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Protease Activated Receptor-2 Induces Immune Activation and Visceral Hypersensitivity in Post-infectious Irritable Bowel Syndrome Mice

  • Lijun Du
  • Yanqin LongEmail author
  • John J. Kim
  • Binrui Chen
  • Yubin Zhu
  • Ning Dai
Original Article
  • 78 Downloads

Abstract

Background

The role of protease activated receptor-2 (PAR-2) in the pathogenesis of abdominal pain in irritable bowel syndrome (IBS) is not well defined.

Aims

To investigate the role of PAR-2-mediated visceral hypersensitivity in a post-infectious IBS (PI-IBS) mouse model.

Methods

T. spiralis-infected PI-IBS mouse model was used. Fecal serine protease activity and intestinal mast cells were evaluated. Intestinal permeability was assessed by urine lactulose/mannitol ratio, and colonic expressions of PAR-2 and tight junction (TJ) proteins were examined by Western blot. Intestinal immune profile was assessed by measuring Th (T helper) 1/Th2 cytokine expression. Visceral sensitivity was evaluated by abdominal withdrawal reflex in response to colorectal distention.

Results

Colonic PAR-2 expression as well as fecal serine protease activity and intestinal mast cell counts were elevated in PI-IBS compared to the control mice. Decreased colonic TJ proteins expression, increased lactulose/mannitol ratio, elevated colonic Th1/Th2 cytokine ratio, and visceral hypersensitivity were observed in PI-IBS compared to the control mice. Administration of PAR-2 agonist in control mice demonstrated similar changes observed in PI-IBS mice, while PAR-2 antagonist normalized the increased intestinal permeability and reduced visceral hypersensitivity observed in PI-IBS mice.

Conclusions

PAR-2 activation increases intestinal permeability leading to immune activation and visceral hypersensitivity in PI-IBS mouse model.

Keywords

Protease activated receptor-2 Post-infectious irritable bowel syndrome Intestinal permeability Tight junction T helper 

Notes

Acknowledgments

The study was granted and supported by the National Natural Science Foundation of China (No. 81200274, No. 81670487).

Author’s contribution

ND and YQL designed the research; LJD, YBZ, and BRC performed the research; LJD analyzed the data; LJD, YQL, and JJK wrote the paper; and YQL and JJK revised the paper. All authors approved the final manuscript.

Compliance with ethical standards

Conflict of interest

No competing interests to declare.

Supplementary material

10620_2018_5367_MOESM1_ESM.docx (1.2 mb)
Supplementary material 1 (DOCX 1226 kb)

References

  1. 1.
    Enck P, Aziz Q, Barbara G, et al. Irritable bowel syndrome. Nat Rev Dis Primers. 2016;2:16014.CrossRefGoogle Scholar
  2. 2.
    Spiller R, Garsed K. Postinfectious irritable bowel syndrome. Gastroenterology. 2009;136:1979–1988.CrossRefGoogle Scholar
  3. 3.
    Bueno L, Fioramonti J. Effects of inflammatory mediators on gut sensitivity. Can J Gastroenterol. 1999;13:42a–46a.CrossRefGoogle Scholar
  4. 4.
    Shulman RJ, Jarrett ME, Cain KC, et al. Associations among gut permeability, inflammatory markers, and symptoms in patients with irritable bowel syndrome. J Gastroenterol. 2014;49:1467–1476.CrossRefGoogle Scholar
  5. 5.
    Marshall JK, Thabane M, Garg AX, et al. Intestinal permeability in patients with irritable bowel syndrome after a waterborne outbreak of acute gastroenteritis in Walkerton, Ontario. Aliment Pharmacol Ther. 2004;20:1317–1322.CrossRefGoogle Scholar
  6. 6.
    Jacob C, Yang PC, Darmoul D, et al. Mast cell tryptase controls paracellular permeability of the intestine. Role of protease-activated receptor 2 and beta-arrestins. J Biol Chem. 2005;280:31936–31948.CrossRefGoogle Scholar
  7. 7.
    Bao Y, Hou W, Hua B. Protease-activated receptor 2 signalling pathways: a role in pain processing. Expert Opin Ther Targets. 2014;18:15–27.CrossRefGoogle Scholar
  8. 8.
    Barbara G, Wang B, Stanghellini V, et al. Mast cell-dependent excitation of visceral-nociceptive sensory neurons in irritable bowel syndrome. Gastroenterology. 2007;132:26–37.CrossRefGoogle Scholar
  9. 9.
    Buhner S, Li Q, Vignali S, et al. Activation of human enteric neurons by supernatants of colonic biopsy specimens from patients with irritable bowel syndrome. Gastroenterology. 2009;137:1425–1434.CrossRefGoogle Scholar
  10. 10.
    Lee JW, Park JH, Park DI, et al. Subjects with diarrhea-predominant IBS have increased rectal permeability responsive to tryptase. Dig Dis Sci. 2010;55:2922–2928.  https://doi.org/10.1007/s10620-009-1094-8.CrossRefPubMedGoogle Scholar
  11. 11.
    Rolland-Fourcade C, Denadai-Souza A, Cirillo C, et al. Epithelial expression and function of trypsin-3 in irritable bowel syndrome. Gut. 2017;66:1767–1778.CrossRefGoogle Scholar
  12. 12.
    Liang WJ, Zhang G, Luo HS, et al. Tryptase and protease-activated receptor 2 expression levels in irritable bowel syndrome. Gut Liver. 2015;10:382–390.PubMedCentralGoogle Scholar
  13. 13.
    Gecse K, Roka R, Ferrier L, et al. Increased faecal serine protease activity in diarrhoeic IBS patients: a colonic lumenal factor impairing colonic permeability and sensitivity. Gut. 2008;57:591–599.CrossRefGoogle Scholar
  14. 14.
    Institute for Laboratory Animal Research. Guide for the Care and Use of Laboratory Animals. 8th ed. Washington, DC: National Academies Press; 2011.Google Scholar
  15. 15.
    National Research Council Subcommittee on Laboratory Animal. Nutrient Requirements of Laboratory Animals. 4th ed Washington, DC: National Academies Press; 1995.Google Scholar
  16. 16.
    Bercik P, Wang L, Verdu EF, et al. Visceral hyperalgesia and intestinal dysmotility in a mouse model of postinfective gut dysfunction. Gastroenterology. 2004;127:179–187.CrossRefGoogle Scholar
  17. 17.
    Cenac N, Garcia-Villar R, Ferrier L, et al. Proteinase-activated receptor-2-induced colonic inflammation in mice: possible involvement of afferent neurons, nitric oxide, and paracellular permeability. J Immunol. 2003;170:4296–4300.CrossRefGoogle Scholar
  18. 18.
    Nguyen C, Coelho AM, Grady E, et al. Colitis induced by proteinase-activated receptor-2 agonists is mediated by a neurogenic mechanism. Can J Physiol Pharmacol. 2003;81:920–927.CrossRefGoogle Scholar
  19. 19.
    Han SK, Dong X, Hwang JI, et al. Orphan G protein-coupled receptors MrgA1 and MrgC11 are distinctively activated by RF-amide-related peptides through the Galpha q/11 pathway. Proc Natl Acad Sci USA. 2002;99:14740–14745.CrossRefGoogle Scholar
  20. 20.
    Park Y, Yang J, Zhang H, et al. Effect of PAR2 in regulating TNF-alpha and NAD(P)H oxidase in coronary arterioles in type 2 diabetic mice. Basic Res Cardiol. 2011;106:111–123.CrossRefGoogle Scholar
  21. 21.
    Al-Ani B, Saifeddine M, Wijesuriya SJ, et al. Modified proteinase-activated receptor-1 and -2 derived peptides inhibit proteinase-activated receptor-2 activation by trypsin. J Pharmacol Exp Ther. 2002;300:702–708.CrossRefGoogle Scholar
  22. 22.
    Kamp EH, Jones RC 3rd, Tillman SR, et al. Quantitative assessment and characterization of visceral nociception and hyperalgesia in mice. Am J Physiol Gastrointest Liver Physiol. 2003;284:G434–G444.CrossRefGoogle Scholar
  23. 23.
    Al-Chaer ED, Kawasaki M, Pasricha PJ. A new model of chronic visceral hypersensitivity in adult rats induced by colon irritation during postnatal development. Gastroenterology. 2000;119:1276–1285.CrossRefGoogle Scholar
  24. 24.
    Lan C, Sun XN, Zhou XC, et al. Preinduced intestinal HSP70 improves visceral hypersensitivity and abnormal intestinal motility in PI-IBS mouse model. Asian Pac J Trop Med. 2016;9:302–305.CrossRefGoogle Scholar
  25. 25.
    Long Y, Liu Y, Tong J, et al. Effectiveness of trimebutine maleate on modulating intestinal hypercontractility in a mouse model of postinfectious irritable bowel syndrome. Eur J Pharmacol. 2010;636:159–165.CrossRefGoogle Scholar
  26. 26.
    Krawisz JE, Sharon P, Stenson WF. Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Assessment of inflammation in rat and hamster models. Gastroenterology. 1984;87:1344–1350.PubMedGoogle Scholar
  27. 27.
    Barbara G, Vallance BA, Collins SM. Persistent intestinal neuromuscular dysfunction after acute nematode infection in mice. Gastroenterology. 1997;113:1224–1232.CrossRefGoogle Scholar
  28. 28.
    Gecse K, Roka R, Ferrier L, et al. Increased faecal serine protease activity in diarrhoeic IBS patients: a colonic lumenal factor impairing colonic permeability and sensitivity. Gut. 2008;57:591–599.CrossRefGoogle Scholar
  29. 29.
    Bertiaux-Vandaële N, Youmba SB, Belmonte L, et al. The expression and the cellular distribution of the tight junction proteins are altered in irritable bowel syndrome patients with differences according to the disease subtype. Am J Gastroenterol. 2011;106:2165–2173.CrossRefGoogle Scholar
  30. 30.
    Kong W, McConalogue K, Khitin LM, et al. Luminal trypsin may regulate enterocytes through proteinase-activated receptor 2. Proc Natl Acad Sci USA. 1997;94:8884–8889.CrossRefGoogle Scholar
  31. 31.
    Wang P, Chen FX, Du C, et al. Increased production of BDNF in colonic epithelial cells induced by fecal supernatants from diarrheic IBS patients. Sci Rep. 2015;5:10121.CrossRefGoogle Scholar
  32. 32.
    Coelho AM, Vergnolle N, Guiard B, et al. Proteinases and proteinase-activated receptor 2: a possible role to promote visceral hyperalgesia in rats. Gastroenterology. 2002;122:1035–1047.CrossRefGoogle Scholar
  33. 33.
    Kawao N, Ikeda H, Kitano T, et al. Modulation of capsaicin-evoked visceral pain and referred hyperalgesia by protease-activated receptors 1 and 2. J Pharmacol Sci. 2004;94:277–285.CrossRefGoogle Scholar
  34. 34.
    Valdez-Morales EE, Overington J, Guerrero-Alba R, et al. Sensitization of peripheral sensory nerves by mediators from colonic biopsies of diarrhea-predominant irritable bowel syndrome patients: a role for PAR2. Am J Gastroenterol. 2013;108:1634–1643.CrossRefGoogle Scholar
  35. 35.
    Jinguji Y, Ishikawa H. Electron microscopic observations on the maintenance of the tight junction during cell division in the epithelium of the mouse small intestine. Cell Struct Funct. 1992;17:27–37.CrossRefGoogle Scholar
  36. 36.
    Al-Sadi R, Boivin M, Ma T. Mechanism of cytokine modulation of epithelial tight junction barrier. Front Biosci (Landmark Ed). 2009;14:2765–2778.CrossRefGoogle Scholar
  37. 37.
    Al-Sadi R, Ye D, Dokladny K, et al. Mechanism of IL-1beta-induced increase in intestinal epithelial tight junction permeability. J Immunol. 2008;180:5653–5661.CrossRefGoogle Scholar
  38. 38.
    He F, Peng J, Deng XL, et al. Mechanisms of tumor necrosis factor-alpha-induced leaks in intestine epithelial barrier. Cytokine. 2012;59:264–272.CrossRefGoogle Scholar
  39. 39.
    Bertiaux-Vandaele N, Youmba SB, Belmonte L, et al. The expression and the cellular distribution of the tight junction proteins are altered in irritable bowel syndrome patients with differences according to the disease subtype. Am J Gastroenterol. 2011;106:2165–2173.CrossRefGoogle Scholar
  40. 40.
    Liebregts T, Adam B, Bredack C, et al. Immune activation in patients with irritable bowel syndrome. Gastroenterology. 2007;132:913–920.CrossRefGoogle Scholar
  41. 41.
    Ohman L, Isaksson S, Lindmark AC, et al. T-cell activation in patients with irritable bowel syndrome. Am J Gastroenterol. 2009;104:1205–1212.CrossRefGoogle Scholar
  42. 42.
    Lee KJ, Kim YB, Kim JH, et al. The alteration of enterochromaffin cell, mast cell, and lamina propria T lymphocyte numbers in irritable bowel syndrome and its relationship with psychological factors. J Gastroenterol Hepatol. 2008;23:1689–1694.CrossRefGoogle Scholar
  43. 43.
    Chen J, Zhang Y, Deng Z. Imbalanced shift of cytokine expression between T helper 1 and T helper 2 (Th1/Th2) in intestinal mucosa of patients with post-infectious irritable bowel syndrome. BMC Gastroenterol. 2012;12:91.CrossRefGoogle Scholar
  44. 44.
    Hyun E, Andrade-Gordon P, Steinhoff M, et al. Protease-activated receptor-2 activation: a major actor in intestinal inflammation. Gut. 2008;57:1222–1229.CrossRefGoogle Scholar
  45. 45.
    Cenac N, Coelho AM, Nguyen C, et al. Induction of intestinal inflammation in mouse by activation of proteinase-activated receptor-2. Am J Pathol. 2002;161:1903–1915.CrossRefGoogle Scholar
  46. 46.
    Wang F, Graham WV, Wang Y, et al. Interferon-gamma and tumor necrosis factor-alpha synergize to induce intestinal epithelial barrier dysfunction by up-regulating myosin light chain kinase expression. Am J Pathol. 2005;166:409–419.CrossRefGoogle Scholar
  47. 47.
    Richardson JD, Vasko MR. Cellular mechanisms of neurogenic inflammation. J Pharmacol Exp Ther. 2002;302:839–845.CrossRefGoogle Scholar
  48. 48.
    Park JH, Rhee PL, Kim HS, et al. Mucosal mast cell counts correlate with visceral hypersensitivity in patients with diarrhea predominant irritable bowel syndrome. J Gastroenterol Hepatol. 2006;21:71–78.CrossRefGoogle Scholar
  49. 49.
    Song J, Zhang L, Bai T, et al. Mast cell-dependent mesenteric afferent activation by mucosal supernatant from different bowel segments of guinea pigs with post-infectious irritable bowel syndrome. J Neurogastroenterol Motil. 2015;21:236–246.CrossRefGoogle Scholar
  50. 50.
    Dothel G, Barbaro MR, Boudin H, et al. Nerve fiber outgrowth is increased in the intestinal mucosa of patients with irritable bowel syndrome. Gastroenterology. 2015;148:1002–1011.e1004.CrossRefGoogle Scholar
  51. 51.
    Long Y, Du L, Kim JJ, et al. MLCK-mediated intestinal permeability promotes immune activation and visceral hypersensitivity in PI-IBS mice. Neurogastroenterol Motil. 2018;30:e13348.CrossRefGoogle Scholar
  52. 52.
    Liu Q, Weng HJ, Patel KN, et al. The distinct roles of two GPCRs, MrgprC11 and PAR2, in itch and hyperalgesia. Sci Signal. 2011;4:ra45.PubMedPubMedCentralGoogle Scholar
  53. 53.
    Hockley JRF, Taylor TS, Callejo G, et al. Single-cell RNAseq reveals seven classes of colonic sensory neuron. Gut. 2018;0:1–12.Google Scholar
  54. 54.
    Langford DJ, Bailey AL, Chanda ML, et al. Coding of facial expressions of pain in the laboratory mouse. Nat Methods. 2010;7:447–449.CrossRefGoogle Scholar
  55. 55.
    Christianson JA, Gebhart GF. Assessment of colon sensitivity by luminal distension in mice. Nat Protoc. 2007;2:2624–2631.CrossRefGoogle Scholar
  56. 56.
    Klem F, Wadhwa A, Prokop LJ, et al. Prevalence, risk factors, and outcomes of irritable bowel syndrome after infectious enteritis: a systematic review and meta-analysis. Gastroenterology. 2017;152:1042–1054.e1041.CrossRefGoogle Scholar
  57. 57.
    Heitkemper MM, Chang L. Do fluctuations in ovarian hormones affect gastrointestinal symptoms in women with irritable bowel syndrome? Gend Med. 2009;6:152–167.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Gastroenterology, School of Medicine, Sir Run Run Shaw HospitalZhejiang UniversityHangzhouChina
  2. 2.Division of Gastroenterology & HepatologyLoma Linda University HealthLoma LindaUSA

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