title: Inhibitory effects of Lactobacillus casei subsp. rhamnosus on Salmonella lipopolysaccharide-induced inflammation and epithelial barrier dysfunction in a co-culture model using Caco-2/peripheral blood mononuclear cells.
abstract: In this study, we investigated the anti-inflammatory and reinforcing barrier effects of Lactobacillus casei subsp. rhamnosus (Lcr35) on Caco-2 intestinal epithelial cells already exposed to Salmonella LPS. Using the Transwell co-culture model, Salmonella LPS was apically added to polarized Caco-2 cells co-cultured with peripheral blood mononuclear cells (PBMCs) in the basolateral compartment. LPS-stimulated Caco-2 cells were incubated with Lcr35 for 1, 6, 24 or 48 h. Apical inoculation of Lcr35 after 48 h significantly inhibited the basolateral secretion of interleukin-8 (IL-8) in the Caco-2/PBMC co-culture. The PCR analysis showed that Lcr35 significantly downregulated mRNA expression of monocyte chemoattractant protein 1 (MCP-1) (P<0.05) and had a trend of decreasing mRNA expression of IL-8 (P=0.05), but did not alter mRNA expression of transforming growth factor-beta1 in LPS-stimulated Caco-2 cells at 48 h after addition of Lcr35. Compared to non-LPS-pretreated controls, transepithelial electrical resistance (TEER) of the polarized Caco-2 cell monolayers pretreated with LPS for 48 h was decreased by 9.9 % (P<0.05). Additionally, compared to those cells only treated with LPS, apical co-incubation with Lcr35 showed biphasic TEER levels increased by 12.1 % (P<0.001), 5.7 % (P<0.05) and 86.8 % (P<0.001) in the Caco-2 cell monolayers compared to those without Lcr35 treatment after 1, 6 and 48 h, respectively. In conclusion, Lcr35 can exert anti-inflammatory effects and ameliorate barrier dysfunction in the Salmonella LPS-pretreated inflamed intestinal epithelium in vitro.
authors.affiliation: Division of Medical Engineering Research, National Health Research Institutes, Miaoli, Taiwan, ROC.|Department of Chemical Engineering and Biotechnology, National Taipei University Technology, Taipei, Taiwan, ROC.
authors.forename: Hsu-Wei
authors.initials: HW
authors.lastname: Fang
authors.affiliation: Centre for Paediatric Gastroenterology, Royal Free Campus, University College London Medical School, London, UK.
authors.forename: Shiuh-Bin
authors.initials: SB
authors.lastname: Fang
authors.affiliation: Department of Medical Research, Mackay Memorial Hospital, Taipei, Taiwan, ROC.
authors.forename: Jen-Shiu
authors.initials: JS
authors.lastname: Chiang Chiau
authors.affiliation: Mackay Medicine, Nursing and Management College, Taipei, Taiwan, ROC.|Department of Paediatrics, Mackay Memorial Hospital, Taipei, Taiwan, ROC.
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Yeung
authors.affiliation: Department of Paediatrics, Mackay Memorial Hospital, Taipei, Taiwan, ROC.
authors.forename: Wai-Tao
authors.initials: WT
authors.lastname: Chan
authors.affiliation: Department of Paediatrics, Mackay Memorial Hospital, Taipei, Taiwan, ROC.
authors.forename: Chuen-Bin
authors.initials: CB
authors.lastname: Jiang
authors.affiliation: Department of Medical Research, Mackay Memorial Hospital, Taipei, Taiwan, ROC.
authors.forename: Mei-Lien
authors.initials: ML
authors.lastname: Cheng
authors.affiliation: Department of Paediatrics, Taipei Medical University, Taipei, Taiwan, ROC.|Department of Paediatrics, Mackay Memorial Hospital, Taipei, Taiwan, ROC.
authors.forename: Hung-Chang
authors.initials: HC
authors.lastname: Lee
mesh_terms: D018938:Caco-2 Cells; D018932:Chemokine CCL2 / Q000096:biosynthesis; D018920:Coculture Techniques; D015536:Down-Regulation; D017097:Electric Impedance; D004847:Epithelial Cells / Q000187:drug effects / Q000276:immunology* / Q000382:microbiology*; D015870:Gene Expression; D006801:Humans; D007249:Inflammation; D016209:Interleukin-8 / Q000378:metabolism; D007780:Lacticaseibacillus casei / Q000502:physiology*; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology / Q000302:isolation & purification / Q000633:toxicity*; D012475:Salmonella / Q000737:chemistry*; D053773:Transforming Growth Factor beta1 / Q000096:biosynthesis
chemical_list: C111835:CCL2 protein, human; D018932:Chemokine CCL2; D016209:Interleukin-8; D008070:Lipopolysaccharides; D053773:Transforming Growth Factor beta1