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Fecal Supernatants from Active Ulcerative Colitis Patients Impair Colonic Epithelial Barrier Integrity and Activate Pelvic Mucosal Extrinsic Afferent Nerves
Author(s) -
Wardill Hannah R,
Dmochowska Nicole,
Campaniello Melissa A,
Mavrangelos Chris,
Bowen Joanne M,
Andrews Jane M,
Costello Sam P,
Hughes Patrick A
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.890.4
Subject(s) - ussing chamber , ulcerative colitis , feces , extracellular , colitis , dysbiosis , electrophysiology , chemistry , intestinal mucosa , tight junction , inflammation , gut flora , medicine , immunology , epithelium , pathology , biology , microbiology and biotechnology , biochemistry , disease
Ulcerative Colitis is characterised by colonic mucosal inflammation. Microbial dysbiosis is well documented in active UC, but little is understood regarding the physiological implications this has on epithelial barrier integrity or colonic extrinsic afferent signalling. We aimed to determine the effects of fecal supernatants from active UC on epithelial barrier and colo‐rectal mucosal extrinsic pelvic afferent nerves. Methods Excreted fecal samples from 10 patients with active UC (endoscopic MAYO subscore ≥2) were mixed with Ringers (Ussing) or Krebs solution (electrophysiology) at 1.5gm / 3 ml, centrifuged, and the supernatant filtered and pooled. Pooled fecal supernatants (FSN) were run over a 30kDa size exclusion column. Neat, high and low molecular weight FSN were applied to CACO‐2 monolayers or healthy mouse colon tissue in Ussing chamber experiments over a 2 hour period. Extracellular electrophysiological responses from pelvic colonic extrinsic afferent nerves were recorded from healthy mice. Mucosal afferents were characterised as responding to fine mucosal stroking with 10 mg Von Frey Hair. Neat FSN was applied to mucosal afferents for 5 min and mechanical sensitivity to graded mucosal stroking with Von Frey Hair re‐assessed. Results Neat UC FSN potently decreased the resistance of CACO‐2 monolayers and mouse colon over a 2 hour period compared to vehicle. Low molecular weight FSN also decreased the resistance of mouse colon, yet increased the resistance of CACO‐2 monolayers. High molecular weight FSN had no effect on either mouse tissue or CACO‐2 monolayers resistance. Neat UC FSN directly activated mucosal colonic afferent endings immediately following addition, but had no effect on mechanical sensitivity to mucosal stroking. Discussion Mediators present in faecal samples impair the resistance of the epithelial barrier and directly activate mucosal afferent nerve endings. The <30kDa fraction had the most potent activity on epithelial barrier. Further characterisation of the active mediator(s) and mechanisms has the potential to lead to new targets for the treatment of barrier dysfunction and nerve activation in active UC. Support or Funding Information Supported by NHMRC Australia