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.
J Med Microbiol. 2010 May;59(Pt 5):573-579. doi: 10.1099/jmm.0.009662-0. Epub 2010 Jan 28.
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.
在这项研究中，我们研究了干酪乳杆菌亚种。鼠李糖乳杆菌（Lcr35）对已经暴露于沙门氏菌 LPS 的 Caco-2 肠上皮细胞的抗炎和增强屏障作用。使用 Transwell 共培养模型，将沙门氏菌 LPS apical 加至与外周血单核细胞（PBMCs）共培养的极化 Caco-2 细胞的基底外侧。将 LPS 刺激的 Caco-2 细胞与 Lcr35 孵育 1、6、24 或 48 h。在 48 h 后 apical 接种 Lcr35 可显著抑制 Caco-2/PBMC 共培养物中基底外侧白细胞介素-8（IL-8）的分泌。PCR 分析表明，Lcr35 显著下调单核细胞趋化蛋白 1（MCP-1）的 mRNA 表达（P<0.05），并且具有降低 IL-8 的 mRNA 表达的趋势（P=0.05），但在加入 Lcr35 48 h 后，对 LPS 刺激的 Caco-2 细胞中转化生长因子-β1 的 mRNA 表达没有改变。与未经 LPS 预处理的对照相比，经 LPS 预处理 48 h 的极化 Caco-2 细胞单层的跨上皮电阻（TEER）降低了 9.9％（P<0.05）。此外，与仅用 LPS 处理的细胞相比，用 Lcr35 apical 共孵育可使 TEER 水平分别增加 12.1％（P<0.001），5.7％（P<0.05）和 86.8％（P<0.001），在无 Lcr35 处理的 Caco-2 细胞单层中，分别在 1、6 和 48 h 后。总之，Lcr35 可在体外发挥抗炎作用，并改善沙门氏菌 LPS 预处理的炎症性肠上皮的屏障功能障碍。
