title: Role of N-acetylcysteine and GSH redox system on total and active MMP-2 in intestinal myofibroblasts of Crohn's disease patients.
abstract: OBJECTIVE
  Intestinal subepithelial myofibroblasts (ISEMFs)(1) are the predominant source of matrix metalloproteinase-2 (MMP-2) in gut, and a decrease in glutathione/oxidized glutathione (GSH/GSSG) ratio, intracellular redox state index, occurs in the ISEMFs of patients with Crohn's disease (CD). The aim of this study is to demonstrate a relationship between MMP-2 secretion and activation and changes of GSH/GSSG ratio in ISEMFs stimulated or not with tumor necrosis factor alpha (TNFα).
  
  
  METHODS
  ISEMFs were isolated from ill and healthy colon mucosa of patients with active CD. Buthionine sulfoximine, GSH synthesis inhibitor, and N-acetylcysteine (NAC), precursor of GSH synthesis, were used to modulate GSH/GSSG ratio. GSH and GSSG were measured by HPLC and MMP-2 by ELISA Kit.
  
  
  RESULTS
  In cells, stimulated or not with TNFα, a significant increase in MMP-2 secretion and activation, related to increased oxidative stress, due to low GSH/GSSG ratio, was detected. NAC treatment, increasing this ratio, reduced MMP-2 secretion and exhibited a direct effect on the secreted MMP-2 activity. In NAC-treated and TNFα-stimulated ISEMFs of CD patients' MMP-2 activity were restored to physiological value. The involvement of c-Jun N-terminal kinase pathway on redox regulation of MMP-2 secretion has been demonstrated.
  
  
  CONCLUSIONS
  For the first time, in CD patient ISEMFs, a redox regulation of MMP-2 secretion and activation related to GSH/GSSG ratio and inflammatory state have been demonstrated. This study suggests that compounds able to maintain GSH/GSSG ratio to physiological values can be useful to restore normal MMP-2 levels reducing in CD patient intestine the dysfunction of epithelial barrier.
authors.affiliation: Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, viale Morgagni 50, 50134 Florence, Italy.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Romagnoli
authors.forename: Tommaso
authors.initials: T
authors.lastname: Marcucci
authors.forename: Lucia
authors.initials: L
authors.lastname: Picariello
authors.forename: Francesco
authors.initials: F
authors.lastname: Tonelli
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Vincenzini
authors.forename: Teresa
authors.initials: T
authors.lastname: Iantomasi
mesh_terms: D000111:Acetylcysteine / Q000378:metabolism*; D000328:Adult; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D003424:Crohn Disease / Q000201:enzymology* / Q000473:pathology*; D004789:Enzyme Activation / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism*; D019803:Glutathione Disulfide / Q000378:metabolism; D006801:Humans; D007422:Intestines / Q000473:pathology*; D042541:Intracellular Space / Q000378:metabolism; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D008875:Middle Aged; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D058628:Myofibroblasts / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology; D010084:Oxidation-Reduction / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D055815:Young Adult
chemical_list: D047428:Protein Kinase Inhibitors; D014409:Tumor Necrosis Factor-alpha; D019328:Buthionine Sulfoximine; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D005978:Glutathione; D019803:Glutathione Disulfide; D000111:Acetylcysteine