Monoamine neurotoxins-induced apoptosis in lymphocytes by a common oxidative stress mechanism: involvement of hydrogen peroxide (H(2)O(2)), caspase-3, and nuclear factor kappa-B (NF-kappaB), p53, c-Jun transcription factors.
The destruction of dopaminergic and serotonergic nerve cells by selective 6-hydroxydopamine (6-OHDA), 5,6-dihydroxytryptamine (5,6-DHT) and 5,7-dihydroxytryptamine (5,7-DHT), respectively, is a commonly used tool to investigate the mapping of neuronal pathways, elucidation of function and to mimic human neurodegenerative disease such as Parkinson's and Alzheimer's diseases. Despite intense investigations, a complete picture of the precise molecular cascade leading to cell death in a single cellular model is still lacking. In this study, we provide evidence that 6-OHDA, 5,6- and 5,7-DHT toxins-induced apoptosis in peripheral blood lymphocytes cells in a concentration-dependent fashion by a common oxidative mechanism involving: (1) the oxidation of toxins into quinones and production of the by-product hydrogen peroxide, reflected by desipramine-a monoamine uptake blocker-and antioxidants inhibition, (2) activation and/or translocation of nuclear factor-kappaB, p53 and c-Jun transcription factors, showed by immunocytochemical diaminobenzidine-positive stained nuclei, (3) caspase-3 activation, reflected by caspase Ac-DEVD-CHO inhibition, (4) mRNA and protein synthesis de novo according to cycloheximide and actinomycin D cell death inhibition. These results are consistent with the notion that uptake and intracellular autoxidation of those toxins precede the apoptotic process and that once H(2)O(2) is generated, it is able to trigger a specific cell death signalisation. Thus, taken together these results, we present an ordered cascade of the major molecular events leading peripheral blood lymphocytes to apoptosis. These results may contribute to explain the importance of H(2)O(2) as a second messenger of death signal in some degenerative diseases linked to oxidative stress stimuli.
Del Rio
Marlene Jimenez
MJ
Department of Internal Medicine, School of Medicine, University of Antioquia, Calle 62 no. 52-72, P.O. Box 1226, Medellin, Colombia. mdelrio@quimbaya.udea.edu.co
Velez-Pardo
Carlos
C
D015115:5,6-Dihydroxytryptamine / Q000494:pharmacology; D015116:5,7-Dihydroxytryptamine / Q000494:pharmacology; D018663:Adrenergic Agents / Q000494:pharmacology; D000328:Adult; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D003891:Desipramine / Q000494:pharmacology; D015259:Dopamine Agents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004587:Electrophoresis, Agar Gel; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D066298:In Vitro Techniques; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects*; D008297:Male; D016328:NF-kappa B / Q000378:metabolism; D009498:Neurotoxins / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology; D016627:Oxidopamine / Q000494:pharmacology; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D018490:Serotonin Agents / Q000494:pharmacology; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D018663:Adrenergic Agents; D000975:Antioxidants; D015259:Dopamine Agents; D016328:NF-kappa B; D009498:Neurotoxins; D016755:Proto-Oncogene Proteins c-jun; D018490:Serotonin Agents; D016159:Tumor Suppressor Protein p53; D015116:5,7-Dihydroxytryptamine; D016627:Oxidopamine; D006861:Hydrogen Peroxide; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D003891:Desipramine; D015115:5,6-Dihydroxytryptamine