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Recent studies showed that patients with diarrhea-predominant irritable bowel syndrome (IBS-D) had an increased intestinal permeability as well as a decreased expression of tight junctions. Glutamine, the major substrate of rapidly dividing cells, is able to modulate intestinal permeability and tight junction expression in other diseases. We aimed to evaluate, ex vivo, glutamine effects on tight junction proteins, claudin-1 and occludin, in the colonic mucosa of patients with IBS-D.Twelve patients with IBS-D, diagnosed with the Rome III criteria, were included (8 women/4 men, aged 40.7 ± 6.9 years). Colonic biopsy specimens were collected and immediately incubated for 18 hours in culture media with increasing concentrations of glutamine from 0.6-10 mmol/L. Claudin-1 and occludin expression was then measured by immunoblot, and concentrations of cytokines were assessed by multiplex technology. Claudin-1 expression was affected by glutamine (P.05, analysis of variance). In particularly, 10 mmol/L glutamine increased claudin-1 expression compared with 0.6 mmol/L glutamine (0.47 ± 0.04 vs 0.33 ± 0.03, P.05). In contrast, occludin expression was not significantly modified by glutamine. Interestingly, glutamine effect was negatively correlated to claudin-1 (Pearson r = -0.83, P.001) or occludin basal expression (Pearson r = -0.84, P.001), suggesting that glutamine had more marked effects when tight junction protein expression was altered. Cytokine concentrations in culture media were not modified by glutamine treatment.Glutamine increased claudin-1 expression in the colonic mucosa of patients with IBS-D. In addition, glutamine effect seems to be dependent on basal expression of tight junction proteins.