Original Article
Copyright ©2014 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 14, 2014; 20(22): 6832-6843
Published online Jun 14, 2014. doi: 10.3748/wjg.v20.i22.6832
Multispecies probiotic protects gut barrier function in experimental models
Mylene Nébot-Vivinus, Cherryl Harkat, Hanene Bzioueche, Christel Cartier, Raffaella Plichon-Dainese, Lara Moussa, Helene Eutamene, Dorsa Pishvaie, Sophie Holowacz, Christian Seyrig, Thierry Piche, Vassilia Theodorou
Mylene Nébot-Vivinus, Hanene Bzioueche, Department of Immunology, Pole of Biology, CHU de Nice, 06103 Nice, France
Mylene Nébot-Vivinus, Hanene Bzioueche, Raffaella Plichon-Dainese, Thierry Piche, EA 6302 “Tolérance immunitaire”, Université de Nice Sophia Antipolis, 06103 Nice, France
Cherryl Harkat, Christel Cartier, Lara Moussa, Helene Eutamene, Vassilia Theodorou, Neuro-Gastroenterology and Nutrition Group, Toxalim UMR 1331 INRA/INPT/UPS, F-31027 Toulouse, France
Raffaella Plichon-Dainese, Dorsa Pishvaie, Thierry Piche, Department of Gastroenterology, CHU de Nice, 06103 Nice, France
Sophie Holowacz, Christian Seyrig, Groupe PiLeJe, 37 Quai de Grenelle, 75015 Paris Cedex 15, France
Author contributions: Piche T and Theodorou V participated equally in the work; Nébot-Vivinus M, Piche T and Theodorou V designed the research (project conception, development of the overall research plan and supervision of the study) and wrote the paper; Harkat C, Bzioueche H, Cartier C, Moussa L, Pichon-Dainese R, Eutamene H conducted the research (performed the experiments and collected data); Seyrig C and Holowacz S provided essential materials (bacterial strains and the probiotic supplement Lactibiane Tolerance®); Nébot-Vivinus M, Bzioueche H, Piche T, Theodorou V analyzed the data or performed statistical analyses.
Supported by PiLeJe, 37, quai-de-Grenelle, bâtiment Pollux, 75738 Paris cedex 15
Correspondence to: Thierry Piche, Professor, Department of Gastroenterology, CHU de Nice, Hôpital Archet 2, 06103 Nice, France. piche.t@chu-nice.fr
Telephone: +33-4-92036399  Fax: + 33-4-92036075
Received: November 29, 2013
Revised: February 18, 2014
Accepted: March 4, 2014
Published online: June 14, 2014
Processing time: 198 Days and 20.5 Hours
Abstract

AIM: To investigate the effect of the probiotic combination Lactibiane Tolerance® (LT) on epithelial barrier function in vitro and in vivo.

METHODS: The effect of the multispecies probiotic LT was assessed on several models of epithelial barrier function both in vitro (in basal and inflammatory conditions) and in vivo [visceral hypersensitivity induced by chronic stress or by colonic perfusion of a fecal supernatant (FSN) from patients with irritable bowel syndrome (IBS)]. In vitro, we measured the permeability of confluent T84 cell monolayers incubated with or without LT by evaluating the paracellular flux of macromolecules, in basal conditions and after stimulation with lipopolysaccharide (LPS) or with conditioned medium of colonic biopsies from IBS patients (IBS-CM). In vivo, male C57/Bl6 mice received orally NaCl or LT for 15 d and were submitted to water avoidance stress (WAS) before evaluating visceral sensitivity by measuring the myoelectrical activity of the abdominal muscle and the paracellular permeability with 51Cr-EDTA. Permeability and sensitivity were also measured after colonic instillation of FSN. Tight-junctions were assessed by immunoblotting and TLR-4 expression was evaluated by immunohistochemistry

RESULTS: Incubation of T84 cell monolayers with LT in basal conditions had no significant effect on permeability (P > 0.05 vs culture medium). By contrast, addition of LT bacterial bodies (LT) completely prevented the LPS-induced increase in paracellular permeability (P < 0.01 vs LPS 10 ng/mL (LPS 10); P < 0.01 vs LPS 100 ng/mL (LPS 100), P > 0.05 vs culture medium). The effect was dose dependent as addition of 109 LT bacterial bodies induced a stronger decrease in absorbance than 106 LT (109 LT + LPS 10: -20.1% ± 13.4, P < 0.01 vs LPS 10; 106 LT + LPS 10: -11.6% ± 6.2, P < 0.01 vs LPS 10; 109 LT + LPS 100: -14.4% ± 5.5, P < 0.01 vs LPS 100; 106 LT + LPS 100: -11.6% ± 7.3, P < 0.05 vs LPS 100). Moreover, the increase in paracellular permeability induced by culturing T84 cells with conditioned medium of colonic biopsies from IBS patients (IBS-CM) was completely inhibited in the presence of 109 LT (P < 0.01 vs IBS-CM). LT also significantly prevented the epithelial disruption induced by intracolonic infusion of fecal supernatant from IBS patients (P < 0.01 vs IBS FSN) or water avoidance stress P < 0.01 vs WAS) in C57/Bl6 mice and increased the expression of occludin in vitro and in vivo, as assessed by immnunoblotting. The WAS-induced effect on visceral sensitivity was prevented by LT treatment since values obtained for all steps of colorectal distension were significantly (P < 0.01) different from the WAS group. Finally, LT down-regulated the response mediated through TLR-4 in vitro (decrease in tumor necrosis factor α secretion in response to LPS: -65.8% for 109 LT and -52.5% for 106 LT, P < 0.01 vs LPS) and in vivo (inhibition of WAS induced an increase in TLR-4 expression in the LT treated mice colon, P < 0.01 vs WAS).

CONCLUSION: The probiotic LT mix prevented the disruption to the epithelial barrier induced by LPS, stress or colonic soluble factors from IBS patients and prevented visceral hypersensitivity.

Keywords: Irritable bowel syndrome; Probiotic; Intestinal epithelial barrier permeability; Hypersensitivity

Core tip: Some probiotics are effective in treatment of irritable bowel syndrome (IBS). The pathophysiology of IBS involves disruption of the epithelial barrier together with low grade inflammation, which represent peripheral targets for probiotics. This study aimed to elucidate the effect of a multispecies combination of bacterial strains on in vitro and in vivo models mimicking IBS. This commercialized combination was able to prevent visceral hypersensitivity and to repair the disruption to the epithelial barrier induced by lipopolysaccharide, stress or colonic soluble factors from IBS patients.