Background & aims: Antibiotic (ABx) therapy is associated with increased risk for Crohn's disease but underlying mechanisms are unknown. We observed high fecal serine protease activity (PA) to be a frequent side effect of ABx therapy. The aim of the present study was to unravel whether this rise in large intestinal PA may promote colitis development via detrimental effects on the large intestinal barrier.
Methods: Transwell experiments were used to assess the impact of high PA in ABx-treated patients or vancomycin/metronidazole-treated mice on the epithelial barrier. Serine protease profiling was performed using liquid chromatography-mass spectrometry/mass spectrometry analysis. The impact of high large intestinal PA on the intestinal barrier in wild-type and interleukin (IL)10-/- mice and on colitis development in IL10-/- mice was investigated using vancomycin/metronidazole with or without oral serine protease inhibitor (AEBSF) treatment.
Results: The ABx-induced, high large intestinal PA was caused by significantly increased levels of pancreatic proteases and impaired epithelial barrier integrity. In wild-type mice, the rise in PA caused a transient increase in intestinal permeability but did not affect susceptibility to chemically induced acute colitis. In IL10-/- mice, increased PA caused a consistent impairment of the intestinal barrier associated with inflammatory activation in the large intestinal tissue. In the long term, the vancomycin/metronidazole-induced lasting increase in PA aggravated colitis development in IL10-/- mice.
Conclusions: High large intestinal PA is a frequent adverse effect of ABx therapy, which is detrimental to the large intestinal barrier and may contribute to the development of chronic intestinal inflammation in susceptible individuals.
Keywords: ABx, antibiotics; AEBSF, 4-(2-aminoethyl) benzenesulfonyl fluoride hydrochloride; DSS, dextran sulfate sodium; Epithelial Barrier; GF, germ-free; Gut Microbiota; IBD, inflammatory bowel diseases; IL, interleukin; Inflammatory Bowel Diseases; LC-MS/MS, liquid chromatography–mass spectrometry/mass spectrometry; PA, protease activity; PBS, phosphate-buffered saline; PMSF, phenylmethane-sulfonylfluoride; SPF, specific pathogen-free; Serine Proteases; TEER, transepithelial electrical resistance; V/M, vancomycin/metronidazole; WT, wild-type; cecal-sup, cecal-supernatants; ctr, control; stool-sup, stool-supernatants.