title: Release of Matrix Metalloproteinases-2 and 9 by S-Nitrosylated Caveolin-1 Contributes to Degradation of Extracellular Matrix in tPA-Treated Hypoxic Endothelial Cells.
abstract: Intracranial hemorrhage remains the most feared complication in tissue plasminogen activator (tPA) thrombolysis for ischemic stroke. However, the underlying molecular mechanisms are still poorly elucidated. In this study, we reported an important role of caveolin-1 (Cav-1) s-nitrosylation in matrix metalloproteinase (MMP)-2 and 9 secretion from tPA-treated ischemic endothelial cells. Brain vascular endothelial cells (bEND3) were exposed to oxygen-glucose deprivation (OGD) for 2 h before adding recombinant human tPA for 6 h. This treatment induced a significant increase of MMP2 and 9 in the media of bEND3 cells and a simultaneous degradation of fibronectin and laminin β-1, the two main components of extracellular matrix (ECM). Inhibition of MMP2 and 9 with SB-3CT completely blocked the degradation of fibronectin and laminin β-1. ODG+tPA treatment led to Cav-1 shedding from bEND3 cells into the media. Notably, OGD triggered nitric oxide (NO) production and S-nitrosylationof Cav-1 (SNCav-1). Meanwhile tPA induced activation of ERK signal pathway and stimulates the secretion of SNCav-1. Pretreatment of bEND3 cells with C-PTIO (a NO scavenger) or U0126 (a specific ERK inhibitor) significantly reduced OGD-induced S-nitrosylation of Cav-1 in cells and blocked the secretion of Cav-1 and MMP2 and 9 into the media as well as the degradation of fibronectin and laminin β-1 in OGD and tPA-treated cells. These data indicate that OGD-triggered Cav-1 S-nitrosylation interacts with tPA-induced ERK activation to augment MMP2 and 9 secretion and subsequent ECM degradation, which may account for the exacerbation of ischemic blood brain barrier damage following tPA thrombolysis for ischemic stroke.
authors.affiliation: Department of Cardiology, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Haoming
authors.initials: H
authors.lastname: Song
authors.affiliation: Department of Neurology, Guangzhou Brain Hospital, Affiliated Hospital of Guangzhou Medical University, Guangzhou 510370, China.
authors.forename: Youjun
authors.initials: Y
authors.lastname: Cheng
authors.affiliation: Translational Center for Stem Cell Research, Tongji Hospital, Stem Cell Research Center, Tongji University School of Medicine, Shanghai 200065, China.|Department of orthopedics, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Bi
authors.affiliation: Translational Center for Stem Cell Research, Tongji Hospital, Stem Cell Research Center, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Yihui
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Translational Center for Stem Cell Research, Tongji Hospital, Stem Cell Research Center, Tongji University School of Medicine, Shanghai 200065, China.|Department of general surgery, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Jun
authors.affiliation: Department of Cardiology, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China.|Translational Center for Stem Cell Research, Tongji Hospital, Stem Cell Research Center, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Wenlin
authors.initials: W
authors.lastname: Ma
authors.affiliation: Translational Center for Stem Cell Research, Tongji Hospital, Stem Cell Research Center, Tongji University School of Medicine, Shanghai 200065, China.|Department of general surgery, Tongji Hospital, Tongji University School of Medicine, Shanghai 200065, China.
authors.forename: Huimin
authors.initials: H
authors.lastname: Wu
mesh_terms: D000818:Animals; D001565:Benzoates / Q000494:pharmacology; D051242:Caveolin 1 / Q000378:metabolism*; D015687:Cell Hypoxia / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D005109:Extracellular Matrix / Q000378:metabolism*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D005947:Glucose / Q000172:deficiency; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007797:Laminin / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D015538:Nitrosation; D010100:Oxygen / Q000378:metabolism; D059748:Proteolysis / Q000187:drug effects; D010959:Tissue Plasminogen Activator / Q000008:administration & dosage / Q000494:pharmacology*
chemical_list: D001565:Benzoates; D051242:Caveolin 1; D017077:Culture Media, Conditioned; D005353:Fibronectins; D007093:Imidazoles; D007797:Laminin; C079393:1,3-dihydroxy-4,4,5,5-tetramethyl-2-(4-carboxyphenyl)tetrahydroimidazole; D009569:Nitric Oxide; D048049:Extracellular Signal-Regulated MAP Kinases; D010959:Tissue Plasminogen Activator; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D005947:Glucose; D010100:Oxygen