Stress-induced disruption of colonic epithelial barrier: role of interferon-gamma and myosin light chain kinase in mice.
OBJECTIVE
Stressful life events are supposed to be involved in various diseases such as inflammatory bowel diseases and irritable bowel syndrome. Impairment of the intestinal epithelial barrier function is a suspected consequence of stress, but the underlying mechanisms remain unclear. This study aimed to determine the mechanisms through which stress modulates the colonic epithelial barrier.


METHODS
Cytokine messenger RNA (mRNA) expression was evaluated in murine colon, liver, and spleen by competitive reverse-transcription polymerase chain reaction after 1-4 days of daily 2-hour stress sessions. Colonic paracellular permeability was measured as the in vivo lumen-to-blood ratio of (51)Cr-ethylenediaminetetraacetic acid. The effect of a myosin light chain (MLC) kinase inhibitor (ML-7) was assessed on stress-induced interferon (IFN)-gamma mRNA expression and colonic epithelial barrier impairment, and MLC phosphorylation was determined by immunoblot. Finally, the incidence of repeated stress sessions on bacterial translocation was determined.


RESULTS
Repeated stress induced an overexpression of colonic IFN-gamma. In the liver, higher levels of IFN-gamma, interleukin (IL)-4, and IL-10 mRNAs were detected and were associated with bacterial translocation, inflammation, and apoptosis. Stress increased colonic permeability of control mice, but not of SCID and IFN-gamma-deficient mice. ML-7 inhibited the stress-induced increased permeability, bacterial translocation, and cytokine overexpression in the liver and restored a normal histology. Larger amounts of phosphorylated MLC were detected in stressed animals.


CONCLUSIONS
Repeated stress sessions drive organ-specific cytokine expression patterns and alter colonic mucosal barrier functions associated with bacterial translocation. This effect depends on the presence of CD4(+) T cells and requires IFN-gamma production and MLC phosphorylation.
Ferrier
Laurent
L
Neuro-Gastroenterology and Nutrition Unit, Institut National de la Recherche Agronomique, Toulouse, France.
Mazelin
Ludmilla
L
Cenac
Nicolas
N
Desreumaux
Pierre
P
Janin
Anne
A
Emilie
Dominique
D
Colombel
Jean-Frederic
JF
Garcia-Villar
Rafael
R
Fioramonti
Jean
J
Bueno
Lionel
L
D016475:3T3 Cells; D000818:Animals; D003106:Colon / Q000378:metabolism*; D016207:Cytokines / Q000235:genetics; D005260:Female; D007371:Interferon-gamma / Q000502:physiology*; D007413:Intestinal Mucosa / Q000378:metabolism; D008099:Liver / Q000473:pathology; D051379:Mice; D016513:Mice, SCID; D009219:Myosin-Light-Chain Kinase / Q000502:physiology*; D009928:Organ Specificity; D010539:Permeability; D013154:Spleen / Q000473:pathology; D013312:Stress, Physiological / Q000378:metabolism*
D016207:Cytokines; D007371:Interferon-gamma; D009219:Myosin-Light-Chain Kinase