title: Inhibitory effects of theophylline on the peroxynitrite-augmented release of matrix metalloproteinases by lung fibroblasts.
abstract: The anti-inflammatory effects of theophylline have been reported to include inhibition of the release of proinflammatory mediators from macrophages and neutrophils. Overproduction of reactive nitrogen species (RNS) has been reported in the airways of patients with chronic obstructive pulmonary disease (COPD), and this causes tissue inflammation and injury. We investigated whether peroxynitrite stimulated the release of matrix metalloproteinases 2 and 9 (MMP-2 and -9; gelatinases) from human fetal lung fibroblasts (HFL-1 cell line) and whether theophylline inhibited the peroxynitrite-augmented release of MMPs. HFL-1 cells and primary lung fibroblasts were treated with peroxynitrite (an RNS), and gelatinases levels were evaluated by gelatin zymography. The inhibitory effect of theophylline on the peroxynitrite-augmented release of MMP-2 and MMP-9 was also investigated. To explore the cell signaling pathways involved in the peroxynitrite-induced gelatinases release and the inhibitory effect of theophylline, transforming growth factor-β(1) (TGF-β(1)), nuclear factor-κB (NF-κB), and histone deacetylase (HDAC) were measured. Peroxynitrite significantly augmented the release of MMP-2 and MMP-9 by fibroblasts (P < 0.01), as well as TGF-β(1) release (P < 0.01), NF-κB activation (P < 0.01), and HDAC2 inactivation (P < 0.01). An NF-κB inhibitor diminished the RNS-augmented release of MMPs and TGF-β(1) (P < 0.01), and a neutralizing TGF-β antibody also diminished MMP release (P < 0.01). Theophylline significantly inhibited the peroxynitrite-augmented release of MMP-2 and MMP-9 in HFL-1 cells and normal adult lung fibroblasts, and it also inhibited the peroxynitrite-mediated HDAC2 inactivation, NF-κB activation, and TGF-β(1) release in HFL-1 cells (all P < 0.01). These results suggest that peroxynitrite can influence tissue remodeling by promoting gelatinases release, while theophylline suppresses peroxynitrite-induced tissue remodeling via pathways involving NF-κB/TGF-β(1) and/or HDAC in the HFL-1 cell line.
authors.affiliation: Third Department of Internal Medicine, Wakayama Medical University School of Medicine, Wakayama, Japan. sugiura@wakayama-med.ac.jp
authors.forename: Hisatoshi
authors.initials: H
authors.lastname: Sugiura
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kawabata
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Ichikawa
authors.forename: Akira
authors.initials: A
authors.lastname: Koarai
authors.forename: Satoru
authors.initials: S
authors.lastname: Yanagisawa
authors.forename: Takashi
authors.initials: T
authors.lastname: Kikuchi
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Minakata
authors.forename: Kazuto
authors.initials: K
authors.lastname: Matsunaga
authors.forename: Masanori
authors.initials: M
authors.lastname: Nakanishi
authors.forename: Tsunahiko
authors.initials: T
authors.lastname: Hirano
authors.forename: Keiichiro
authors.initials: K
authors.lastname: Akamatsu
authors.forename: Kanako
authors.initials: K
authors.lastname: Furukawa
authors.forename: Masakazu
authors.initials: M
authors.lastname: Ichinose
mesh_terms: D057134:Antibodies, Neutralizing / Q000008:administration & dosage; D001993:Bronchodilator Agents / Q000494:pharmacology*; D002460:Cell Line; D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology; D006655:Histone Deacetylases / Q000032:analysis; D006801:Humans; D008168:Lung / Q000187:drug effects* / Q000201:enzymology; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D016328:NF-kappa B / Q000032:analysis / Q000037:antagonists & inhibitors; D030421:Peroxynitrous Acid / Q000008:administration & dosage* / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013806:Theophylline / Q000494:pharmacology*; D053773:Transforming Growth Factor beta1 / Q000032:analysis / Q000037:antagonists & inhibitors
chemical_list: D057134:Antibodies, Neutralizing; D001993:Bronchodilator Agents; D016328:NF-kappa B; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1; D030421:Peroxynitrous Acid; D013806:Theophylline; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D006655:Histone Deacetylases