Department of Cell and Molecular Biology of Drugs, Faculty of Pharmacy, Comenius University in Bratislava, Odbojárov 10, Bratislava, Slovak Republic.
Antonie Van Leeuwenhoek. 2020 Aug;113(8):1191-1200. doi: 10.1007/s10482-020-01426-1. Epub 2020 May 14.
Adhesion of probiotic bacteria to the mucus layer lining the gastrointestinal tract is necessary for its effective colonisation and specific therapeutic effects. Enrichment of growth medium with mucin might stimulate bacterial adhesion, probably by increasing the expression of surface structures responsible for bacteria-gut epithelia and/or mucus interactions. The aim of this study was to determine if pre-cultivation of potentially probiotic strain Lactobacillus reuteri E (LRE) with mucin stimulates its adherence to colon cell line HT-29 and if the increased adhesion modulates mucin expression in these cells. The mucin-producing HT-29 cell line was co-cultivated for 2 h with LRE grown in MRS broth or MRS broth enriched with pig gastric mucin (LRE + M). The adherence ability of LRE was evaluated microscopically and by plate counting. The relative gene expression was measured by qPCR. Pre-cultivation of LRE in mucin enriched medium significantly increased its adhesion to 14 days HT-29 in comparison with LRE by both methods (28.64% vs. 23.83%, evaluated microscopically, and 14.31 ± 3.95 × 10 CFU ml vs. 8.54 ± 0.43 × 10 CFU ml, evaluated by plate counting). MUC2, MUC5AC, and IL-10 were significantly upregulated after co-cultivation with LRE + M in comparison to LRE and control group (lactobacilli-free HT-29). Obtained results suggest that pre-cultivation of lactobacilli with mucin may not only stimulate their adhesion abilities but also promote their effectiveness to modulate the pathways involved in the pathophysiology of some diseases, e.g., with defective mucin synthesis in ulcerative colitis or colorectal cancer.
益生菌黏附在胃肠道黏膜层是其有效定植和发挥特定治疗效果的必要条件。在生长培养基中添加黏蛋白可能会刺激细菌黏附,这可能是通过增加负责细菌-肠道上皮和/或黏液相互作用的表面结构的表达来实现的。本研究旨在确定预先培养潜在益生菌菌株罗伊氏乳杆菌 E(LRE)与黏蛋白是否会刺激其黏附到结肠细胞系 HT-29,以及增加的黏附是否会调节这些细胞中的黏蛋白表达。用 MRS 肉汤或富含猪胃黏蛋白的 MRS 肉汤(LRE+M)培养 LRE 2 小时,与 HT-29 细胞系共培养。通过显微镜和平板计数评估 LRE 的黏附能力。通过 qPCR 测量相对基因表达。与 LRE 相比,LRE 在富含黏蛋白的培养基中预培养可显著提高其对 14 天 HT-29 的黏附能力(通过显微镜评估为 28.64%对 23.83%,通过平板计数评估为 14.31±3.95×10 CFU ml 对 8.54±0.43×10 CFU ml)。与 LRE 和对照组(不含乳酸菌的 HT-29)相比,LRE+M 共培养后 MUC2、MUC5AC 和 IL-10 的表达显著上调。这些结果表明,预先培养黏蛋白可能不仅可以刺激乳酸菌的黏附能力,还可以增强其调节某些疾病发病机制相关途径的有效性,例如溃疡性结肠炎或结直肠癌中黏蛋白合成缺陷。