Serine proteases as luminal mediators of intestinal barrier dysfunction and symptom severity in IBS.
Adult
Animals
Biopsy
Caco-2 Cells
Case-Control Studies
Colon
/ pathology
Dysbiosis
/ enzymology
Feces
/ enzymology
Female
Gastrointestinal Microbiome
Humans
Intestinal Absorption
/ physiology
Intestinal Mucosa
/ metabolism
Irritable Bowel Syndrome
/ enzymology
Male
Mice
Middle Aged
Permeability
Prospective Studies
Proteolysis
Serine Proteases
/ physiology
Severity of Illness Index
Tight Junction Proteins
/ metabolism
Campylobacter
gastroenteritis
germ-free mice
microbiome
trypsin
Journal
Gut
ISSN: 1468-3288
Titre abrégé: Gut
Pays: England
ID NLM: 2985108R
Informations de publication
Date de publication:
01 2020
01 2020
Historique:
received:
16
08
2018
revised:
13
03
2019
accepted:
16
03
2019
pubmed:
30
3
2019
medline:
18
12
2019
entrez:
30
3
2019
Statut:
ppublish
Résumé
The intestinal lumen contains several proteases. Our aim was to determine the role of faecal proteases in mediating barrier dysfunction and symptoms in IBS. 39 patients with IBS and 25 healthy volunteers completed questionnaires, assessments of in vivo permeability, ex vivo colonic barrier function in Ussing chambers, tight junction (TJ) proteins, ultrastructural morphology and 16 s sequencing of faecal microbiota rRNA. A casein-based assay was used to measure proteolytic activity (PA) in faecal supernatants (FSNs). Colonic barrier function was determined in mice (ex-germ free) humanised with microbial communities associated with different human PA states. Patients with IBS had higher faecal PA than healthy volunteers. 8/20 postinfection IBS (PI-IBS) and 3/19 constipation- predominant IBS had high PA (>95th percentile). High-PA patients had more and looser bowel movements, greater symptom severity and higher in vivo and ex vivo colonic permeability. High-PA FSNs increased paracellular permeability, decreased occludin and increased phosphorylated myosin light chain (pMLC) expression. Serine but not cysteine protease inhibitor significantly blocked high-PA FSN effects on barrier. The effects on barrier were diminished by pharmacological or siRNA inhibition of protease activated receptor-2 (PAR-2). Patients with high-PA IBS had lower occludin expression, wider TJs on biopsies and reduced microbial diversity than patients with low PA. Mice humanised with high-PA IBS microbiota had greater in vivo permeability than those with low-PA microbiota. A subset of patients with IBS, especially in PI-IBS, has substantially high faecal PA, greater symptoms, impaired barrier and reduced microbial diversity. Commensal microbiota affects luminal PA that can influence host barrier function.
Identifiants
pubmed: 30923071
pii: gutjnl-2018-317416
doi: 10.1136/gutjnl-2018-317416
pmc: PMC6765451
mid: NIHMS1529240
doi:
Substances chimiques
Tight Junction Proteins
0
Serine Proteases
EC 3.4.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
62-73Subventions
Organisme : NIDDK NIH HHS
ID : K23 DK103911
Pays : United States
Organisme : NIDDK NIH HHS
ID : P30 DK084567
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK114007
Pays : United States
Organisme : NIDDK NIH HHS
ID : R03 DK120745
Pays : United States
Informations de copyright
© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.
Déclaration de conflit d'intérêts
Competing interests: MG has served on the advisory board or received research support from Takeda, DongA, Ironwood and Napo.
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