Mucin pre-cultivated Lactobacillus reuteri E shows enhanced adhesion and increases mucin expression in HT-29 cells

• Published in: Antonie van Leeuwenhoek 2020-08, Vol.113 (8), p.1191-1200
• Main Authors: Dudík, Boris, Kiňová Sepová, Hana, Bilka, František, Pašková, Ľudmila, Bilková, Andrea
• Format: Article
• Language: English
• Subjects: Adhesion; Animals; Bacteria; Biomedical and Life Sciences; Biotechnology; Cell Adhesion; Cell adhesion & migration; Colon; Colorectal carcinoma; Cultivation; Enrichment; Epithelial Cells - microbiology; Evaluation; Gastrointestinal system; Gastrointestinal tract; Gene expression; HT29 Cells - metabolism; Humans; Interleukin 10; Intestinal Mucosa - metabolism; Lactobacilli; Lactobacillus; Lactobacillus reuteri; Lactobacillus reuteri - genetics; Lactobacillus reuteri - metabolism; Life Sciences; Medical Microbiology; Microbiology; Mucin; Mucins - metabolism; Mucus; Original Paper; Pathophysiology; Plant Sciences; Probiotics; Soil Science & Conservation; Swine; Ulcerative colitis
• ISSN: 0003-6072; 1572-9699
• DOI: 10.1007/s10482-020-01426-1

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 6  CFU ml −1 vs. 8.54 ± 0.43 × 10 6  CFU ml −1 , 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.