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Melatonin decreases the oxidative stress produced by 2,4-dichlorophenoxyacetic acid in rat cerebellar granule cells

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Abstract

2,4-Dichlorophenoxyacetic acid (2,4-D) is one of the most widely used herbicides due to its relatively moderate toxicity and to its biodegrad-ability in the soil. In toxic concentrations, 2,4- D displays strong neurotoxicity, partly due to generation of free radicals. Since melatonin has remarkable antioxidant properties, the objective of this study was to assess to what extent it was effective in preventing the 2,4-D effect on redox balance of rat cerebellar granule cells (CGC) in vitro. Cellular viability, generation of reactive oxygen species (ROS) and reactive nitrogen species (RNS), reduced glutathione (GSH) levels, and the activities of the antioxidant enzymes Cu/ Zn-superoxide dismutase (Cu/Zn-SOD), Mn-SOD, selenium-glutathione peroxidase (Se-GPx) and catalase (CAT) were measured in CGC exposed to 2,4-D and/or melatonin for 48 h. In CGC cultures exposed to 2,4-D, cell viability, GSH levels and CAT activity decreased significantly whereas ROS generation and Se-GPx activities were augmented. Except for Se-GPx activity, all these changes were counteracted by the concomitant addition of 0.1 or 0.5 mM melatonin. In addition, incubation of CGC with melatonin alone resulted in augmentation of cell viability, GSH levels and Se-GPx activity. RNS generation and SOD activity remained unaffected by either treatment. Since melatonin was able to counteract most of redox changes produced by 2,4-D in CGC in culture, the experimental evidence reported further support the efficacy of melatonin to act as a neuroprotector.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic acid

CAT:

catalase

CGC:

Cerebellar granule cells

CNS:

Central nervous system

Cu/Zn-SOD:

Cu/Zn-superoxide dismutase

DCF:

2′,7′-dichlorofluorescein

DCFH:

2′7′-dichlorofluorescin

GSH:

reduced glutathione

Mn-SOD:

Mn-superoxide dismutase

MTT:

3-(4,5-dimethylthiazol-2-eyl)-2,5-diphenyl tetrazolium bromide

NMDA:

N-methyl-D-aspartate

NO:

nitric oxide

RNS:

reactive nitrogen species

ROS:

reactive oxygen species

Se-GePx:

selenium-glutathione peroxidase

References

  • Beers RF and IW Sizer (1951) A spectrophotometric method for measuring the breakdown of hydrogen peroxide by cata-lase.J. Biol. Chem. 195, 133–140.

    Google Scholar 

  • Bortolozzi A, AM Evangelista de Duffard, F Dajas, R Duffard and R Silveira (2001) Intracerebral administration of 2,4-diclorophenoxyacetic acid induces behavioral and neuro-chemical alterations in the rat brain.Neurotoxicology 22, 221–232.

    Article  PubMed  CAS  Google Scholar 

  • Bradberry SM, BE Watt, AT Proudfoot and JA Vale (2000) Mechanisms of toxicity, clinical features, and management of acute chlorophenoxy herbicide poisoning: a review.J. Toxicol. Clin. Toxicol. 38, 111–122.

    Article  PubMed  CAS  Google Scholar 

  • Brusco A, JP Saavedra, G Garcia, P Tagliaferro, AM Evangelista de Duffard and R Duffard (1997) 2,4-dichlorophenoxyacetic acid through lactation induces astrogliosis in rat brain.Mol. Chem. Neuropathol. 30, 175–185.

    Article  PubMed  CAS  Google Scholar 

  • Cardinali DP, RE Rosenstein, DA Golombek, HE Chuluyan, B Kanterewicz, MM Del Zar and MI Vacas (1991) Melatonin binding sites in brain: single or multiple?Adv. Pineal Res. 5, 159–165.

    CAS  Google Scholar 

  • De Moliner KL, AM Evangelista de Duffard, E Soto, R Duffard and AM Adamo (2002) Induction of apoptosis in cerebellar granule cells by 2,4-dichlorophenoxyacetic acid.Neurochem. Res. 27, 1439–1446.

    Article  PubMed  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Gallo V, R Suergiu and G Levi (1987) Functional evaluation of glutamate receptor subtypes in cultured cerebellar neurones and astrocytes.Eur. J. Pharmacol. 138, 293–297.

    Article  PubMed  CAS  Google Scholar 

  • Giusti P, M Gusella, M Lipartiti, D Milani, W Zhu, S Vicini and H Manev (1995) Melatonin protects primary cultures of cerebellar granule neurons from kainate but not fromN- methyl-D-aspartate excitotoxicity.Exp. Neurol. 131, 39–46.

    Article  PubMed  CAS  Google Scholar 

  • Kaioumova D, C Susal and G Opelz (2001) Induction of apoptosis in human lymphocytes by the herbicide 2,4-dichlorophenoxyacetic acid.Hum. Immunol. 62, 64–74.

    Article  PubMed  CAS  Google Scholar 

  • McCord JM and I Fridovich (1969) The utility of superoxide dismutase in studying free radical reactions. I. Radicals generated by the interaction of sulfite, dimethyl sulfoxide, and oxygen.J. Biol. Chem. 244, 6056–6063.

    PubMed  CAS  Google Scholar 

  • Miranda KM, MG Espey and DA Wink (2001) A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite.Nitric Oxide 5, 62–71.

    Article  PubMed  CAS  Google Scholar 

  • Myhre O, TA Vestad, E Sagstuen, H Aarnes and F Fonnum (2000) The effects of aliphatic (n-nonane), naphtenic (1,2, 4-trimethylcyclohexane), and aromatic (1,2,4-trimethylben-zene) hydrocarbons on respiratory burst in human neutrophil granulocytes.Toxicol. Appl. Pharmacol. 167, 222–230.

    Article  PubMed  CAS  Google Scholar 

  • Paglia DE and WN Valentine (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathi-one peroxidase.J. Lab. Clin. Med. 70, 158–169.

    PubMed  CAS  Google Scholar 

  • Pandi-Perumal SR, V Srinivasan, GJM Maestroni DP Cardinali, B Poeggeler and R Hardeland (2006) Melatonin: nature’s most versatile biological signal?FEBS J. 273, 2813–2838.

    Article  PubMed  CAS  Google Scholar 

  • Park JS, M Wang, SJ Park and SH Lee (1999) Zinc finger of replication protein A, a non-DNA binding element, regulates its DNA binding activity through redox.J. Biol. Chem. 274, 29075–29080.

    Article  PubMed  CAS  Google Scholar 

  • Savill RM, PJ Scotting and B Coyle (2005) Strategies to investigate gene expression and function in granule cells.Cerebellum 4, 271–278.

    Article  PubMed  CAS  Google Scholar 

  • Srinivasan V, SR Pandi-Perumal, GJ Maestroni, AI Esquifino, R Hardeland and DP Cardinali (2005) Role of melatonin in neurodegenerative diseases.Neurotox. Res. 7, 293–318.

    Article  PubMed  CAS  Google Scholar 

  • Teixeira MC, JP Telo, NF Duarte and I Sa-Correia (2004) The herbicide 2,4-dichlorophenoxyacetic acid induces the generation of free-radicals and associated oxidative stress responses in yeast.Biochem. Biophys. Res. Commun. 324, 1101–1107.

    Article  PubMed  CAS  Google Scholar 

  • Thomas T, F Rauscher, R Sanders, J Veltman and JB WatkinsIII (2000) Effects of aldose reductase inhibitors on anti-oxidant defense in rat and rabbit liver.Toxicol. Sci. 53, 145–149.

    Article  PubMed  CAS  Google Scholar 

  • Tricoire H, M Moller, P Chemineau and B Malpaux (2003) Origin of cerebrospinal fluid melatonin and possible function in the integration of photoperiod.Reprod. Suppl. 61, 311–321.

    PubMed  CAS  Google Scholar 

  • Tuschl H and C Schwab (2003) Cytotoxic effects of the herbicide 2,4-dichlorophenoxyacetic acid in HepG2 cells.Food Chem. Toxicol. 41, 385–393.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Laboratorio de Toxicología Experimental (LATOEX) Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina

    B. Bongiovanni, P. De Lorenzi, A. Ferri, C. Konjuh, M. Rassetto, A. M. Evangelista De Duffard & R. Duffard

  2. Departamento de Fisiología, Facultad de Medicina, Universidad de Buenos Aires, Buenos Aires, Argentina

    D. P. Cardinali

Authors
  1. B. Bongiovanni
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  2. P. De Lorenzi
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  3. A. Ferri
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  4. C. Konjuh
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  5. M. Rassetto
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  6. A. M. Evangelista De Duffard
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  7. D. P. Cardinali
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  8. R. Duffard
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Correspondence to D. P. Cardinali.

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Bongiovanni, B., De Lorenzi, P., Ferri, A. et al. Melatonin decreases the oxidative stress produced by 2,4-dichlorophenoxyacetic acid in rat cerebellar granule cells. neurotox res 11, 93–99 (2007). https://doi.org/10.1007/BF03033388

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  • DOI: https://doi.org/10.1007/BF03033388

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