Abstract
BackgroundPrevious studies indicated that the micro integral membrane protein located within the media place of the integral membrane protein of Lactobacillus plantarum CGMCC 1258 had protective effects against the intestinal epithelial injury. In our study, we mean to establish micro integral membrane protein -knockout Lactobacillus plantarum (LPKM) to investigate the change of its protective effects and verify the role of micro integral membrane protein on protection of normal intestinal barrier function.MethodsBinding assay and intestinal permeability were performed to verify the protective effects of micro integral membrane protein on intestinal permeability in vitro and in vivo. Molecular mechanism was also determined as the zonulin pathway. Clinical data were also collected for further verification of relationship between zonulin level and postoperative septicemia.ResultsLPKM got decreased inhibition of EPEC adhesion to NCM460 cells. LPKM had lower ability to alleviate the decrease of intestinal permeability induced by enteropathogenic-e.coli, and prevent enteropathogenic-e.coli -induced increase of zonulin expression. Overexpression of zonulin lowered the intestinal permeability regulated by Lactobacillus plantarum. There was a positive correlation between zonulin level and postoperative septicemia. Therefore, micro integral membrane protein could be necessary for the protective effects of Lactobacillus plantarum on intestinal barrier.ConclusionMIMP might be a positive factor for Lactobacillus plantarum to protect the intestinal epithelial cells from injury, which could be related to the zonulin pathway.
Highlights
Previous studies indicated that the micro integral membrane protein located within the media place of the integral membrane protein of Lactobacillus plantarum CGMCC 1258 had protective effects against the intestinal epithelial injury
PCR amplicons over the IMP region indicated that compared with Lactobacillus plantarum (LP) of near 1000 bp, LPKM lost about 200 bp in the genes surrounding the deletion of LPKM (Figure 1A)
Detection of EPEC adherence indicated that the adhesion rate of EPEC to NCM460 cells was reduced significantly when LP was added, while adding LPKM had no effects on the inhibition of EPEC adhesion
Summary
Previous studies indicated that the micro integral membrane protein located within the media place of the integral membrane protein of Lactobacillus plantarum CGMCC 1258 had protective effects against the intestinal epithelial injury. We mean to establish micro integral membrane protein -knockout Lactobacillus plantarum (LPKM) to investigate the change of its protective effects and verify the role of micro integral membrane protein on protection of normal intestinal barrier function. The lactobacillus surface layer protein (SLP) has been raised as a key component mediating the protection conferred by LP to intestinal epithelial cells (IECs) [7,8]. Soluble factors, including p75 and p40, extracted from Lactobacillus rhamnosus GG culture broth supernatant, showed protective effects on IECs, which was mediated by Akt pathway [13,14]. A few studies have, investigated the structure and functions of SLP [8,10,15,16,17,18], due to the specific hydrophilic and hydrophobic properties and technical difficulties associated with SLP purification, limiting the investigation of SLP binding domains
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