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Neurotoxicity Research

, Volume 19, Issue 4, pp 544–555 | Cite as

Adverse Effects of 2,4-Dichlorophenoxyacetic Acid on Rat Cerebellar Granule Cell Cultures Were Attenuated by Amphetamine

  • B. Bongiovanni
  • A. Ferri
  • A. Brusco
  • M. Rassetto
  • L. M. Lopez
  • A. M. Evangelista de Duffard
  • R. Duffard
Article

Abstract

2,4-Dichlorophenoxyacetic acid (2,4-D), a worldwide-used herbicide, has been shown to produce a wide range of adverse effects in the health—from embryotoxicity and teratogenicity to neurotoxicity—of animals and humans. In this study, neuronal morphology and biochemical events in rat cerebellar granule cell (CGC) cultures have been analyzed to define some of the possible mechanisms involved in 2,4-D-induced cell death. For that purpose, amphetamine (AMPH) that has been shown to accelerate the recovery of several functions in animals with brain injury has been used as a pharmacologycal tool and was also investigated as a possible protecting agent. Addition of 2,4-D to CGC cultures produced a drastic decrease in cell viability, in association with an increased incidence of necrosis and apoptosis, and an increased level of reactive oxygen species, a decrease in glutathione content, and an abnormal activity of some enzymes with respect to the control group. The adverse effects of 2,4-D were partly attenuated in presence of AMPH. Some deleterious effects on several ultrastructural features of the cells, as well as the enhanced incidence of apoptosis, were partially preserved in AMPH-protected cultures as compared with those which were exposed to 2,4-D alone. The collected evidences (1) confirms the previously observed, deleterious effects of 2.4D on the same or a similar model; (2) suggests that the 2,4-D-induced apoptosis could have been mediated by or associated to an oxidative imbalance in the affected cells, and (3) shows some evidence of a protective effect of AMPH on 2,4-D-induced cell death, which could have been exerted through a reduction in the oxidative stress.

Keywords

2,4-D Amphetamine Oxidative stress Apoptosis Protective effect Nerve cell culture 

Abbreviations

2,4-D

2,4-Dichlorophenoxyacetic acid

AMPH

Amphetamine

CAT

Catalase

CGC

Cerebellar granule cells

CNS

Central nervous system

Cu/Zn-SOD

Cu/Zn superoxide dismutase

DCF

2′,7′-Dichlorofluorescein

DCFH

2′,7′-Dichlorfluorescein-diacetate

DMEM

Dulbecco’s modified Eagle’s medium

FBS

Heat-inactivated fetal bovine serum

GSH

Reduced glutathione

HBSS

Hanks balanced salt solution

Mn-SOD

Mn-superoxide dismutase

MTT

3-(4,5 dimethylthiazol-2-eyl)-2,5-diphenyltetrazolium bromide

NO

Nitric oxide

RNS

Reactive nitrogen species

ROS

Reactive oxygen species

Se-GPx

Selenium-glutathione peroxidase

Notes

Acknowledgments

This study was supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) and Agencia de Promoción Científica y Tecnológica, Argentina, to Dr. Ricardo Duffard, and from the University of Buenos Aires (M-072) to Dr. Alicia Brusco. We thank Prof. Mercedes Leiva and Hebe Bottai for assistance in the statistical analysis.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • B. Bongiovanni
    • 1
  • A. Ferri
    • 2
  • A. Brusco
    • 3
  • M. Rassetto
    • 1
  • L. M. Lopez
    • 3
  • A. M. Evangelista de Duffard
    • 1
  • R. Duffard
    • 1
  1. 1.Laboratorio de Toxicología Experimental (LATOEX), Facultad de Ciencias Bioquímicas y FarmacéuticasUniversidad Nacional de RosarioRosarioArgentina
  2. 2.Departamento de Química Analítica, Facultad de Ciencias Bioquímicas y FarmacéuticasUniversidad Nacional de RosarioRosarioArgentina
  3. 3.Instituto Biología Celular y Neurociencias, LANAIS-MIE, Facultad de MedicinaUniversidad de Buenos AiresBuenos AiresArgentina

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