Reducing small intestinal permeability attenuates colitis in the IL10 gene-deficient mouse.
BACKGROUND
Defects in the small intestinal epithelial barrier have been associated with inflammatory bowel disease but their role in the causation of disease is still a matter of debate. In some models of disease increased permeability appears to be a very early event. The interleukin 10 (IL10) gene-deficient mouse spontaneously develops colitis after 12 weeks of age. These mice have been shown to have increased small intestinal permeability that appears early in life. Furthermore, the development of colitis is dependent upon luminal agents, as animals do not develop disease if raised under germ-free conditions.


OBJECTIVE
To determine if the elevated small bowel permeability can be prevented, and if by doing so colonic disease is prevented or attenuated.


METHODS
IL10 gene-deficient (IL10(-)/(-)) mice) were treated with AT-1001 (a zonulin peptide inhibitor), a small peptide previously demonstrated to reduce small intestinal permeability. Small intestinal permeability was measured, in vivo, weekly from 4 to 17 weeks of age. Colonic disease was assessed at 8 weeks in Ussing chambers, and at 17 weeks of age inflammatory cytokines and myeloperoxidase were measured in the colon. Colonic permeability and histology were also endpoints.


RESULTS
Treated animals showed a marked reduction in small intestinal permeability. Average area under the lactulose/mannitol time curve: 5.36 (SE 0.08) in controls vs 3.97 (SE 0.07) in the high-dose AT-1001 group, p<0.05. At 8 weeks of age there was a significant reduction of colonic mucosal permeability and increased electrical resistance. By 17 weeks of age, secretion of tumour necrosis factor alpha (TNFalpha) from a colonic explant was significantly lower in the treated group (25.33 (SE 4.30) pg/mg vs 106.93 (SE 17.51) pg/ml in controls, p<0.01). All other markers also demonstrated a clear reduction of colitis in the treated animals. Additional experiments were performed which demonstrated that AT-1001 was functionally active only in the small intestine.


CONCLUSIONS
This work suggests that increased intestinal permeability may be an important aetiological event in the development of colitis in IL10(-)/(-) mice.
Arrieta
M C
MC
Department of Medicine, University of Alberta, Walter C Mackenzie Health Science Ctre, Edmonton, AB, Canada.
Madsen
K
K
Doyle
J
J
Meddings
J
J
D000818:Animals; D002772:Cholera Toxin / Q000037:antagonists & inhibitors; D003092:Colitis / Q000276:immunology / Q000473:pathology / Q000503:physiopathology / Q000517:prevention & control*; D003106:Colon / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D015194:Diffusion Chambers, Culture; D006242:Haptoglobins; D016753:Interleukin-10 / Q000172:deficiency*; D007408:Intestinal Absorption / Q000187:drug effects*; D007421:Intestine, Small / Q000503:physiopathology*; D051379:Mice; D018345:Mice, Knockout; D009842:Oligopeptides / Q000627:therapeutic use; D010539:Permeability / Q000187:drug effects; D009195:Peroxidase / Q000378:metabolism; D011498:Protein Precursors
D016207:Cytokines; D006242:Haptoglobins; D009842:Oligopeptides; D011498:Protein Precursors; C408043:zonulin; D016753:Interleukin-10; D002772:Cholera Toxin; D009195:Peroxidase; C525167:larazotide acetate