title: Oxidative state and IL-6 production in intestinal myofibroblasts of Crohn's disease patients.
abstract: BACKGROUND
  Intestinal subepithelial myofibroblasts (ISEMFs) produce inflammatory cytokines in response to certain stimuli. In the intestine of patients with Crohn's disease (CD), cytokine synthesis is modified and an increased number of myofibroblasts has been observed. The intracellular redox state influences cytokine production and oxidative stress is present in the intestinal mucosa of CD patients.
  
  
  METHODS
  This study was performed in ISEMFs isolated from the colon of patients with active CD and in a myofibroblast cell line derived from human colonic mucosa: 18Co cells. Cellular glutathione (GSH) levels were modulated by treatment with buthionine sulfoximine, an inhibitor of GSH synthesis, or N-acetylcysteine, a GSH precursor. GSH and oxidized glutathione (GSSG) levels were measured by high-performance liquid chromatography (HPLC) methods. Interleukin (IL)-6 production was detected by enzyme-linked immunosorbent assay (ELISA).
  
  
  RESULTS
  ISEMFs of CD patients exhibited an increased oxidative state due to a decrease in the GSH/GSSG ratio, which is related to an increase in basal IL-6 production or is stimulated by tumor necrosis factor alpha (TNFα) or bacterial products. This relationship was also confirmed in 18Co cells. Phosphorylation and activation of ERK1/2 and p38 MAPK, which are signaling factors involved in the IL-6 synthesis, were also increased when there is oxidative stress in ISEMFs.
  
  
  CONCLUSIONS
  This study shows for the first time in ISEMFs of CD patients an increased production of IL-6 synthesis related to the decrease in the GSH/GSSH ratio, suggesting redox regulation with the involvement of specific kinase activation. The present data shed light on the pathogenesis of inflammatory chronic processes and relapses that occur in this pathology.
authors: Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Firenze, Italia.
authors: Serena
authors: S
authors: Catarzi
authors: Fabio
authors: F
authors: Favilli
authors: Cecilia
authors: C
authors: Romagnoli
authors: Tommaso
authors: T
authors: Marcucci
authors: Lucia
authors: L
authors: Picariello
authors: Francesco
authors: F
authors: Tonelli
authors: Maria Teresa
authors: MT
authors: Vincenzini
authors: Teresa
authors: T
authors: Iantomasi
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000328:Adult; D019328:Buthionine Sulfoximine / Q000494:pharmacology; D002460:Cell Line; D003106:Colon / Q000378:metabolism / Q000503:physiopathology; D003424:Crohn Disease / Q000378:metabolism* / Q000503:physiopathology; D005260:Female; D005978:Glutathione / Q000187:drug effects / Q000378:metabolism*; D019803:Glutathione Disulfide / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007082:Ileum / Q000378:metabolism / Q000503:physiopathology; D015850:Interleukin-6 / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D058628:Myofibroblasts / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D010766:Phosphorylation / Q000187:drug effects; D015398:Signal Transduction / Q000502:physiology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D015850:Interleukin-6; D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha; D019328:Buthionine Sulfoximine; D009240:N-Formylmethionine Leucyl-Phenylalanine; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases; D005978:Glutathione; D019803:Glutathione Disulfide; D000111:Acetylcysteine