Abstract
Matrix metalloproteinase-9 is implicated in airway inflammation and airway remodeling in asthma. We have previously confirmed that human rhinovirus-16 (HRV-16) infection increases MMP-9 expression both in vivo and in vitro. However, the role of the AP-1 sites within the MMP-9 promoter and the effect of commonly used asthma pharmacotherapies in modulating human rhinovirus (HRV)-induced MMP-9 production have not yet been elucidated. Experiments were performed in vitro in the human bronchial epithelial (HBE) cell line BEAS-2B and in primary HBE cells obtained from non-transplanted lungs. Using site-directed mutagenesis approaches, AP-1 sites were found to be necessary for HRV-induced MMP-9 promoter drive. EMSAs and supershift assays identified complexes consisting of Fos-related Ag-1 (Fra-1) in addition to other AP-1 subunits. Small interfering RNA approaches indicated that Fra-1 was induced upon HRV-16 infection in BEAS-2B cells and was necessary for MMP-9 expression in both BEAS-2B and primary HBE cells. Inhibition of MEK1/2 activity using PD98059 and U0126 reduced Fra-1 expression, DNA binding, MMP-9 promoter drive, and MMP-9 protein production. The long-acting β(2)-agonist formoterol and the glucocorticoid dexamethasone significantly reduced HRV-induced ERK phosphorylation, Fra-1, and MMP-9 expression in BEAS-2B cells. These data indicate that HRV-induced activation of the MEK/ERK MAPK pathway and Fra-1 expression are necessary for the upregulation of MMP-9 and can be modulated by two distinct but commonly used asthma pharmacotherapies. Together, these results offer insights into the mechanisms by which long-acting β(2)-agonists and glucocorticoids might reduce HRV-related asthma exacerbations.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Anti-Inflammatory Agents / pharmacology*
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Bronchodilator Agents / pharmacology*
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Butadienes / pharmacology
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Cell Line
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Dexamethasone / pharmacology*
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Enzyme Inhibitors / pharmacology
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Ethanolamines / pharmacology*
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Flavonoids / pharmacology
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Formoterol Fumarate
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Gene Expression Regulation, Enzymologic / drug effects*
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Gene Expression Regulation, Enzymologic / genetics
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Gene Expression Regulation, Enzymologic / immunology
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Humans
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MAP Kinase Kinase 1 / antagonists & inhibitors
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MAP Kinase Kinase 1 / genetics
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MAP Kinase Kinase 1 / immunology
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MAP Kinase Kinase 1 / metabolism
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MAP Kinase Kinase 2 / antagonists & inhibitors
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MAP Kinase Kinase 2 / genetics
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MAP Kinase Kinase 2 / immunology
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MAP Kinase Kinase 2 / metabolism
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MAP Kinase Signaling System / drug effects
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MAP Kinase Signaling System / genetics
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MAP Kinase Signaling System / immunology
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Matrix Metalloproteinase 9 / biosynthesis
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Matrix Metalloproteinase 9 / genetics
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Matrix Metalloproteinase 9 / immunology*
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Nitriles / pharmacology
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Phosphorylation / drug effects
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Phosphorylation / genetics
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Phosphorylation / immunology
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Picornaviridae Infections / drug therapy
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Picornaviridae Infections / enzymology
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Picornaviridae Infections / genetics
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Picornaviridae Infections / immunology*
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Picornaviridae Infections / pathology
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Proto-Oncogene Proteins c-fos / biosynthesis
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Proto-Oncogene Proteins c-fos / genetics
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Proto-Oncogene Proteins c-fos / immunology*
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Response Elements / genetics
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Response Elements / immunology
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Rhinovirus / genetics
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Rhinovirus / immunology*
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Rhinovirus / metabolism
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Transcription Factor AP-1 / genetics
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Transcription Factor AP-1 / immunology
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Transcription Factor AP-1 / metabolism
Substances
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Anti-Inflammatory Agents
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Bronchodilator Agents
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Butadienes
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Enzyme Inhibitors
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Ethanolamines
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Flavonoids
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Nitriles
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Proto-Oncogene Proteins c-fos
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Transcription Factor AP-1
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U 0126
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fos-related antigen 1
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Dexamethasone
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MAP2K2 protein, human
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MAP Kinase Kinase 1
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MAP Kinase Kinase 2
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MAP2K1 protein, human
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Matrix Metalloproteinase 9
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2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one
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Formoterol Fumarate