An investigation into the role of reactive oxygen species in the mechanism of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity using neuronal cell lines.
The study of oxygen radical generation and effects during 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) metabolism was undertaken in an in vitro test system. Three neurochemically discrete neuronal cell lines, B50 (cholinergic) and B65 rat cell lines and SKNSH human neuroblastoma (both catecholaminergic), were exposed to MPTP (0-200 microM). Parallel experiments were performed using reagent H2O2, an intermediate which may be generated during MPTP metabolism, to determine whether MPTP and H2O2 had any selectivity of toxicity and whether the mechanisms of cell death were similar. MPTP toxicity was shown to be reduced by monoamine oxidase B inhibitors, pargyline (P < 0.01) and selegiline (P < 0.05), indicating that toxicity was due to metabolism of MPTP rather than the parent compound. Cytotoxicity was also decreased in the presence of antioxidants, most notably in the presence of superoxide dismutase and catalase together (P < 0.01), suggesting that reactive oxygen species (ROS) play a role in MPTP-induced cell death. Attempts to determine the intracellular target for oxidative attack did not identify significant levels of lipid peroxidation products, but did demonstrate nucleoid expansion, possibly the result of double stranded DNA breaks induced by ROS.
Lai
M
M
Wolfson Research Laboratories, Queen Elizabeth Medical Centre, Edgbaston, Birmingham, U.K.
Griffiths
H
H
Pall
H
H
Williams
A
A
Lunec
J
J
D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine / Q000031:analogs & derivatives*; D015655:1-Methyl-4-phenylpyridinium / Q000633:toxicity; D000818:Animals; D002374:Catalase / Q000494:pharmacology; D016923:Cell Death; D002460:Cell Line / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D008054:Lipid Peroxides / Q000032:analysis; D020267:MPTP Poisoning; D009474:Neurons / Q000187:drug effects*; D010293:Pargyline / Q000494:pharmacology; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012642:Selegiline / Q000494:pharmacology; D013482:Superoxide Dismutase / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000378:metabolism
D008054:Lipid Peroxides; D017382:Reactive Oxygen Species; C072770:1-methyl-4-phenyl-1,2,3,6-tetrahydro-3-pyridinol; D012642:Selegiline; D010293:Pargyline; D015632:1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine; D006861:Hydrogen Peroxide; D002374:Catalase; D013482:Superoxide Dismutase; D015655:1-Methyl-4-phenylpyridinium