title: Redox regulation of MMP-3/TIMP-1 ratio in intestinal myofibroblasts: effect of N-acetylcysteine and curcumin.
abstract: Matrix metalloproteinases (MMPs) play a critical role in inflammation and ulcerations in gut of Crohn׳s disease (CD) patients. Intestinal subepithelial myofibroblasts (ISEMFs) secrete MMPs in response to inflammatory stimuli. Previous data showed in CD-ISEMFs increased oxidative status. The aim of this study was to investigate the role of ISEMFs in modulating the production of MMP-3 and TIMP-1, an inhibitor of MMPs activity. A relationship among oxidative stress, activity of antioxidants and MMP-3/TIMP-1 was also studied. ISEMFs isolated from CD patient colon and human colonic cell line of myofibroblasts (18Co) were used. Oxidative state was modulated by buthionine sulfoximine, an inhibitor of glutathione (GSH) synthesis, and N-acetylcysteine (NAC), GSH precursor. An up-regulation of MMP-3 due to increased oxidative state was found in CD-ISEMFs. Stimulation by tumor necrosis factor (TNF)α increased further MMP-3 levels. On the contrary, no change in TIMP-1 production was determined. NAC treatment decreased MMP-3 production in CD-ISMEFs and removed the enhancement due to TNFα. Similar effects were observed in 18Co cells treated with curcumin, antioxidant with anti-inflammatory properties. The involvement of MAPKs on MMP-3 redox regulation was also shown. This study demonstrates the involvement of ISEMFs and high oxidative state in the increased MMP-3 production found in intestinal mucosa of CD patients. NAC and curcumin normalize MMP-3 levels mainly in TNFα stimulated cells. A modulation of MMP-3 production by NAC and curcumin due to their direct action on transcriptional factors has been also suggested. Therefore, they could have a therapeutic use for the prevention and treatment of fistulaes in CD.
authors.affiliation: Dipartimento di Scienze Biomediche Sperimentali e Cliniche (Sezione Biochimica), Università degli Studi di Firenze, viale Morgagni 50, 50134 Firenze, Italia.
authors.forename: Filippo
authors.initials: F
authors.lastname: Fontani
authors.affiliation: Dipartimento di Chirurgia e Medicina Traslazionale (DCMT) Largo Brambilla 3, 50134 Firenze, Italia.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Marcucci
authors.affiliation: Dipartimento di Scienze Biomediche Sperimentali e Cliniche (Sezione Cubo) viale Morgagni 85, 50134 Firenze, Italia.
authors.forename: Lucia
authors.initials: L
authors.lastname: Picariello
authors.affiliation: Dipartimento di Chirurgia e Medicina Traslazionale (DCMT) Largo Brambilla 3, 50134 Firenze, Italia.
authors.forename: Francesco
authors.initials: F
authors.lastname: Tonelli
authors.affiliation: Dipartimento di Scienze Biomediche Sperimentali e Cliniche (Sezione Biochimica), Università degli Studi di Firenze, viale Morgagni 50, 50134 Firenze, Italia.
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Vincenzini
authors.affiliation: Dipartimento di Scienze Biomediche Sperimentali e Cliniche (Sezione Biochimica), Università degli Studi di Firenze, viale Morgagni 50, 50134 Firenze, Italia. Electronic address: tiantomasi@unifi.it.
authors.forename: Teresa
authors.initials: T
authors.lastname: Iantomasi
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D000328:Adult; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002478:Cells, Cultured; D003106:Colon / Q000166:cytology / Q000378:metabolism; D003424:Crohn Disease / Q000188:drug therapy / Q000201:enzymology*; D003474:Curcumin / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016166:Free Radical Scavengers / Q000494:pharmacology; D005978:Glutathione / Q000096:biosynthesis; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007413:Intestinal Mucosa / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis / Q000378:metabolism*; D008875:Middle Aged; D058628:Myofibroblasts / Q000166:cytology / Q000378:metabolism*; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D015854:Up-Regulation; D055815:Young Adult
chemical_list: D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D014409:Tumor Necrosis Factor-alpha; D019328:Buthionine Sulfoximine; D006861:Hydrogen Peroxide; D019278:Matrix Metalloproteinase 3; D005978:Glutathione; D003474:Curcumin; D000111:Acetylcysteine
keywords: Crohn׳s disease; Curcumin; MMP-3; N-acetylcysteine; Oxidative state