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

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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.

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References

  1. Turner JR. Intestinal mucosal barrier function in health and disease. Nat Rev Immunol. 2009;9(11):799–809.

    Article  CAS  PubMed  Google Scholar 

  2. Lin N, Xu LF, Sun M. The protective effect of trefoil factor 3 on the intestinal tight junction barrier is mediated by toll-like receptor 2 via a PI3 K/Akt dependent mechanism. Biochem Biophys Res Commun. 2013;440(1):143–9.

    Article  CAS  PubMed  Google Scholar 

  3. Chen SW, Wang PY, Zhu J, Chen GW, Zhang JL, Chen ZY, Zuo S, Liu YC, Pan YS. Protective effect of 1,25-Dihydroxyvitamin D3 on lipopolysaccharide-induced intestinal epithelial tight Junction injury in Caco-2 cell monolayers. Inflammation. 2014;38:375e383.

    Google Scholar 

  4. Chen S, Zhu J, Chen G, Zuo S, Zhang J, Chen Z, et al. 1,25-Dihydroxyvitamin D3 preserves intestinal epithelial barrier function from TNF-α induced injury via suppression of NF-kB p65 mediated MLCK-P-MLC signaling pathway. Biochem Biophys Res Commun. 2015;. doi:10.1016/j.bbrc.2015.03.125 (Epub ahead of print).

    Google Scholar 

  5. Zanardo RC, Brancaleone V, Distrutti E, et al. Hydrogen sulfide is an endogenous modulator of leukocyte-mediated inflammation. FASEB J. 2006;20:2118–20.

    Article  CAS  PubMed  Google Scholar 

  6. Du J, Huang Y, Yan H, Zhang Q, Zhao M, et al. Hydrogen sulfide suppresses oxidized low-density lipoprotein (ox-LDL)-stimulated monocyte chemoattractant protein 1 generation from macrophages via the nuclear factor κB (NF-κB) pathway. J Biol Chem. 2014;289(14):9741–53.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Wallace JL, Vong L, McKnight W, Dicay M, Martin GR. Endogenous and exogenous hydrogen sulfide promotes resolution of colitis in rats. Gastroenterology. 2009;137(2):569–78.

    Article  CAS  PubMed  Google Scholar 

  8. Flannigan KL, Agbor TA, Blackler RW, Kim JJ, Khan WI, Verdu EF, et al. Impaired hydrogen sulfide synthesis and IL-10 signaling underlie hyperhomocysteinemia-associated exacerbation of colitis. Proc Natl Acad Sci U S A. 2014;111(37):13559–64.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Medani M, Collins D, Docherty NG, Baird AW, O’Connell PR, Winter DC. Emerging role of hydrogen sulfide in colonic physiology and pathophysiology. Inflamm Bowel Dis. 2011;17(7):1620–5.

    Article  PubMed  Google Scholar 

  10. Ye D, Guo S, Al-Sadi R, Ma TY. MicroRNA regulation of intestinal epithelial tight junction permeability. Gastroenterology. 2011;141(4):1323–33.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Puthia MK, Sio SW, Lu J, Tan KS. Blastocystis ratti induces contact- independent apoptosis, F-actin rearrangement, and barrier function disruption in IEC-6 cells. Infect Immun. 2006;74:4114.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Ma TY, Hoa NT, Tran DD, Bui V, Pedram A, Mills S, Merryfield M. Cyto- chalasin B modulation of Caco-2 tight junction barrier: role of myosin light chain kinase. Am. J. Physiol. Gastrointest. Liver Physiol. 2000;279:875e885.

    Google Scholar 

  13. Ma TY, Nguyen D, Bui V, et al. Ethanol modulation of intestinal epithelial tight junction barrier. Am J Physiol Gastrointest Liver Physiol. 1999;276:965e974.

    Google Scholar 

  14. Guo S, Al-Sadi R, Said HM, Ma TY. Lipopolysaccharide causes an increase in intestinal tight junction permeability in vitro and in vivo by inducing enter- ocyte membrane expression and localization of TLR-4 and CD14. Am. J. Pathol. 2013;182:375.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  15. Schlegel N, Meir M, Spindler V, Germer CT, Waschke J. Differential role of RhoGTPases in intestinal epithelial barrier regulation in vitro. J Cell. Physi- ology. 2011;226:1196e1203.

    Google Scholar 

  16. Schliwa M. Action of cytochalasin D on cytoskeletal networks. J Cell Biol. 1982;92:79e91.

    Article  Google Scholar 

  17. Siddiqui MR, Mayanil CS, Kim KS, Tomita T. Angiopoietin-1 Regulates Brain Endothelial Permeability through PTPN-2 Mediated Tyrosine Dephosphorylation of Occludin. PLoS ONE. 2015;10(6):e0130857.

    Article  PubMed Central  PubMed  Google Scholar 

  18. Qasim M, Rahman H, Ahmed R, Oellerich M, Asif AR. Mycophenolic acid mediated disruption of the intestinal epithelial tight junctions Exp. Cell. Res. 2014;322:277e289.

    Article  Google Scholar 

  19. Shen L, Black ED, Witkowski ED, Lencer WI, Guerriero V, Schneeberger EE, Turner JR. Myosin light chain phosphorylation regulates barrier function by remodeling tight junction structure. J. Cell. Sci. 2006;119:2095e2106.

    Article  Google Scholar 

  20. Al-Sadi R, Ye D, Dokladny K, Ma TY. Mechanism of IL-1beta induced increase in intestinal epithelial tight junction permeability. J. Immunol. 2008;180:5653e5661.

    Article  Google Scholar 

  21. Clayburgh DR, Barrett TA, Tang Y, Meddings JB, Van Eldik LJ, Watterson DM, Clarke LL, Mrsny RJ, Turner JR. Epithelial myosin light chain kinase-dependent barrier dysfunction mediates T cell activation- induced diarrhea in vivo. J. Clin. Invest. 2005;115:2702e2715.

    Article  Google Scholar 

  22. Ma TY, Boivin MA, Ye D, Pedram A, Said HM. Mechanism of TNF-{alpha} modulation of Caco-2 intestinal epithelial tight junction barrier: role of myosin light-chain kinase protein expression. Am J Physiol Gastrointest Liver Physiol. 2005;288(3):G422–30.

    Article  CAS  PubMed  Google Scholar 

  23. Cunningham KE, Turner JR. Myosin light chain kinase: pulling the strings of epithelial tight junction function. Ann N Y Acad Sci. 2012;1258:34–42.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  24. Ye D, Ma I, Ma TY. Molecular mechanism of tumor necrosis factor-alpha modulation of intestinal epithelial tight junction barrier. Am J Physiol Gastrointest Liver Physiol. 2006;290(3):G496–504.

    Article  CAS  PubMed  Google Scholar 

  25. Cao H, Zhou X, Zhang J, et al. Hydrogen sulfide protects against bleomycin-induced pulmonary fibrosis in rats by inhibiting NF-κB expression and regulating Th1/Th2 balance. Toxicol Lett. 2014;224(3):387–94.

    Article  CAS  PubMed  Google Scholar 

  26. Rao RK, Basuroy S, Rao VU, Karnaky KJ Jr, Gupta A. Tyrosine phosphorylation and dissociation of occludin-ZO-1 and E-cadherin-beta-catenin complexes from the cytoskeleton by oxidative stress. Biochem J. 2002;368(Pt 2):471–81.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Elias BC, Suzuki T, Seth A, Giorgianni F, Kale G, Shen L, et al. Phosphorylation of Tyr-398 and Tyr-402 in occludin prevents its interaction with ZO-1 and destabilizes its assembly at the tight junctions. J Biol Chem. 2009;284(3):1559–69.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  28. Murakami T, Felinski EA, Antonetti DA. Occludin phosphorylation and ubiquitination regulate tight junction trafficking and vascular endothelial growth factor-induced permeability. J Biol Chem. 2009;284(31):21036–46.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Fiorucci S, Distrutti E, Cirino G, Wallace JL. The emerging roles of hydrogen sulfide in the gastrointestinal tract and liver. Gastroenterology. 2006;131(1):259–71.

    Article  CAS  PubMed  Google Scholar 

  30. Zanardo RC, Brancaleone V, Distrutti E, et al. Hydrogen sulfide is an endogenous modulator of leukocyte-mediated inflammation. FASEB J. 2006;20:2118–20.

    Article  CAS  PubMed  Google Scholar 

  31. Wallace JL, Dicay M, McKnight W, et al. Hydrogen sulfide enhances ulcer healing in rats. FASEB J. 2007;21:4070–6.

    Article  CAS  PubMed  Google Scholar 

  32. Wallace JL, Blackler RW, Chan MV, et al. Anti-inflammatory and cytoprotective actions of hydrogen sulfide: translation to therapeutics. Antioxid Redox Signal. 2015;22:398–410.

    Article  CAS  PubMed  Google Scholar 

  33. Cao M, Wang P, Sun C, He W, Wang F. Amelioration of IFN-γ and TNF-α-induced intestinal epithelial barrier dysfunction by berberine via suppression of MLCK-MLC phosphorylation signaling pathway. PLoS ONE. 2013;8(5):e61944.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  34. Cheadle GA, Costantini TW, Lopez N, Bansal V, Eliceiri BP, Coimbra R. Enteric glia cells attenuate cytomix-induced intestinal epithelial barrier breakdown. PLoS ONE. 2013;8(7):e69042.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  35. Ye D, Ma TY. Cellular and molecular mechanisms that mediate basal and tumor necrosis factor-a induced regulation of myosin light chain kinase gene activity. J. Cell. Mol. Med. 2008;12:1331.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  36. Boivin MA, Ye D, Kennedy JC, Al-Sadi R, Shepela C, Ma TY. Mechanism of glucocorticoid regulation of the intestinal tight junction barrier. Am. J. Physiol. Gastrointest. Liver Physiol. 2007;292:590e598.

    Google Scholar 

  37. Moriez R, Salvador-Cartier C, Theodorou V, Fioramonti J, Eutamene H, Bueno L. Myosin light chain kinase is involved in lipopolysaccharide-induced disruption of colonic epithelial barrier and bacterial translocation in rats. Am. J. Pathol. 2005;167:1071e1079.

    Article  Google Scholar 

  38. Ma TY, Boivin MA, Ye D, Pedram A, Said HM. Mechanism of TNF-{alpha} modulation of Caco-2 intestinal epithelial tight junction barrier: role of myosin light-chain kinase protein expression. Am J Physiol Gastrointest Liver Physiol. 2005;288(3):G422–30.

    Article  CAS  PubMed  Google Scholar 

  39. Ma TY, Iwamoto GK, Hoa NT, Akotia V, Pedram A, Boivin MA, Said HM. TNF-alpha-induced increase in intestinal epithelial tight junction permeability requires NF-kappa B activation. Am J Physiol Gastrointest Liver Physiol. 2004;286(3):G367–76.

    Article  CAS  PubMed  Google Scholar 

  40. Guo C, Liang F, Shah Masood W, Yan X. Hydrogen sulfide protected gastric epithelial cell from ischemia/reperfusion injury by Keap1 s-sulfhydration, MAPK dependent anti-apoptosis and NF-κB dependent anti-inflammation pathway. Eur J Pharmacol. 2014;725:70–8.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

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

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Authors and Affiliations

  1. Division of General Surgery, Peking University First Hospital, Peking University, 8 Xi ShiKu Street, Beijing, 100034, People’s Republic of China

    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

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  1. Shan-wen Chen
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  2. Jing Zhu
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Correspondence to Peng-yuan Wang.

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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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Responsible Editor: John Di Battista.

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Chen, Sw., Zhu, J., Zuo, S. et al. Protective effect of hydrogen sulfide on TNF-α and IFN-γ-induced injury of intestinal epithelial barrier function in Caco-2 monolayers. Inflamm. Res. 64, 789–797 (2015). https://doi.org/10.1007/s00011-015-0862-5

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  • DOI: https://doi.org/10.1007/s00011-015-0862-5

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