Impaired intestinal barrier integrity in the colon of patients with irritable bowel syndrome: involvement of soluble mediators

Background: Growing evidence suggests that patients with irritable bowel syndrome (IBS) have increased intestinal permeability. In addition, mucosal soluble mediators are involved in the pathophysiology of pain in IBS. We aimed to investigate (1) paracellular permeability in colonic biopsies of patients with IBS; and (2) the ability of soluble factors from colonic biopsies to reproduce these alterations in vitro. Methods: Paracellular permeability in colonic biopsies of healthy subjects and patients with IBS was measured by mounting the biopsies in Ussing chambers. Cleared supernatant (SUP) of the culture from colonic biopsies was collected and applied to Caco-2 cells for 48 h. Paracellular permeability and transepithelial resistance (TER) were evaluated. mRNA expression of the tight junction proteins, zonula occludens (ZO)-1 and occludin, was assessed in colonic biopsies. Abdominal pain was assessed using a validated questionnaire. Results: Permeability of colonic biopsies was significantly higher in patients with IBS compared to healthy subjects. These changes were associated with significantly lower expression of ZO-1 mRNA in biopsies of IBS as compared to healthy subjects. Compared to healthy subjects, SUP of IBS markedly reduced TER and significantly increased permeability in Caco-2 cells. SUP of IBS patients induced a significant decrease of ZO-1 mRNA in Caco-2 as compared to healthy subjects. SUP-induced increased paracellular permeability correlated with the severity of abdominal pain. Conclusions: Our study shows that colonic soluble mediators are able to reproduce functional (permeability) and molecular (ZO-1 mRNA expression) alterations observed in IBS patients. These findings might pave the way both to identify novel biomarkers as well as new therapeutic targets in IBS.

[1]  T. Savidge,et al.  Neuro‐glial crosstalk in inflammatory bowel disease , 2008, Journal of internal medicine.

[2]  T. Wittmann,et al.  Increased faecal serine protease activity in diarrhoeic IBS patients: a colonic lumenal factor impairing colonic permeability and sensitivity , 2008, Gut.

[3]  S. P. Srinivas,et al.  Histamine-induced Myosin Light Chain Phosphorylation Breaks Down the Barrier Integrity of Cultured Corneal Epithelial Cells , 2007, Pharmaceutical Research.

[4]  M. Sofroniew,et al.  Enteric glia regulate intestinal barrier function and inflammation via release of S-nitrosoglutathione. , 2007, Gastroenterology.

[5]  M. Steinhoff,et al.  Role for protease activity in visceral pain in irritable bowel syndrome. , 2007, The Journal of clinical investigation.

[6]  R. Spiller,et al.  Abnormal Intestinal Permeability in Subgroups of Diarrhea-Predominant Irritable Bowel Syndromes , 2006, The American Journal of Gastroenterology.

[7]  G. Barbara.  Mucosal Barrier Defects in Irritable Bowel Syndrome. Who Left the Door Open? , 2006, The American Journal of Gastroenterology.

[8]  Hua Xie,et al.  Roles of histamine and its receptors in allergic and inflammatory bowel diseases. , 2005, World journal of gastroenterology.

[9]  J. Söderholm,et al.  Endoscopic biopsies in Ussing chambers evaluated for studies of macromolecular permeability in the human colon , 2005, Scandinavian journal of gastroenterology.

[10]  V. Théodorou,et al.  Acute stress-induced hypersensitivity to colonic distension depends upon increase in paracellular permeability: role of myosin light chain kinase , 2005, Pain.

[11]  W. Clark,et al.  Intestinal permeability in patients with irritable bowel syndrome after a waterborne outbreak of acute gastroenteritis in Walkerton, Ontario , 2004, Alimentary pharmacology & therapeutics.

[12]  D. Santini,et al.  Activated mast cells in proximity to colonic nerves correlate with abdominal pain in irritable bowel syndrome. , 2004, Gastroenterology.

[13]  J. Galmiche,et al.  Human ENS regulates the intestinal epithelial barrier permeability and a tight junction-associated protein ZO-1 via VIPergic pathways. , 2003, American journal of physiology. Gastrointestinal and liver physiology.

[14]  T. Munnik,et al.  Tumour necrosis factor α potentiates ion secretion induced by histamine in a human intestinal epithelial cell line and in mouse colon: involvement of the phospholipase D pathway , 2002, Gut.

[15]  P. Rampal,et al.  The putative role of inflammation in the irritable bowel syndrome , 2001, Gut.

[16]  S. Walsh,et al.  Neutrophil transmigration in inflammatory bowel disease is associated with differential expression of epithelial intercellular junction proteins. , 2001, The American journal of pathology.

[17]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[18]  S. Tsukita,et al.  Regulation of Tight Junction Permeability and Occludin Phosphorylation by RhoA-p160ROCK-dependent and -independent Mechanisms* , 2001, The Journal of Biological Chemistry.

[19]  R. Spiller,et al.  Increased rectal mucosal enteroendocrine cells, T lymphocytes, and increased gut permeability following acuteCampylobacter enteritis and in post-dysenteric irritable bowel syndrome , 2000, Gut.

[20]  J. Madara,et al.  Molecular physiology and pathophysiology of tight junctions. IV. Regulation of tight junctions by extracellular stimuli: nutrients, cytokines, and immune cells. , 2000, American journal of physiology. Gastrointestinal and liver physiology.

[21]  M. Itoh,et al.  Structural and signalling molecules come together at tight junctions. , 1999, Current opinion in cell biology.

[22]  K W Heaton,et al.  Functional bowel disorders and functional abdominal pain , 1999, Gut.

[23]  K. H. Peterson,et al.  Epithelial permeability to proteins in the noninflamed ileum of Crohn's disease? , 1999, Gastroenterology.

[24]  R. Naccarato,et al.  Discrepancies between reported food intolerance and sensitization test findings in irritable bowel syndrome patients , 1999, American Journal of Gastroenterology.

[25]  S. Crowe,et al.  Mast cell mediated ion transport in intestine from patients with and without inflammatory bowel disease , 1997, Gut.

[26]  E. Penner,et al.  Screening for celiac disease in first-degree relatives of patients with celiac disease by lactulose/mannitol test. , 1995, The American journal of gastroenterology.

[27]  N. Severs,et al.  No effect of histamine on human bronchial epithelial cell permeability and tight junctional integrity in vitro. , 1994, The European respiratory journal.

[28]  M. Zuckerman,et al.  Intestinal permeability to [51Cr]EDTA in infectious diarrhea , 1993, Digestive Diseases and Sciences.

[29]  C. Dupont,et al.  Modifications of Intestinal Permeability During Food Provocation Procedures in Pediatric Irritable Bowel Syndrome , 1990, Journal of pediatric gastroenterology and nutrition.

[30]  G. Sturniolo,et al.  [Irritable bowel syndrome]. , 1988, Giornale di clinica medica.

[31]  A. Axon,et al.  Intestinal permeability and screening tests for coeliac disease. , 1980, Gut.

[32]  N. Talley.  Increased faecal serine protease activity in diarrhoeic IBS patients: a colonic lumenal factor impairing colonic permeability and sensitivity , 2009 .

[33]  D. Katzka.  Mast Cell-Dependent Excitation of Visceral-Nociceptive Sensory Neurons in Irritable Bowel Syndrome , 2008 .

[34]  G. Barbara,et al.  Mast cell-dependent excitation of visceral-nociceptive sensory neurons in irritable bowel syndrome. , 2007, Gastroenterology.

[35]  D. Thakker,et al.  Modulation of the Tight Junctions of the Caco-2 Cell Monolayers by H2-antagonists , 2004, Pharmaceutical Research.

[36]  Thomas D. Schmittgen,et al.  Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2 2 DD C T Method , 2022 .