title: Protective effect of hydrogen sulfide on TNF-α and IFN-γ-induced injury of intestinal epithelial barrier function in Caco-2 monolayers.
abstract: OBJECTIVE
  Studies have verified the protective effect of Hydrogen Sulfide (H2S) on gastric ulcer and ulcerative colitis, but the mechanisms are not fully illustrated. In this study, the possible protective effect of H2S on TNF-α/IFN-γ induced barrier dysfunction was investigated in Caco-2 cell monolayers.
  
  
  METHODS
  The barrier function of Caco-2 monolayers was evaluated by measuring trans-epithelial electrical resistance (TEER) and FITC-Dextran 4 kDa (FD-4) trans-membrane flux. ZO-1 and Occludin were chosen as markers of the localization of tight junction (TJ) proteins for immunofluorescence. The expression of MLCK and phosphorylation level of myosin light chain (MLC) were measured by immunoblotting. The activation of NF-kB p65 was analyzed by EMSA and immunofluorescence.
  
  
  RESULTS
  NaHS at 500 uM significantly attenuated TNF-α/IFN-γ-indueced Caco-2 monolayer barrier injury. The increased expression of MLCK and increased phosphorylation level of MLC induced by TNF-α/IFN-γ was also inhibited significantly by NaHS. Additionally, NaHS inhibited TNF-α/IFN-γ induced activation and nuclear translocation of NF-kB p65.
  
  
  CONCLUSIONS
  The present study reveals the protective effect of H2S on TNF-α and IFN-γ-induced injury of intestinal epithelial barrier function in Caco-2 monolayers and suggests that the suppression of MLCK-P-MLC signaling mediated by NF-kB P65 might be one of the mechanisms underlying the protective effect of H2S.
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China.
authors.forename: Shan-Wen
authors.initials: SW
authors.lastname: Chen
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Zuo
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China.
authors.forename: Jun-Ling
authors.initials: JL
authors.lastname: Zhang
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China.
authors.forename: Zi-Yi
authors.initials: ZY
authors.lastname: Chen
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China.
authors.forename: Guo-Wei
authors.initials: GW
authors.lastname: Chen
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China.
authors.forename: Yi-Sheng
authors.initials: YS
authors.lastname: Pan
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China.
authors.forename: Yu-Cun
authors.initials: YC
authors.lastname: Liu
authors.affiliation: Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People's Republic of China. wangpengyuan2014@126.com.
authors.forename: Peng-Yuan
authors.initials: PY
authors.lastname: Wang
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D018938:Caco-2 Cells; D004848:Epithelium / Q000187:drug effects*; D006801:Humans; D006862:Hydrogen Sulfide / Q000494:pharmacology*; D007371:Interferon-gamma / Q000037:antagonists & inhibitors* / Q000633:toxicity*; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects*; D018994:Myosin Light Chains / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D062793:Occludin / Q000378:metabolism; D010766:Phosphorylation; D013276:Stomach Ulcer / Q000473:pathology / Q000517:prevention & control; D062725:Tight Junction Proteins / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000633:toxicity*
chemical_list: D015415:Biomarkers; D018994:Myosin Light Chains; D062793:Occludin; D062725:Tight Junction Proteins; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D009219:Myosin-Light-Chain Kinase; D006862:Hydrogen Sulfide
keywords: H2S; MLCK; NF-kB; Tight junction; ZO-1