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Protective effects of noradrenaline on benzo[a]pyrene-induced oxidative stress responses in brain tumor cell lines

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Abstract

Benzo[a]pyrene (B[a]P) is an ubiquitous environmental pollutant that is generated during combustion of fossil fuels. We examine the effect of noradrenaline (NA) on B[a]P-induced neurotoxicity in brain tumor cell lines like neuroblastoma (Neuro2a) and glioma (C6). We pre-treated tumor cells with NA for 6 h, followed by addition of B[a]P for additional 24 h. Cell viability was measured using trypan blue dye-exclusion assay and comet assay was performed to measure DNA damage. Cell cycle status was analyzed using flow cytometry and oxidative DNA damage (8-oxodG) production was examined by immunostaining. The intracellular Ca2+ concentration was analyzed using Fura-2AM. Our results showed viability of Neuro2a and C6 cells declined (24% and 20%) in B[a]P-treated groups. However, pre-treating with NA increased viability of cells by reducing percentage of cell death in both. Furthermore, B[a]P-induced deregulation of cell cycle (G2/M and S phase cell arrest) was significantly restored by pre-treatment with NA in Neuro2a cells as compared to C6 cells. We further observed increased 8-oxodG production in B[a]P-treated cells; however, NA pre-treatment significantly (p < 0.05) reduced the 8-oxodG production in Neuro2a, while C6 cells were less affected possibly due to better protective machinery. B[a]P-induced intracellular Ca2+ influx was significantly reduced in both the cell lines due to co-treatment of NA possibly by reducing Ca2+ influx. NA protects brain tumor cells against B[a]P-induced neurotoxicity may be by decreasing percentage of G2 cell arrest, oxidative DNA damage, and reducing intracellular Ca2+ influx. These findings suggested that NA may be considered as a natural potential protective agent against B[a]P-induced neurotoxicity.

Graphical abstract showing differential protective mechanism of NA against B[a]P-induced toxicity through antioxidant mechanism maintaining homeostasis for oxidative stress in Neuro2a and C6 cell lines. The schematic graph showed the biological significance of the NA that regulates the induction of metabolic processes of cell cycle after exposure to the environmental pollutants. B[a]P increases the intracellular levels of Ca2+, but also induces damage to cellular molecules including DNA causing cell cycle arrest. The B[a]P-induced DNA damage due to base lesions generated in the genome, 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) is one of the most abundant because of guanine’s lowest redox potential among DNA bases through intracellular calcium homoeostasis

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References

  • ATSDR (1995) Toxicological profile for polycyclic aromatic hydrocarbons (PAHs). U.S. Department of Health and Human Services, Atlanta

    Google Scholar 

  • Briede JJ, Godschalk RW, Emans MT, De Kok TM, Van Agen E, Van Maanen J et al (2004) In vitro and in vivo studies on oxygen free radical and DNA adduct formation in rat lung and liver during benzo[a]pyrene metabolism. Free Radic Res 38(9):995–1002

    Article  CAS  PubMed  Google Scholar 

  • Das G, Gopalakrishnan A, Faisal M, Mallick BN (2008) Stimulatory role of calcium in rapid eye movement sleep deprivation-induced noradrenaline-mediated increase in Na-K-ATPase activity in rat brain. Neuroscience 155(1):76–89

    Article  CAS  PubMed  Google Scholar 

  • Dutta K, Ghosh D, Nazmi A, Kumawat KL, Basu A (2010) A common carcinogen benzo[a]pyrene causes neuronal death in mouse via microglial activation. PLoS One 5(4):e9984

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Garg R, Gupta S, Maru GB (2008) Dietary curcumin modulates transcriptional regulators of phase I and phase II enzymes in benzo[a]pyrene-treated mice: mechanism of its anti-initiating action. Carcinogenesis 29(5):1022–1032

    Article  CAS  PubMed  Google Scholar 

  • Iimori K, Tanaka M, Kohno Y, Ida Y, Nakagawa R, Hoaki Y, Tsuda A, Nagasaki N (1982) Psychological stress enhances noradrenaline turnover in specific brain regions in rats. Pharmacol Biochem Behav 16(4):637–640

    Article  CAS  PubMed  Google Scholar 

  • Jayasekara S, Sharma RP, Drown DB (1992) Effects of benzo[a]pyrene on steady-state levels of biogenic amines and metabolizing enzymes in mouse brain regions. Ecotoxicol Environ Saf 24(1):1–12

    Article  CAS  PubMed  Google Scholar 

  • Kawashima T, Ohkubo K, Fukuzumi S (2010) Radical scavenging reactivity of catecholamine neurotransmitters and the inhibition effect for DNA cleavage. J Phys Chem B 114(1):675–680

    Article  CAS  PubMed  Google Scholar 

  • LeBel CP, Bondy SC (1991) Oxygen radicals: common mediators of neurotoxicity. Neurotoxicol Teratol 13:341–346

    Article  CAS  PubMed  Google Scholar 

  • Mc Kinnon PJ (2009) DNA repair deficiency and neurological disease. Nat Rev Neurosci 10:100–112

    Article  CAS  Google Scholar 

  • Michel PP, Vyas S, Anglade P, Ruberg M, Agid Y (1994) Morphological and molecular characterization of the response of differentiated PC12 cells to calcium stress. Eur J Neurosci 6(4):577–586

    Article  CAS  PubMed  Google Scholar 

  • Nebert DW, Roe Al, Dieter MZ, Solis WA, Wang Y, Dalto TP (2002) Role of the aromatic hydrocarbon receptor (Ah) gene battery in the oxidative stress response cell cycle control and apoptosis. Biochem Pharmacol 5965–5985

  • Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi CA (1991) Rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139(2):271–279

    Article  CAS  PubMed  Google Scholar 

  • Nwagbara O, Darling-Reed SF, Tucker A, Harris C, Abazinge M, Thomas RD et al (2007) Induction of cell death, DNA strand breaks, and cell cycle arrest in DU145 human prostate carcinoma cell line by benzo[a]pyrene and benzo[a]pyrene-7,8-diol-9,10- epoxide. Int J Environ Res Public Health 4(1):10–14

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ordway GA, Schwartz MA, Frazer A (eds) (2007) Brain noradrenaline neurobiology and therapeutics. 1 ed. Cambridge University Press, New York

    Google Scholar 

  • Patel B, Das SK, Patri M (2016) Neonatal benzo[a]pyrene exposure induces oxidative stress and DNA damage causing neurobehavioural changes during the early adolescence period in rats. Dev Neurosci 38(2):150–162

  • Patri M, Singh A, Mallick BN (2013) Protective role of noradrenaline in Benzo[a]pyrene-induced learning impairment in developing rat. J Neurosci Res 91:1450–1462

    Article  CAS  PubMed  Google Scholar 

  • Pifl C, Zezula J, Spittler A, Kattinger A, Reither H, Caron MG, Hornykiewicz O (2001) Antiproliferative action of dopamine and norepinephrine in neuroblastoma cells expressing the human dopamine transporter. FASEB J 15(9):1607–1609

    Article  CAS  PubMed  Google Scholar 

  • Saunders CR, Das SK, Ramesh A, Shockley DC, Mukherjee S (2006) Benzo[a]pyrene-induced acute neurotoxicity in the F-344 rat: role of oxidative stress. J Appl Toxicol 26:427–438

    Article  CAS  PubMed  Google Scholar 

  • Saunders CR, Shockley DC, Knuckles ME (2001) Behavioral effects induced by acute exposure to benzo(a)pyrene in F-344 rats. Neurotox Res 3(6):557–579

    Article  CAS  PubMed  Google Scholar 

  • Stipursky J, Romao L, Tortelli V, Neto VM, Gomes FC (2011) Neuron-glia signaling: implications for astrocyte differentiation and synapse formation. Life Sci 89(15–16):524–531

    Article  CAS  PubMed  Google Scholar 

  • Sullivan PD (1985) Free radicals of benzo(a)pyrene and derivatives. Environ Health Perspect 64:283–295

    Article  CAS  PubMed  Google Scholar 

  • Takamasa I, Yumi T, Koichi S, Masaki M, Rintaro Y, Kayo Y, Hiromi O, Noboru K, Munehiro N, Yorihiro Y, Phil SH, Naoaki I (2017) Endogenous reactive oxygen species cause astrocyte defects and neuronal dysfunctions in the hippocampus: a new model for aging brain. Aging Cell 16:39–51

    Article  CAS  Google Scholar 

  • Troadec JD, Marien M, Darios F, Hartmann A, Ruberg M, Colpaert F et al (2001) Noradrenaline provides long-term protection to dopaminergic neurons by reducing oxidative stress. J Neurochem 79(1):200–210

    Article  CAS  PubMed  Google Scholar 

  • Troadec JD, Marien M, Mourlevat S, Debeir T, Ruberg M, Colpaert F, Michel PP (2002) Activation of the mitogen-activated protein kinase (ERK(1/2)) signaling pathway by cyclic AMP potentiates the neuroprotective effect of the neurotransmitter noradrenaline on dopaminergic neurons. Mol Pharmacol 62(5):1043–1052

    Article  CAS  PubMed  Google Scholar 

  • Uberti D, Ferrari, Toninelli G, Memo M (2003) Involvement of DNA damage and repair systems in neurodegenerative process. Toxicol Lett 139(2–3):99–105

    Article  CAS  PubMed  Google Scholar 

  • Voorhess ML, Gardner LI (1961) Urinary excretion of norepinephrine, epinephrine and 3-methoxy-4-hydroxymandelic acid by children with neuroblastoma. J Clin Endocrinol Metab 21:321–335

    Article  CAS  PubMed  Google Scholar 

  • Xia Y, Cheng S, He J, Liu X, Tang Y, Yuan H, He L, Lu T, Tu B, Wang Y (2011) Effects of subchronic exposure to benzo[a]pyrene (B[a]P) on learning and memory, and neurotransmitters in male Sprague-Dawley rat. Neurotoxicology. 32(2):188–198

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors also thank all members of Neurobiology Laboratory, School of Life Sciences, JNU, New Delhi, for their kind help, technical support, and instrumentation facility.

Funding

This work was supported by funding from the Project DAE-BRNS, Mumbai, No. 37(1)14/27/2015/ BRNS and DRDO, New Delhi, No. O/o DG (TM)/81/48222/LSRB-294/PEE&BS/2017 to Manorama Patri.

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  1. Neurobiology Laboratory, Department of Zoology, School of Life Sciences, Ravenshaw University, Cuttack, 753003, India

    Manorama Patri & Abhisek Singh

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  1. Manorama Patri
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  2. Abhisek Singh
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Correspondence to Manorama Patri.

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The authors declare that they have no competing interests.

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Editor: Tetsuji Okamoto

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Patri, M., Singh, A. Protective effects of noradrenaline on benzo[a]pyrene-induced oxidative stress responses in brain tumor cell lines. In Vitro Cell.Dev.Biol.-Animal 55, 665–675 (2019). https://doi.org/10.1007/s11626-019-00378-9

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  • DOI: https://doi.org/10.1007/s11626-019-00378-9

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