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Epithelial expression and function of trypsin-3 in irritable bowel syndrome

Published on Oct 1, 2017in Gut17.943
· DOI :10.1136/gutjnl-2016-312094
Claire Rolland-Fourcade1
Estimated H-index: 1
(University of Toulouse),
Alexandre Denadai-Souza7
Estimated H-index: 7
(University of Toulouse)
+ 17 AuthorsNathalie Vergnolle62
Estimated H-index: 62
(University of Toulouse)
Sources
Abstract
Objectives Proteases are key mediators of pain and altered enteric neuronal signalling, although the types and sources of these important intestinal mediators are unknown. We hypothesised that intestinal epithelium is a major source of trypsin-like activity in patients with IBS and this activity signals to primary afferent and enteric nerves and induces visceral hypersensitivity. Design Trypsin-like activity was determined in tissues from patients with IBS and in supernatants of Caco-2 cells stimulated or not. These supernatants were also applied to cultures of primary afferents. mRNA isoforms of trypsin ( PRSS1 , 2 and 3 ) were detected by reverse transcription-PCR, and trypsin-3 protein expression was studied by western blot analysis and immunohistochemistry. Electrophysiological recordings and Ca 2+ imaging in response to trypsin-3 were performed in mouse primary afferent and in human submucosal neurons, respectively. Visceromotor response to colorectal distension was recorded in mice administered intracolonically with trypsin-3. Results We showed that stimulated intestinal epithelial cells released trypsin-like activity specifically from the basolateral side. This activity was able to activate sensory neurons. In colons of patients with IBS, increased trypsin-like activity was associated with the epithelium. We identified that trypsin-3 was the only form of trypsin upregulated in stimulated intestinal epithelial cells and in tissues from patients with IBS. Trypsin-3 was able to signal to human submucosal enteric neurons and mouse sensory neurons, and to induce visceral hypersensitivity in vivo, all by a protease-activated receptor-2-dependent mechanism. Conclusions In IBS, the intestinal epithelium produces and releases the active protease trypsin-3, which is able to signal to enteric neurons and to induce visceral hypersensitivity.
  • References (26)
  • Citations (26)
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References26
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#1Ami D. Sperber (BGU: Ben-Gurion University of the Negev)H-Index: 27
#2Dan L. DumitrascuH-Index: 12
Last. William E. Whitehead (UNC: University of North Carolina at Chapel Hill)H-Index: 75
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Objectives The global prevalence of IBS is difficult to ascertain, particularly in light of the heterogeneity of published epidemiological studies. The aim was to conduct a literature review, by experts from around the world, of community-based studies on IBS prevalence. Design Searches were conducted using predetermined search terms and eligibility criteria, including papers in all languages. Pooled prevalence rates were calculated by combining separate population survey prevalence estimates to...
79 CitationsSource
#1Nabila Moussaoui (UCLA: University of California, Los Angeles)H-Index: 3
#2Muriel H. Larauche (UCLA: University of California, Los Angeles)H-Index: 20
Last. Yvette Taché (UCLA: University of California, Los Angeles)H-Index: 71
view all 7 authors...
A few studies indicate that limited nesting stress (LNS) alters maternal behavior and the hypothalamic pituitary adrenal (HPA) axis of dams and offspring in male Sprague Dawley rats. In the present study, we evaluated the impact of LNS on maternal behavior in Wistar rats, and on the HPA axis, glycemia and in vivo intestinal permeability of male and female offspring. Intestinal permeability is known to be elevated during the first week postnatally and influenced by glucocorticoids. Dams and neona...
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#1Alexandre P. Alloy (Mayo Clinic)H-Index: 2
#2Olumide Kayode (Mayo Clinic)H-Index: 4
Last. Evette S. Radisky (Mayo Clinic)H-Index: 25
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Abstract Human mesotrypsin is highly homologous to other mammalian trypsins, and yet is functionally unique in possessing resistance to inhibition by canonical serine protease inhibitors and in cleaving these inhibitors as preferred substrates. Arg-193 and Ser-39 have been identified as contributors to the inhibitor resistance and cleavage capability of mesotrypsin, but it is not known whether these residues fully account for the unusual properties of mesotrypsin. Here we use human cationic tryp...
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#1Nicolas Cenac (Paul Sabatier University)H-Index: 29
#2Tereza Bautzova (Paul Sabatier University)H-Index: 5
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Background & Aims In mice, activation of the transient receptor potential cation channels (TRP) TRPV1, TRPV4, and TRPA1 causes visceral hypersensitivity. These receptors and their agonists might be involved in development of irritable bowel syndrome (IBS). We investigated whether polyunsaturated fatty acid (PUFA) metabolites, which activate TRPs, are present in colon tissues from patients with IBS and act as endogenous agonists to induce hypersensitivity. Methods We analyzed colon biopsy samples...
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#1David GrundmannH-Index: 5
#2Markus KlotzH-Index: 11
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The enteric nervous system (ENS) orchestrates a broad range of important gastrointestinal functions such as intestinal motility and gastric secretion. The ENS can be affected by environmental factors, diet and disease. Changes due to these alterations are often hard to evaluate in detail when whole gut samples are used. Analyses based on pure ENS tissue can more effectively reflect the ongoing changes during pathological processes. Here, we present an optimized approach for the isolation of pure...
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Mesotrypsin is an isoform of trypsin that is uniquely resistant to polypeptide trypsin inhibitors and can cleave some inhibitors rapidly. Previous studies have shown that the amyloid precursor protein Kunitz protease inhibitor domain (APPI) is a specific substrate of mesotrypsin and that stabilization of the APPI cleavage site in a canonical conformation contributes to recognition by mesotrypsin. We hypothesized that other proteins possessing potential cleavage sites stabilized in a similar conf...
17 CitationsSource
#1David Tooth (University of Nottingham)H-Index: 12
#2K Garsed (University of Nottingham)H-Index: 13
Last. Robin C. Spiller (University of Nottingham)H-Index: 69
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Objectives Faecal serine proteases (FSPs) may play a role in irritable bowel syndrome with diarrhoea (IBS-D), but their origin is unclear. We aimed to structurally characterise them and define the impact of colonic cleansing and transit time. Design Faecal samples were obtained from 30 healthy volunteers (HV) and 79 patients with IBS-D participating in a trial of ondansetron versus placebo. Colonic transit was measured using radio-opaque markers. Samples were also obtained from 24 HV before and ...
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Sensitization of Peripheral Sensory Nerves by Mediators From Colonic Biopsies of Diarrhea-Predominant Irritable Bowel Syndrome Patients: A Role for PAR2
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#1Muriel H. Larauche (UCLA: University of California, Los Angeles)H-Index: 20
#2Agata Mulak (UCLA: University of California, Los Angeles)H-Index: 9
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Background Chronic psychological stress-induced alterations in visceral sensitivity have been predominantly assessed in male rodents. We investigated the effect of acute and repeated water avoidance stress (WAS) on the visceromotor response (VMR) to colorectal distension (CRD) and the role of opioids in male and cycling female Wistar rats using a novel non-invasive manometric technique. Methods After a baseline VMR (1st CRD, day 0), rats were exposed to WAS (1 h day−1) either once or for four co...
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Elafin, a natural protease inhibitor expressed in healthy intestinal mucosa, has pleiotropic anti-inflammatory properties in vitro and in animal models. We found that mucosal expression of Elafin is diminished in patients with inflammatory bowel disease (IBD). This defect is associated with increased elastolytic activity (elastase-like proteolysis) in colon tissue. Weengineeredtwofood-grade strainsof lacticacidbacteria (LAB)to express anddeliverElafin tothesite ofinflammationinthecolontoassessth...
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