| Pharmacological inhibition of neuronal NADPH oxidase protects against 1-methyl-4-phenylpyridinium (MPP+)-induced oxidative stress and apoptosis in mesencephalic dopaminergic neuronal cells. | |
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MedLine Citation:
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PMID: 17904225 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Oxidative stress is widely recognized as a key mediator of degenerative processes in Parkinson's disease (PD). Recently, we demonstrated that the dopaminergic toxin MPP+ initiates oxidative stress to cause caspase-3-dependent apoptotic cell death in mesencephalic dopaminergic neuronal (N27) cells. In this study, we determined the source of reactive oxygen species (ROS) produced during MPP+-induced apoptotic cell death. In addition to mitochondria, plasma membrane NADPH oxidase is considered a major producer of ROS inside the cell. Here, we show that N27 neuronal cells express key NADPH oxidase subunits gp91phox and p67phox. We used structurally diverse NADPH oxidase inhibitors, aminoethyl-benzenesulfonylfluoride (AEBSF, 100-1000microM), apocynin (100-1000microM), and diphenylene iodonium (DPI, 3-30microM), to inhibit intrinsic NADPH oxidase activity in N27 cells. Flow cytometric analysis using the ROS-sensitive dye hydroethidine revealed that AEBSF blocked 300microM MPP+-induced ROS production for over 45min in N27 cells, in a dose-dependent manner. Further treatment with DPI, apocynin, and SOD also blocked MPP+-induced ROS production. In Sytox cell death assays, co-treatment with AEBSF, apocynin, or DPI for 24h significantly suppressed MPP+-induced cytotoxic cell death. Similarly, co-treatment with these inhibitors also significantly attenuated MPP+-induced increases in caspase-3 enzymatic activity. Furthermore, quantitative DNA fragmentation ELISA assays revealed that AEBSF, DPI, and apocynin rescue N27 cells from MPP+-induced apoptotic cell death. Together, these results indicate for the first time that intracellular ROS generated by NAPDH oxidase are present within the mesencephalic neuronal cells, and are a key determinant of MPP+-mediated dopaminergic degeneration in in vitro models of dopaminergic degeneration. This study supports a critical role of NADPH oxidase in the oxidative damage in PD; targeting this enzyme may lead to novel therapies for PD. |
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Authors:
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Vellareddy Anantharam; Siddharth Kaul; Chunjuan Song; Arthi Kanthasamy; Anumantha G Kanthasamy |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2007-08-25 |
Journal Detail:
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Title: Neurotoxicology Volume: 28 ISSN: 0161-813X ISO Abbreviation: Neurotoxicology Publication Date: 2007 Sep |
Date Detail:
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Created Date: 2007-10-09 Completed Date: 2007-12-26 Revised Date: 2011-04-26 |
Medline Journal Info:
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Nlm Unique ID: 7905589 Medline TA: Neurotoxicology Country: Netherlands |
Other Details:
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Languages: eng Pagination: 988-97 Citation Subset: IM |
Affiliation:
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Parkinson's Disorder Research Laboratory, Iowa Center for Advanced Neurotoxicology, Department of Biomedical Sciences, Iowa State University, Ames, IA 50011-1250, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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1-Methyl-4-phenylpyridinium
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antagonists & inhibitors*,
toxicity* Animals Apoptosis / drug effects* Blotting, Western Caspases / metabolism Cell Survival / drug effects DNA Fragmentation / drug effects Dopamine / physiology* Dopamine Agents / toxicity* Dose-Response Relationship, Drug Enzyme Inhibitors / pharmacology* MPTP Poisoning / metabolism*, prevention & control* Membrane Glycoproteins / genetics Mesencephalon / cytology, drug effects* NADPH Oxidase / antagonists & inhibitors*, genetics Neurons / drug effects, enzymology* Organic Chemicals Oxidative Stress / drug effects* Phosphoproteins / genetics Rats Reactive Oxygen Species / metabolism |
| Grant Support | |
ID/Acronym/Agency:
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ES10586/ES/NIEHS NIH HHS; NS38644/NS/NINDS NIH HHS; NS45133/NS/NINDS NIH HHS; R01 ES010586-07/ES/NIEHS NIH HHS; R01 NS038644-09/NS/NINDS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Dopamine Agents; 0/Enzyme Inhibitors; 0/Membrane Glycoproteins; 0/Organic Chemicals; 0/Phosphoproteins; 0/Reactive Oxygen Species; 0/SYTOX Green; 0/neutrophil cytosol factor 67K; 48134-75-4/1-Methyl-4-phenylpyridinium; EC 1.6.-/Cybb protein, rat; EC 1.6.3.1/NADPH Oxidase; EC 3.4.22.-/Caspases |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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