title: Methamphetamine alters occludin expression via NADPH oxidase-induced oxidative insult and intact caveolae.
abstract: Methamphetamine (METH) is a drug of abuse with neurotoxic and vascular effects that may be mediated by reactive oxygen species (ROS). However, potential sources of METH-induced generation of ROS are not fully understood. This study is focused on the role of NAD(P)H oxidase (NOX) in METH-induced dysfunction of brain endothelial cells. Treatment with METH induced a time-dependent increase in phosphorylation of NOX subunit p47, followed by its binding with gp91 and p22, and the formation of an active NOX complex. An increase in NOX activity was associated with elevated production of ROS, alterations of occludin levels and increased transendothelial migration of monocytes. Inhibition of NOX by NSC 23766 attenuated METH-induced ROS generation, changes in occludin protein levels and monocyte migration. Because an active NOX complex is localized to caveolae, we next evaluated the role of caveolae in METH-mediated toxicity to brain endothelial cells. Treatment with METH induced phosphorylation of ERK1/2 and caveolin-1 protein. Inhibition of ERK1/2 activity or caveolin-1 silencing protected against METH-induced alterations of occludin levels. These findings indicate an important role of NOX and functional caveolae in METH-induced oxidative stress in brain endothelial cells that contribute to the subsequent alterations of occludin levels and transendothelial migration of inflammatory cells.
authors: Department of Neurosurgery, University of Kentucky, KY 40536, USA.
authors: Minseon
authors: M
authors: Park
authors: Bernhard
authors: B
authors: Hennig
authors: Michal
authors: M
authors: Toborek
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism / Q000473:pathology*; D021941:Caveolae / Q000378:metabolism*; D051242:Caveolin 1 / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement; D003573:Cytochrome b Group / Q000037:antagonists & inhibitors / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D008565:Membrane Proteins / Q000378:metabolism*; D008694:Methamphetamine / Q000378:metabolism* / Q000494:pharmacology / Q000633:toxicity; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism; D062793:Occludin; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D017382:Reactive Oxygen Species; D011971:Receptors, Immunologic / Q000378:metabolism
chemical_list: D051242:Caveolin 1; D003573:Cytochrome b Group; D008565:Membrane Proteins; D062793:Occludin; C568986:Ocln protein, mouse; C105949:Pirb protein, mouse; D017382:Reactive Oxygen Species; D011971:Receptors, Immunologic; D008694:Methamphetamine; D019255:NADPH Oxidases; C515614:Cyba protein, mouse; D020928:Mitogen-Activated Protein Kinases