Neurotoxicity of amphetamine derivatives is mediated by caspase pathway activation in rat cerebellar granule cells.
The neurotoxic action of the abuse drugs methamphetamine (METH) and 3,4-methylenedioxymethamphetamine (MDMA) on cerebellar granule neurones (CGNs) culture was examined. Treatment for 48 h with METH or MDMA (1-5 mM) induced a higher decrease in viability than 24 h treatment. z.VAD.fmk (100 microM) but not MK-801 nor NBQX recovered control viability values. In both cases, cell death was characterised as apoptotic rather than necrotic by morphology cell observation. Apoptosis measured by flow cytometry indicated an increase in the hypodiploid population after 48 h treatment with METH and MDMA. Apoptosis was reverted by the presence of z.VAD.fmk (100 microM) but not by 10 microM MK-801 or NBQX. Similar results were obtained by analysing nuclear chromatine condensation. These results ruled out excitotoxic participation in amphetamine derivative-induced neurotoxicity in CGNs. Participation of radical oxygen species (ROS) was evaluated using alpha-tocopherol (1-15 microM) and cytometric studies. The co-treatment with 4 mM METH or MDMA for 48 h partially reverted neurotoxic action and apoptotic features, indicating ROS implication in CGNs death by amphetamine derivatives. Alteration of mitochondrial function induced cytochrome C (Cyt C) release after 48-h treatment with METH and MDMA (4 mM). There was also indication of caspase-3-like activation, measured by immunoanalysis and biochemically. Finally, neurodegenerative action caused by amphetamine derivatives may be prevented by using caspase inhibitors.
Jiménez
Andrés
A
Unitat de Farmacologia i Farmacognòsia, Facultat de Farmàcia, Universitat de Barcelona, Barcelona 08028, Spain.
Jordà
Elvira G
EG
Verdaguer
Ester
E
Pubill
David
D
Sureda
Francesc X
FX
Canudas
Anna M
AM
Escubedo
Elena
E
Camarasa
Jordi
J
Camins
Antoni
A
Pallàs
Mercè
M
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology* / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004347:Drug Interactions; D004789:Enzyme Activation; D005434:Flow Cytometry; D008694:Methamphetamine / Q000037:antagonists & inhibitors* / Q000633:toxicity*; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D008928:Mitochondria / Q000201:enzymology / Q000378:metabolism; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000037:antagonists & inhibitors* / Q000633:toxicity*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology*; D011810:Quinoxalines / Q000494:pharmacology; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D024502:alpha-Tocopherol / Q000494:pharmacology
D000590:Amino Acid Chloromethyl Ketones; D018696:Neuroprotective Agents; D011810:Quinoxalines; D017382:Reactive Oxygen Species; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C062865:2,3-dioxo-6-nitro-7-sulfamoylbenzo(f)quinoxaline; D008694:Methamphetamine; D016291:Dizocilpine Maleate; D045304:Cytochromes c; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D024502:alpha-Tocopherol; D018817:N-Methyl-3,4-methylenedioxyamphetamine