食品与发酵工业2024，Vol.50Issue(24)：191-200,10.DOI:10.13995/j.cnki.11-1802/ts.038094
鼠李糖乳酪杆菌KF7发酵乳上清对脂多糖诱导的Caco-2细胞单层屏障损伤的保护作用及其机制研究
Study on protective effects and mechanism of Lacticaseibacillus rhamnosus KF7 fermented milk supernatant on lipopolysaccharide-induced epithelial barrier damage in Caco-2 cell monolayer model
摘要
Abstract
Damage to the intestinal barrier is associated with many diseases.Preventing and improving impaired intestinal barrier function through dietary interventions,especially probiotics,may be a promising therapeutic strategy.This study utilized lipopolysaccharide to induce damage in the Caco-2 cell monolayer barrier model to investigate the protective effect of Lacticaseibacillus rhamnosus KF7 fer-mented milk supernatant on the intestinal barrier in vitro.Cell Counting Kit-8(CCK-8)assay was used to detect the effects of KF7 fermen-ted milk supernatant on cell viability.A resistivity meter and fluorescein isothiocyanate-dextrose were used to examine the structural integri-ty and permeability of the intestinal epithelial barrier.Fluorescent probe and colorimetric assay were used to detect the intracellular levels of reactive oxygen species and malondialdehyde(MDA),and qRT-PCR was used to analyze the gene expression levels of tight junction proteins,pro-inflammatory cytokines,anti-inflammatory cytokines,and markers of oxidative stress and inflammation-related pathways.Re-sults revealed that 1％-5％Lacticaseibacillus rhamnosus KF7 fermented milk supernatant did not impair Caco-2 cell viability,but protec-ted the Caco-2 cell monolayer barrier,decreasing lipopolysaccharides-induced inflammatory response(down-regulation of gene expression of TNF-a,IL-1β,and IL-6,and up-regulation of IL-10)and oxidative stress(reduction of MDA content),increasing the expression of intercellular tight junction protein-related genes(Claudin-1 and Occludin),improving cell monolayer trans-epithelial electrical resistance,and reducing the permeability,thus recovered the integrity of Caco-2 cell monolayer barrier.Mechanistically,Lacticaseibacillus rhamnosus KF7 fermented milk supernatant may antagonize oxidative stress by promoting the Nrf2/NQO1 signaling pathway,and decrease the inflam-matory response and intestinal epithelial inter-cellular permeability by inhibiting the TLR/MyD88/NF-κB signaling pathway and down-stream myosin lightchain kinase(MLCK)gene expression.The results of this study may provide some theoretical basis for Lacticaseibacillus rhamnosus KF7 fermented dairy products as a promising functional probiotic dietary supplement for protecting intestinal health.关键词
鼠李糖乳酪杆菌KF7/Caco-2细胞/跨上皮电阻/肠上皮屏障/氧化应激/炎症反应Key words
Lacticaseibacillus rhamnosus KF7/Caco-2 cells/transepithelial electrical resistance/intestinal epithelial barrier/oxidative stress/inflammatory response引用本文复制引用
董奇奇,张怡琳,游春苹..鼠李糖乳酪杆菌KF7发酵乳上清对脂多糖诱导的Caco-2细胞单层屏障损伤的保护作用及其机制研究[J].食品与发酵工业,2024,50(24):191-200,10.基金项目
上海市优秀技术带头人计划(20XD1430100) （20XD1430100）
市国资委能力提升项目(2022013) （2022013）
