title: Protective effects of carbon monoxide-releasing molecule-2 on the barrier function of intestinal epithelial cells.
abstract: OBJECTIVE
  To investigate the protective effects and mechanisms of carbon monoxide-releasing molecule-2 (CORM-2) on barrier function of intestinal epithelial cells.
  
  
  METHODS
  After pre-incubation with CORM-2 for 1 hour, cultured intestinal epithelial IEC-6 cells were stimulated with 50 µg/ml lipopolysaccharides (LPS). Cytokines levels in culture medium were detected using ELISA kits. Trans-epithelial electrical resistance (TER) of IEC-6 cell monolayers in Transwells were measured with a Millipore electric resistance system (ERS-2; Millipore) and calculated as Ω/cm2 at different time points after LPS treatment. The permeability changes were also measured using FITC-dextran. The levels of tight junction (TJ) proteins (occludin and ZO-1) and myosin light chain (MLC) phosphorylation were detected using Western blotting with specific antibodies. The subsequent structural changes of TJ were visualized using transmission electron microscopy (TEM).
  
  
  RESULTS
  CORM-2 significantly reduced LPS-induced secretion of TNF-α and IL-1β. The LPS-induced decrease of TER and increase of permeability to FITC-dextran were inhibited by CORM-2 in a concentration dependent manner (P<0.05). LPS-induced reduction of tight junction proteins and increase of MLC phosphorylation were also attenuated. In LPS-treated cells, TEM showed diminished electron-dense material and interruption of TJ and desmosomes between the apical lateral margins of adjoining cells, which were prevented by CORM-2 treatment.
  
  
  CONCLUSIONS
  The present study demonstrates that CORM-2, as a novel CO-releasing molecule, has ability to protect the barrier function of LPS-stimulated intestinal epithelial cells. Inhibition of inflammatory cytokines release, restoration of TJ proteins and suppression of MLC phosphorylation are among the protective effects of CORM-2.
authors.affiliation: Department of Critical Care Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjiang, Jiangsu Province, PR China.
authors.forename: Xinwei
authors.initials: X
authors.lastname: Mu
authors.affiliation: Department of Critical Care Medicine, Nanjing Children,s Hospital, Nanjing Medical University, Nanjiang, Jiangsu Province, PR China.
authors.forename: Chen
authors.initials: C
authors.lastname: Pan
authors.affiliation: Department of Critical Care Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjiang, Jiangsu Province, PR China.
authors.forename: Shuyun
authors.initials: S
authors.lastname: Zheng
authors.affiliation: Institute of Infection and Global Health, the University of Liverpool, Liverpool, United Kingdom.
authors.forename: Yasir
authors.initials: Y
authors.lastname: Alhamdi
authors.affiliation: Department of Burn and Plastic Surgery, Affiliated Hospital, Jiangsu University, Zhenjiang, Jiangsu Province, PR China.
authors.forename: Bingwei
authors.initials: B
authors.lastname: Sun
authors.affiliation: Department of Critical Care Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjiang, Jiangsu Province, PR China.
authors.forename: Qiankun
authors.initials: Q
authors.lastname: Shi
authors.affiliation: Department of Critical Care Medicine, Nanjing First Hospital, Nanjing Medical University, Nanjiang, Jiangsu Province, PR China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Burn and Plastic Surgery, Affiliated Hospital, Jiangsu University, Zhenjiang, Jiangsu Province, PR China.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Sun
authors.affiliation: Institute of Infection and Global Health, the University of Liverpool, Liverpool, United Kingdom.
authors.forename: Chenghock
authors.initials: C
authors.lastname: Toh
authors.affiliation: Institute of Infection and Global Health, the University of Liverpool, Liverpool, United Kingdom.
authors.forename: Guozheng
authors.initials: G
authors.lastname: Wang
mesh_terms: D000818:Animals; D002248:Carbon Monoxide / Q000494:pharmacology*; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D046529:Microscopy, Electron, Transmission; D009942:Organometallic Compounds / Q000494:pharmacology*; D020011:Protective Agents / Q000494:pharmacology*; D051381:Rats; D019108:Tight Junctions / Q000187:drug effects / Q000648:ultrastructure
chemical_list: D016207:Cytokines; D008070:Lipopolysaccharides; D009942:Organometallic Compounds; D020011:Protective Agents; C447082:tricarbonyldichlororuthenium (II) dimer; D002248:Carbon Monoxide
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