title: Lactobacillus plantarum inhibits intestinal epithelial barrier dysfunction induced by unconjugated bilirubin.
abstract: Although a large number of in vitro and in vivo tests have confirmed that taking probiotics can improve the intestinal barrier, few studies have focused on the relationship between probiotics and the intestinal epithelial barrier in hyperbilirubinaemia. To investigate the effects of and mechanisms associated with probiotic bacteria (Lactobacillus plantarum; LP) and unconjugated bilirubin (UCB) on the intestinal epithelial barrier, we measured the viability, apoptotic ratio and protein kinase C (PKC) activity of Caco-2 cells. We also determined the distribution and expression of tight junction proteins such as occludin, zonula occludens (ZO)-1, claudin-1, claudin-4, junctional adhesion molecule (JAM)-1 and F-actin using confocal laser scanning microscopy, immunohistochemistry, Western blotting and real-time quantitative PCR. The present study demonstrated that high concentrations of UCB caused obvious cytotoxicity and decreased the transepithelial electrical resistance (TER) of the Caco-2 cell monolayer. Low concentrations of UCB inhibited the expression of tight junction proteins and PKC but could induce UDP-glucuronosyltransferases 1 family-polypeptide A1 (UGT1A1) expression. UCB alone caused decreased PKC activity, serine phosphorylated occludin and ZO-1 levels. After treatment with LP, the effects of UCB on TER and apoptosis were mitigated; LP also prevented aberrant expression and rearrangement of tight junction proteins. Moreover, PKC activity and serine phosphorylated tight junction protein levels were partially restored after treatment with LP, LP exerted a protective effect against UCB damage to Caco-2 monolayer cells, and it restored the structure and distribution of tight junction proteins by activating the PKC pathway. In addition, UGT1A1 expression induced by UCB in Caco-2 cells could ameliorate the cytotoxicity of UCB.
authors.affiliation: Shanghai Jiaotong University Affiliated Sixth People's Hospital, 600 Yishan Road, Shanghai, China.
authors.forename: Yukun
authors.initials: Y
authors.lastname: Zhou
authors.forename: Huanlong
authors.initials: H
authors.lastname: Qin
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
authors.forename: Tongyi
authors.initials: T
authors.lastname: Shen
authors.forename: Hongqi
authors.initials: H
authors.lastname: Chen
authors.forename: Yanlei
authors.initials: Y
authors.lastname: Ma
authors.forename: Zhaoxin
authors.initials: Z
authors.lastname: Chu
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Liu
mesh_terms: D017209:Apoptosis; D001663:Bilirubin / Q000378:metabolism*; D018938:Caco-2 Cells; D002470:Cell Survival; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology; D005765:Gastrointestinal Agents / Q000627:therapeutic use; D014453:Glucuronosyltransferase / Q000378:metabolism; D006801:Humans; D006932:Hyperbilirubinemia / Q000188:drug therapy* / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism / Q000382:microbiology; D048191:Lactiplantibacillus plantarum; D008565:Membrane Proteins / Q000378:metabolism*; D062793:Occludin; D010766:Phosphorylation; D019936:Probiotics / Q000627:therapeutic use*; D011493:Protein Kinase C / Q000378:metabolism; D012694:Serine / Q000378:metabolism; D019108:Tight Junctions / Q000378:metabolism*
chemical_list: D005765:Gastrointestinal Agents; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; D012694:Serine; D014453:Glucuronosyltransferase; D011493:Protein Kinase C; D001663:Bilirubin