title: Inhibition by R(+) or S(-) pramipexole of caspase activation and cell death induced by methylpyridinium ion or beta amyloid peptide in SH-SY5Y neuroblastoma.
abstract: Cell models of neurodegenerative diseases (NDD) can involve expression of mutant nuclear genes associated with Mendelian forms of the diseases or effects of toxins believed to replicate essential disease features. Death produced by exposing neural cells to methylpyridinium ion (MPP(+)) or neurotoxic beta amyloid (BA) peptides is frequently used to study features of the sporadic, most prevalent forms of Parkinson's disease (PD) and Alzheimer's disease (AD), respectively. We examined in replicating SH-SY5Y human neuroblastoma cells the release of cytochrome C into cytoplasm, activation of caspases 9 and 3, and loss of calcein retention as markers of the "mitochondrial" pathway of cell death. Exposure to 5 mM MPP(+), which induces apoptotic cell death within 18-24 hr, released cytochrome C within 4 hr, activated caspases 9 and 3, and reduced calcein accumulation. BA 25-35 peptide produced more rapid and greater elevations of caspase 3 activity; no effects were observed with the nontoxic BA 35-25 reverse sequence. The dependence on mitochondrial transition pore (MTP) activity of MPP(+)-induced caspase activations was demonstrated by preincubation with bongkreckic acid, which blocked elevations of caspases 9 and 3. Stereoisomers of pramipexole (PPX), a free radical scavenger and inhibitor of MTP opening, inhibited caspase activation (MPP(+) and BA) and restored calcein accumulation (MPP(+)). Our results demonstrate that MPP(+) and BA can induce cell death through MTP-dependent activation of caspase cascades. PPX stereoisomers interfere with activation of these cell death pathways and may be useful clinically as neuroprotectants in PD and AD and related diseases.
authors.affiliation: Center for the Study of Neurodegenerative Diseases, University of Virginia, Charlottesville, VA 22908. bennett@virginia.edu
authors.forename: Nicole A
authors.initials: NA
authors.lastname: Abramova
authors.forename: David S
authors.initials: DS
authors.lastname: Cassarino
authors.forename: Shaharyar M
authors.initials: SM
authors.lastname: Khan
authors.forename: Terry W
authors.initials: TW
authors.lastname: Painter
authors.forename: James P
authors.initials: JP
authors.lastname: Bennett
mesh_terms: D015655:1-Methyl-4-phenylpyridinium / Q000037:antagonists & inhibitors*; D000544:Alzheimer Disease / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D016229:Amyloid beta-Peptides / Q000037:antagonists & inhibitors*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D034341:Aristolochic Acids; D052160:Benzothiazoles; D001865:Bongkrekic Acid / Q000494:pharmacology; D001921:Brain / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects* / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016166:Free Radical Scavengers / Q000494:pharmacology*; D006801:Humans; D009447:Neuroblastoma; D019636:Neurodegenerative Diseases / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D010300:Parkinson Disease / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D010446:Peptide Fragments / Q000037:antagonists & inhibitors; D010616:Phenanthrenes / Q000494:pharmacology; D000077487:Pramipexole; D013844:Thiazoles / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
chemical_list: D016229:Amyloid beta-Peptides; D000900:Anti-Bacterial Agents; D034341:Aristolochic Acids; D052160:Benzothiazoles; D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D010446:Peptide Fragments; D010616:Phenanthrenes; D013844:Thiazoles; C074636:amyloid beta-protein (25-35); D001865:Bongkrekic Acid; D000077487:Pramipexole; C000228:aristolochic acid I; D020169:Caspases; D015655:1-Methyl-4-phenylpyridinium