title: Membrane lipid peroxidation in neurodegeneration: Role of thrombin and proteinase-activated receptor-1.
abstract: Thrombin and membrane lipid peroxidation (MLP) have been implicated in various central nervous system (CNS) disorders from CNS trauma to stroke, Alzheimer's (AD) and Parkinson's (PD) diseases. Because thrombin also induces MLP in platelets and its involvement in neurodegenerative diseases we hypothesized that its deleterious effects might, in part, involve formation of MLP in neuronal cells. We previously showed that thrombin induced caspase-3 mediated apoptosis in motor neurons, via a proteinase-activated receptor (PAR1). We have now investigated thrombin's influence on the oxidative state of neurons leading to induction of MLP-protein adducts. Translational relevance of thrombin-induced MLP is supported by increased levels of 4-hydroxynonenal-protein adducts (HNEPA) in AD and PD brains. We now report for the first time that thrombin dose-dependently induces formation of HNEPA in NSC34 mouse motor neuron cells using anti-HNE and anti-acrolein monoclonal antibodies. The most prominent immunoreactive band, in SDS-PAGE, was at ∼54kDa. Membrane fractions displayed higher amounts of the protein-adduct than cytosolic fractions. Thrombin induced MLP was mediated, at least in part, through PAR1 since a PAR1 active peptide, PAR1AP, also elevated HNEPA levels. Of interest, glutamate and Fe2SO4 also increased the ∼54kDa HNEPA band in these cells but to a lesser extent. Taken together our results implicate the involvement of thrombin and MLP in neuronal cell loss observed in various CNS degenerative and traumatic pathologies.
authors.affiliation: Neurobiology Research Laboratory, Department of Veterans Affairs Medical Center, Kansas City, Missouri 64128; Departments of Neurology, University of Kansas Medical Center, Kansas City, Kansas 66170.
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Citron
authors.affiliation: Neurobiology Research Laboratory, Department of Veterans Affairs Medical Center, Kansas City, Missouri 64128.
authors.forename: Syed
authors.initials: S
authors.lastname: Ameenuddin
authors.affiliation: Laboratory of Food and Biodynamics, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Uchida
authors.affiliation: Neurobiology Research Laboratory, Department of Veterans Affairs Medical Center, Kansas City, Missouri 64128; Departments of Neurology, University of Kansas Medical Center, Kansas City, Kansas 66170.
authors.forename: William Z
authors.initials: WZ
authors.lastname: Suo
authors.affiliation: Neurobiology Research Laboratory, Department of Veterans Affairs Medical Center, Kansas City, Missouri 64128; Departments of Pathology and Laboratory Medicine University of Kansas Medical Center, Kansas City, Kansas 66170.
authors.forename: Karen
authors.initials: K
authors.lastname: SantaCruz
authors.affiliation: Neurobiology Research Laboratory, Department of Veterans Affairs Medical Center, Kansas City, Missouri 64128; Departments of Neurology, University of Kansas Medical Center, Kansas City, Kansas 66170; Departments of Pharmacology, Toxicology and Therapeutics, University of Kansas Medical Center, Kansas City, Kansas 66170; Departments of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, Kansas 66170. Electronic address: bwfestoff@mac.com.
authors.forename: Barry W
authors.initials: BW
authors.lastname: Festoff
mesh_terms: D000171:Acrolein / Q000378:metabolism; D000447:Aldehydes / Q000378:metabolism; D000544:Alzheimer Disease / Q000201:enzymology / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D008563:Membrane Lipids / Q000378:metabolism*; D051379:Mice; D009046:Motor Neurons / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D010300:Parkinson Disease / Q000201:enzymology / Q000378:metabolism*; D044463:Receptor, PAR-1 / Q000819:agonists / Q000378:metabolism*; D013917:Thrombin / Q000008:administration & dosage*
chemical_list: D000447:Aldehydes; D008563:Membrane Lipids; D044463:Receptor, PAR-1; D000171:Acrolein; D013917:Thrombin; C027576:4-hydroxy-2-nonenal
keywords: Apoptosis; Membrane lipid peroxidation; Neurodegenerative diseases; Reactive oxygen; Spinal cord injury