title: Lactobacillus plantarum MB452 enhances the function of the intestinal barrier by increasing the expression levels of genes involved in tight junction formation.
abstract: BACKGROUND
  Intestinal barrier function is important for preserving health, as a compromised barrier allows antigen entry and can induce inflammatory diseases. Probiotic bacteria can play a role in enhancing intestinal barrier function; however, the mechanisms are not fully understood. Existing studies have focused on the ability of probiotics to prevent alterations to tight junctions in disease models, and have been restricted to a few tight junction bridging proteins. No studies have previously investigated the effect of probiotic bacteria on healthy intestinal epithelial cell genes involved in the whole tight junction signalling pathway, including those encoding for bridging, plaque and dual location tight junction proteins. Alteration of tight junction signalling in healthy humans is a potential mechanism that could lead to the strengthening of the intestinal barrier, resulting in limiting the ability of antigens to enter the body and potentially triggering undesirable immune responses.
  
  
  RESULTS
  The effect of Lactobacillus plantarum MB452 on tight junction integrity was determined by measuring trans-epithelial electrical resistance (TEER) across Caco-2 cell layers. L. plantarum MB452 caused a dose-dependent TEER increase across Caco-2 cell monolayers compared to control medium. Gene expression was compared in Caco-2 cells untreated or treated with L. plantarum MB452 for 10 hours. Caco-2 cell RNA was hybridised to human oligonucleotide arrays. Data was analysed using linear models and differently expressed genes were examined using pathway analysis tools. Nineteen tight junction-related genes had altered expression levels in response to L. plantarum MB452 (modified-P < 0.05, fold-change > 1.2), including those encoding occludin and its associated plaque proteins that anchor it to the cytoskeleton. L. plantarum MB452 also caused changes in tubulin and proteasome gene expression levels which may be linked to intestinal barrier function. Caco-2 tight junctions were visualised by fluorescent microscopy of immuno-stained occludin, zona occludens (ZO)-1, ZO-2 and cingulin. Caco-2 cells treated with L. plantarum MB452 had higher intensity fluorescence of each of the four tight junction proteins compared to untreated controls.
  
  
  CONCLUSIONS
  This research indicates that enhancing the expression of genes involved in tight junction signalling is a possible mechanism by which L. plantarum MB452 improves intestinal barrier function.
authors.affiliation: AgriFoods & Health Section, Food & Textiles Group, AgResearch Grasslands, Private Bag 11008, Palmerston North 4442, New Zealand. rachel.anderson@agresearch.co.nz
authors.forename: Rachel C
authors.initials: RC
authors.lastname: Anderson
authors.forename: Adrian L
authors.initials: AL
authors.lastname: Cookson
authors.forename: Warren C
authors.initials: WC
authors.lastname: McNabb
authors.forename: Zaneta
authors.initials: Z
authors.lastname: Park
authors.forename: Mark J
authors.initials: MJ
authors.lastname: McCann
authors.forename: William J
authors.initials: WJ
authors.lastname: Kelly
authors.forename: Nicole C
authors.initials: NC
authors.lastname: Roy
mesh_terms: D018938:Caco-2 Cells; D017097:Electric Impedance; D004847:Epithelial Cells / Q000737:chemistry / Q000382:microbiology / Q000502:physiology; D015870:Gene Expression; D006801:Humans; D007422:Intestines / Q000737:chemistry / Q000166:cytology / Q000382:microbiology / Q000502:physiology*; D048191:Lactiplantibacillus plantarum / Q000502:physiology*; D015398:Signal Transduction; D019108:Tight Junctions / Q000737:chemistry / Q000378:metabolism* / Q000382:microbiology