poly(ADP-ribose)polymerase 1 (PARP-1) and postischemic brain damage. Curr Opin Pharmacol 8:96–103

    Google Scholar 

  • Oršolić N, Benković V, Lisičić D, Đikić D, Erhardt J, Horvat-Knežević A (2010) Protective effects of propolis and related polyphenolic/flavonoid compounds against toxicity induced by irinotecan. Med Oncol 27:1346–1358

    PubMed  Article  Google Scholar 

  • Oršolić N, Gajski G, Garaj-Vrhovac V, Đikić D, Prskalo ZŠ, Sirovina D (2011) DNA-protective effects of quercetin or naringenin in alloxan-induced diabetic mice. Eur J Pharmacol 656:110–8

    PubMed  Article  Google Scholar 

  • Ossola B, Kääriäinen TM, Männistö PT (2009) The multiple faces of quercetin in neuroprotection. Expert Opin Drug Saf 8:397–409

    PubMed  Article  CAS  Google Scholar 

  • Park C, So H-S, Shin C-H et al (2003) Quercetin protects the hydrogen peroxide-induced apoptosis via inhibition of mitochondrial dysfunction in H9c2 cardiomyoblast cells. Biochem Pharmacol 66:1287–1295

    PubMed  Article  CAS  Google Scholar 

  • Pavlica S, Gebhardt R (2010) Protective effects of flavonoids and two metabolites against oxidative stress in neuronal PC12 cells. Life Sci 86:79–86

    PubMed  Article  CAS  Google Scholar 

  • Ruffels J, Griffin M, Dickenson JM (2004) Activation of ERK1/2, JNK and PKB by hydrogen peroxide in human SH-SY5Y neuroblastoma cells: role of ERK1/2 in H2O2-induced cell death. Eur J Pharmacol 483:163–173

    PubMed  Article  CAS  Google Scholar 

  • Samali A, Nordgren H, Zhivotovsky B, Peterson E, Orrenius S (1999) A comparative study of apoptosis and necrosis in HepG2 cells: oxidant-induced caspase inactivation leads to necrosis. Biochem Biophys Res Commun 255:6–11

    PubMed  Article  CAS  Google Scholar 

  • Slemmer JE, Shacka JJ, Sweeney MI, Weber JT (2008) Antioxidants and free radical scavengers for the treatment of stroke, traumatic brain injury and aging. Curr Med Chem 15:404–414

    PubMed  Article  CAS  Google Scholar 

  • Spencer JPE, Rice-Evans C, Williams RJ (2003) Modulation of pro-survival Akt/protein kinase B and ERK1/2 signaling cascades by quercetin and its in vivo metabolites underlie their action on neuronal viability. J Biol Chem 278:34783–34793

    PubMed  Article  CAS  Google Scholar 

  • Steckley D, Karajgikar M, Dale LB et al (2007) Puma is a dominant regulator of oxidative stress induced Bax activation and neuronal apoptosis. J Neurosci 27:12989–12999

    PubMed  Article  CAS  Google Scholar 

  • Suematsu N, Hosoda M, Fujimori K (2011) Protective effects of quercetin against hydrogen peroxide-induced apoptosis in human neuronal SH-SY5Y cells. Neurosci Lett 504:223–227

    PubMed  Article  CAS  Google Scholar 

  • Sugawara T, Fujimura M, Noshita N et al (2004) Neuronal death/survival signaling pathways in cerebral ischemia. NeuroRx 1:17–25

    PubMed  Article  Google Scholar 

  • Turetsky DM, Huettner JE, Gottlieb DI, Goldberg MP, Choi DW (1993) Glutamate receptor-mediated currents and toxicity in embryonal carcinoma cells. J Neurobiol 24:1157–1169

    PubMed  Article  CAS  Google Scholar 

  • Vafeiadou K, Vauzour D, Rodriguez-Mateos A, Whiteman M, Williams RJ, Spencer JPE (2008) Glial metabolism of quercetin reduces its neurotoxic potential. Arch Biochem Biophys 478:195–200

    PubMed  Article  CAS  Google Scholar 

  • Valencia A, Moran J (2004) Reactive oxygen species induce different cell death mechanisms in cultured neurons. Free Radic Biol Med 36:1112–1125

    PubMed  Article  CAS  Google Scholar 

  • Vargas AJ, Burd R (2010) Hormesis and synergy: pathways and mechanisms of quercetin in cancer prevention and management. Nutr Rev 68:418–428

    PubMed  Article  Google Scholar 

  • Wang H, Joseph JA (1999) Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Rad Biol Med 27:612–616

    PubMed  Article  CAS  Google Scholar 

  • Wang Y, Dawson VL, Dawson TM (2009) Poly(ADP-ribose) signals to mitochondrial AIF: a key event in parthanatos. Exp Neurol 218:193–202

    PubMed  Article  CAS  Google Scholar 

  • Williams RJ, Spencer JPE, Rice-Evans C (2004) Flavonoids: antioxidants or signalling molecules? Free Radic. Biol Med 36:838–849

    CAS  Google Scholar 

  • Yu S-W, Wang H, Poitras MF et al (2002) Mediation of poly(ADP-ribose) polymerase-1-dependent cell death by apoptosis inducing factor. Science 297:259–263

    PubMed  Article  CAS  Google Scholar 

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Acknowledgments

Undifferentiated P19 cells were kindly provided by Dr. J. Pachernik (Prague, Czech Republic). The skillful technical assistance to Sanjica Rak and Zlatica Tonšetić is gratefully acknowledged. The study was supported by the Croatian Ministry of Science, Education and Sport.

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Authors and Affiliations

  1. Laboratory for Molecular Neuropharmacology, Division of Molecular Medicine, Rudjer Boskovic Institute, Bijenicka 54, Zagreb, Croatia

    Maja Jazvinšćak Jembrek

  2. Laboratory for Genotoxic Agents, Division of Molecular Biology, Rudjer Boskovic Institute, Bijenicka 54, Zagreb, Croatia

    Lidija Vuković

  3. Department of Animal Physiology, Faculty of Science, University of Zagreb, Rooseveltov trg 6, Zagreb, Croatia

    Jasmina Puhović, Julija Erhardt & Nada Oršolić

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  1. Maja Jazvinšćak Jembrek
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  2. Lidija Vuković
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  5. Nada Oršolić
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Correspondence to Maja Jazvinšćak Jembrek.

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Jazvinšćak Jembrek, M., Vuković, L., Puhović, J. et al. Neuroprotective Effect of Quercetin Against Hydrogen Peroxide-induced Oxidative Injury in P19 Neurons. J Mol Neurosci 47, 286–299 (2012). https://doi.org/10.1007/s12031-012-9737-1

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  • DOI: https://doi.org/10.1007/s12031-012-9737-1

Keywords

  • Quercetin
  • Oxidative stress
  • H2O2-induced injury
  • P19 neurons
  • Neuroprotection
  • PARP expression