Amitriptyline and fluoxetine protect PC12 cells from cell death induced by hydrogen peroxide

Amitriptyline and fluoxetine protect PC12 cells from cell death induced by hydrogen peroxide

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J Psychiatry Neurosci 2005;30(3):196-201

Nathan Kolla, MA; Zelan Wei, MD, PhD; J. Steven Richardson, PhD; Xin-Min Li, MD, PhD

Kolla, Wei, Richardson, Li — Neuropsychiatry Research Unit, Department of Psychiatry, University of Saskatchewan; Richardson — Department of Pharmacology, University of Saskatchewan, Saskatoon, Sask.

Abstract


Objective:
To investigate the potential protective effects of amitriptyline and fluoxetine in a catecholamine cell model.

Methods: Cultured rat pheochromocytoma (PC12) cells were pretreated with amitriptyline or fluoxetine for 24 or 48 hours and were then subjected to neurotoxic insult (200 μmol/L hydrogen peroxide). Cell viability was determined by measurement of the reduction product of 3-[4,5-dimethylthiazol-2-yl]-2,5-iphenyltetrazolium bromide (MTT). The enzyme activity of superoxide dismutase (SOD) was determined by a
commercial SOD assay kit.

Results: The decrease in cell viability induced by hydrogen peroxide was attenuated in PC12 cells pretreated with 100 μmol/L amitriptyline for 24 hours or with 50 μmol/L amitriptyline or 50 μmol/L fluoxetine for 48 hours. Pretreatment with either amitriptyline or fluoxetine was associated with increased SOD activity in PC12 cells. Inhibition of SOD activity with diethyldithiocarbamic acid reduced the cytoprotective action of fluoxetine.

Conclusions: These data suggest that the neuroprotective actions of some antidepressants include the upregulation of SOD activity.

Résumé

Objectif : Étudier les effets protecteurs possibles de l’amitriptyline et de la fluoxétine dans un modèle cellulaire de catécholamines.

Méthodes : On a appliqué à des cellules cultivées de phéochromocytome (PC12) de rat un prétraitement de 24 ou de 48 heures à l’amitriptyline ou à la fluoxétine, puis on a induit une atteinte neurotoxique (200 μmol/L de peroxyde d’hydrogène). La viabilité des cellules a été établie par la mesure du produit de réduction du bromure de 3-[4,5-diméthylthiazol-2-yl]-2,5-diphényl-tétrazolium (MTT). L’activité enzymatique de la superoxide dismutase (SOD) a été établie au moyen d’une trousse du commerce pour le dosage de la SOD.

Résultats : Le prétraitement des cellules de PC12 au moyen de 100 μmol/L d’amitriptyline pendant 24 heures ou de 50 μmol/L d’amitriptyline ou de fluoxétine pendant 48 heures a atténué la diminution de la viabilité des cellules causée par le peroxyde d’hydrogène. Le prétraitement aussi bien à l’amitriptyline qu’à la fluoxétine a été associé avec une augmentation de l’activité enzymatique de la SOD dans les cellules de PC12. L’inhibition de l’activité de la SOD au moyen de l’acide diéthyldithiocarbamique a réduit l’action de cytoprotection de la fluoxétine.

Conclusions : Ces données semblent indiquer que l’action de neuroprotection de certains antidépresseurs comprend une régulation à la hausse de l’activité de la SOD.


Medical subject headings: amitriptyline; fluoxetine; PC12 cells; cell viability; superoxide dismutase; oxidative stress.

Submitted Jan. 15, 2004; Revised June 18, 2004; Accepted Aug. 30, 2004

Acknowledgements: We thank Dr. A.V. Juorio for his valuable discussions. This work was supported by the Canadian Psychiatric Research Foundation and the Saskatchewan Health Research Foundation.

Competing interests: None declared.

Contributors: The study was conceived by Drs. Kolla and Wei, and all authors participated in the experimental design. Drs. Kolla and Wei were responsible for data acquisition. All authors participated in data interpretation and drafting the article, and gave final approval for the version to be published.

Correspondence to: Dr. Xin-Min Li, Neuropsychiatry Research Unit, Medical Research Building, University of Saskatchewan, 103 Wiggins Rd., Saskatoon SK S7N 5E4; fax 306 966-8830; xin-min.li@usask.ca