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Inflammation Research

, Volume 64, Issue 10, pp 789–797 | Cite as

Protective effect of hydrogen sulfide on TNF-α and IFN-γ-induced injury of intestinal epithelial barrier function in Caco-2 monolayers

  • Shan-wen Chen
  • Jing Zhu
  • Shuai Zuo
  • Jun-ling Zhang
  • Zi-yi Chen
  • Guo-wei Chen
  • Xin Wang
  • Yi-sheng Pan
  • Yu-cun Liu
  • Peng-yuan Wang
Original Research Paper

Abstract

Background and aim

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.

Method

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 500uM 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.

Conclusion

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.

Keywords

H2NF-kB Tight junction ZO-1 MLCK 

Notes

Acknowledgments

We thank Professor Dingfang Pu for his excellent technical instructions in the measurement of Caco-2 monolayer permeability.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this paper.

Funding

Funding for this work came from the China Health and Medical Development Foundation (Grant: Research on Surgical Infection Mechanism and Control).

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Copyright information

© Springer Basel 2015

Authors and Affiliations

  • Shan-wen Chen
    • 1
  • Jing Zhu
    • 1
  • Shuai Zuo
    • 1
  • Jun-ling Zhang
    • 1
  • Zi-yi Chen
    • 1
  • Guo-wei Chen
    • 1
  • Xin Wang
    • 1
  • Yi-sheng Pan
    • 1
  • Yu-cun Liu
    • 1
  • Peng-yuan Wang
    • 1
  1. 1.Division of General Surgery, Peking University First HospitalPeking UniversityBeijingPeople’s Republic of China

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