Journal Article

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

Portier, Guillaume, Deraison, Celine, Denadai-Souza, Alexandre, Jaramillo, Josue Obed, Bonnet, Delphine, Vergnolle, Nathalie, Motta, Jean-Paul, Neunlist, Michel, Vanner, Stephen, Desormeaux, Cleo, Berghe, Pieter Vanden, Taché, Yvette, Coron, Emmanuel, Larauche, Muriel, Alric, Laurent, Lopez, Cintya, Cenac, Nicolas, Rolland-Fourcade, Claire, Cirillo, Carla, Kirzin, Sylvain

2017

Overview

ObjectivesProteases 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.DesignTrypsin-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 Ca2+ 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.ResultsWe 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.ConclusionsIn 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.

• Publisher: BMJ Publishing Group,CCSD,BMJ Publishing Group LTD
• Subject: Animals / Caco-2 Cells / Calcium imaging / Case-Control Studies / Colon / Colon - enzymology / Colon - innervation / Culture Media, Conditioned - pharmacology / Dipeptides - pharmacology / Distension / Enteric nervous system / Enteric Nervous System - cytology / Enteric Nervous System - diagnostic imaging / Enteric Nervous System - drug effects / Epithelial cells / Epithelial Cells - drug effects / Epithelial Cells - enzymology / Epithelium / Female / Ganglia, Spinal - cytology / Human health and pathology / Humans / Hypersensitivity / Hypersensitivity - enzymology / Hépatology and Gastroenterology / Immunohistochemistry / Intestinal Mucosa - drug effects / Intestinal Mucosa - enzymology / Intestinal Mucosa - pathology / Intestine / Irritable bowel syndrome / Irritable Bowel Syndrome - enzymology / Irritable Bowel Syndrome - genetics / Isoforms / Isoxazoles - pharmacology / Life Sciences / Lipopolysaccharides - pharmacology / Male / Mice / Microscopy, Confocal / Nerves / Neurogastroenterology / Neurons, Afferent - drug effects / Neurons, Afferent - physiology / Pain / Par-2 protein / Permeability - drug effects / Proteases / Rats / Receptor, PAR-2 - antagonists & inhibitors / Receptor, PAR-2 - metabolism / Reverse transcription / RNA, Messenger - analysis / Rodents / Sensory neurons / Small intestine / Trypsin / Trypsin - genetics / Trypsin - metabolism / Trypsin - pharmacology / Trypsinogen - genetics / Up-Regulation
• ISBN: 0004093638000