title: Effects of Lactobacillus johnsonii and Lactobacillus reuteri on gut barrier function and heat shock proteins in intestinal porcine epithelial cells.
abstract: Heat shock proteins (HSPs) are a set of highly conserved proteins that can serve as intestinal gate keepers in gut homeostasis. Here, effects of a probiotic, Lactobacillus rhamnosus GG (LGG), and two novel porcine isolates, Lactobacillus johnsonii strain P47-HY and Lactobacillus reuteri strain P43-HUV, on cytoprotective HSP expression and gut barrier function, were investigated in a porcine IPEC-J2 intestinal epithelial cell line model. The IPEC-J2 cells polarized on a permeable filter exhibited villus-like cell phenotype with development of apical microvilli. Western blot analysis detected HSP expression in IPEC-J2 and revealed that L. johnsonii and L. reuteri strains were able to significantly induce HSP27, despite high basal expression in IPEC-J2, whereas LGG did not. For HSP72, only the supernatant of L. reuteri induced the expression, which was comparable to the heat shock treatment, which indicated that HSP72 expression was more stimulus specific. The protective effect of lactobacilli was further studied in IPEC-J2 under an enterotoxigenic Escherichia coli (ETEC) challenge. ETEC caused intestinal barrier destruction, as reflected by loss of cell-cell contact, reduced IPEC-J2 cell viability and transepithelial electrical resistance, and disruption of tight junction protein zonula occludens-1. In contrast, the L. reuteri treatment substantially counteracted these detrimental effects and preserved the barrier function. L. johnsonii and LGG also achieved barrier protection, partly by directly inhibiting ETEC attachment. Together, the results indicate that specific strains of Lactobacillus can enhance gut barrier function through cytoprotective HSP induction and fortify the cell protection against ETEC challenge through tight junction protein modulation and direct interaction with pathogens.
authors.affiliation: Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, Uppsala, Sweden Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden.
authors.forename: Hao-Yu
authors.initials: HY
authors.lastname: Liu
authors.affiliation: Department of Microbiology, Uppsala BioCenter, Swedish University of Agricultural Sciences, Uppsala, Sweden.
authors.forename: Stefan
authors.initials: S
authors.lastname: Roos
authors.affiliation: Department of Microbiology, Uppsala BioCenter, Swedish University of Agricultural Sciences, Uppsala, Sweden.
authors.forename: Hans
authors.initials: H
authors.lastname: Jonsson
authors.affiliation: Department of Medical Cell Biology, Uppsala University, Uppsala, Sweden.
authors.forename: David
authors.initials: D
authors.lastname: Ahl
authors.affiliation: Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, Uppsala, Sweden.
authors.forename: Johan
authors.initials: J
authors.lastname: Dicksved
authors.affiliation: Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, Uppsala, Sweden.
authors.forename: Jan Erik
authors.initials: JE
authors.lastname: Lindberg
authors.affiliation: Department of Animal Nutrition and Management, Swedish University of Agricultural Sciences, Uppsala, Sweden torbjorn.lundh@slu.se.
authors.forename: Torbjörn
authors.initials: T
authors.lastname: Lundh
keywords: Cytoprotective heat shock protein 27 and 72; IPEC‐J2 cell line; intestinal barrier integrity; probiotic lactobacilli; tight junction protein
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