title: A leading role for NADPH oxidase in an in-vitro study of experimental autoimmune encephalomyelitis.
abstract: Myelin oligodendrocyte glycoprotein peptide fragment 35-55 (MOG35-55) is a major autoantigen inducing experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis that is characterized by blood-brain barrier (BBB) disruption. Various experimental approaches have employed MOG35-55 in vivo; however, in vitro BBB models using MOG35-55 are rarely reported. We investigated MOG35-55 exposure effects with complete Freund's adjuvant (CFA) and pertussis toxin (PTX) on brain endothelial cells and elucidated the relationships among NADPH oxidase, MMP-9, ICAM-1, and VCAM-1. These 4 factors significantly increased in MOG35-55+CFA+PTX-exposed endothelial cells compared with the control cells. NADPH oxidase inhibition using apocynin reduced MMP-9 activity, ICAM-1, and VCAM-1. MMP-9 inhibitor I decreased expression of ICAM-1 and VCAM-1, and both anti-ICAM-1 and anti-VCAM-1 inhibited MMP-9 activity. Inhibitions of MMP-9, ICAM-1, and VCAM-1 did not change NADPH oxidase activity. Although inhibition of these 4 factors decreased BBB permeability in cells, inhibition of NADPH oxidase exhibited the highest decrease among these. NADPH oxidase directly influenced MMP-9, ICAM-1, and VCAM-1, but not vice versa. MMP-9 and the cell adhesion molecules reversibly affected each other. In conclusion, NADPH oxidase-derived superoxide elevated expression of MMP-9, ICAM-1, and VCAM-1, and these interactions can finally result in increases of BBB permeability in MOG35-55+CFA+PTX-exposed endothelial cells.
authors.affiliation: Toxicology Laboratory, Doping Control Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology, Daejeon 34113, Republic of Korea.
authors.forename: Ji-Eun
authors.initials: JE
authors.lastname: Seo
authors.affiliation: Toxicology Laboratory, Doping Control Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology, Daejeon 34113, Republic of Korea.
authors.forename: Mahbub
authors.initials: M
authors.lastname: Hasan
authors.affiliation: Toxicology Laboratory, Doping Control Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology, Daejeon 34113, Republic of Korea.
authors.forename: Khandoker Asiqur
authors.initials: KA
authors.lastname: Rahaman
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology, Daejeon 34113, Republic of Korea.
authors.forename: Min-Jung
authors.initials: MJ
authors.lastname: Kang
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology, Daejeon 34113, Republic of Korea.
authors.forename: Byung-Hwa
authors.initials: BH
authors.lastname: Jung
authors.affiliation: Toxicology Laboratory, Doping Control Center, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea; Department of Biological Chemistry, Korea University of Science and Technology, Daejeon 34113, Republic of Korea. Electronic address: oskwon@kist.re.kr.
authors.forename: Oh-Seung
authors.initials: OS
authors.lastname: Kwon
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000378:metabolism*; D002478:Cells, Cultured; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000201:enzymology*; D042783:Endothelial Cells / Q000378:metabolism; D005620:Freund's Adjuvant; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D063308:Myelin-Oligodendrocyte Glycoprotein; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010446:Peptide Fragments; D037342:Pertussis Toxin; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: C090509:Icam1 protein, mouse; D063308:Myelin-Oligodendrocyte Glycoprotein; D010446:Peptide Fragments; D019010:Vascular Cell Adhesion Molecule-1; C445081:myelin oligodendrocyte glycoprotein (35-55); D018799:Intercellular Adhesion Molecule-1; D005620:Freund's Adjuvant; D019255:NADPH Oxidases; D037342:Pertussis Toxin; D020780:Matrix Metalloproteinase 9; C501830:Mmp9 protein, mouse
keywords: ICAM-1; MMP-9; MOG(35–55)+CFA+PTX-exposed endothelial cells; NADPH oxidase; VCAM-1