OPC-compounds prevent oxidant-induced carbonylation and depolymerization of the F-actin cytoskeleton and intestinal barrier hyperpermeability.
Rebamipide (OPC-12759), a quinolone derivative, and OPC-6535, a thiazol-carboxylic acid derivative, are compounds with ability to protect gastrointestinal (GI) mucosal integrity against reactive oxygen metabolites (ROM). The underlying mechanism of OPC-mediated protection remains poorly understood. It is now established that ROM can injure the mucosa by disruption of the cytoskeletal network, a key component of mucosal barrier integrity. We, therefore, investigated whether OPC compounds prevent the oxidation, disassembly, and instability of the cytoskeletal protein actin and, in turn, protect intestinal barrier function against ROM. Human intestinal (Caco-2) cell monolayers were pretreated with OPC (-12759 or -6535) prior to incubation with ROM (H2O2) or HOCl). Effects on cell integrity (ethidium homodimer-1), epithelial barrier function (fluorescein sulfonic acid clearance), and actin cytoskeletal integrity (high-resolution laser confocal) were then determined. Cells were also processed for quantitative immunoblotting of G- and F-actin to measure oxidation (carbonylation) and disassembly of actin. In monolayers exposed to ROM, preincubation with OPC compounds prevented actin oxidation, decreased depolymerized G-actin, and enhanced the stable F-actin. Concomitantly, OPC agents abolished both actin cytoskeletal disruption and monolayer barrier dysfunction. Data suggest for the first time that OPC drugs prevent oxidation of actin and lead to the protection of actin cytoskeleton and intestinal barrier integrity against oxidant insult. Accordingly, these compounds may be used as novel therapeutic agents for the treatment of a variety of oxidative inflammatory intestinal disorders with an abnormal mucosal barrier such as inflammatory bowel disease.
Banan
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Departments of Internal Medicine (Division of Digestive Diseases), Pharmacology, and Molecular Biophysics and Physiology, Rush University Medical Center, 1725 W. Harrison, Chicago, IL 60612, USA. ali_banan@rush.edu
Fitzpatrick
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Zhang
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Y
Keshavarzian
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A
D000199:Actins / Q000032:analysis / Q000378:metabolism*; D000409:Alanine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology*; D015153:Blotting, Western; D018938:Caco-2 Cells; D003599:Cytoskeleton / Q000378:metabolism*; D005456:Fluorescent Dyes; D006801:Humans; D015151:Immunoblotting; D007413:Intestinal Mucosa / Q000187:drug effects*; D008856:Microscopy, Fluorescence; D016877:Oxidants / Q000494:pharmacology*; D010084:Oxidation-Reduction; D010539:Permeability; D015363:Quinolones / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology*
D000199:Actins; D000975:Antioxidants; D005456:Fluorescent Dyes; D016877:Oxidants; D015363:Quinolones; D017382:Reactive Oxygen Species; D013844:Thiazoles; C052785:rebamipide; D000409:Alanine; C108965:tetomilast