Jakubczyk Dominika, Leszczyńska Katarzyna, Pacyga-Prus Katarzyna, Kozakiewicz Dominika, Kazana-Płuszka Wioletta, Gełej Dominika, Migdał Paweł, Kruszakin Roksana, Zabłocka Agnieszka, Górska Sabina
Laboratory of Microbiome Immunobiology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland.
Inter-Departmental Laboratory of Instrumental Analysis and Preparation, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland.
BMC Microbiol. 2024 Feb 19;24(1):60. doi: 10.1186/s12866-023-03179-z.
The impact of probiotic strains on host health is widely known. The available studies on the interaction between bacteria and the host are focused on the changes induced by bacteria in the host mainly. The studies determining the changes that occurred in the bacteria cells are in the minority. Within this paper, we determined what happens to the selected Bifidobacterium adolescentis and Bifidobacterium longum ssp. longum in an experimental environment with the intestinal epithelial layer. For this purpose, we tested the bacteria cells' viability, redox activity, membrane potential and enzymatic activity in different environments, including CaCo-2/HT-29 co-culture, cell culture medium, presence of inflammatory inductor (TNF-α) and oxygen.
We indicated that the external milieu impacts the viability and vitality of bacteria. Bifidobacterium adolescentis decrease the size of the live population in the cell culture medium with and without TNF-α (p < 0.001 and p < 0.01 respectively). In contrast, Bifidobacterium longum ssp. longum significantly increased survivability in contact with the eukaryotic cells and cell culture medium (p < 0.001). Bifidobacterium adolescentis showed significant changes in membrane potential, which was decreased in the presence of eukaryotic cells (p < 0.01), eukaryotic cells in an inflammatory state (p < 0.01), cell culture medium (p < 0.01) and cell culture medium with TNF-α (p < 0.05). In contrast, Bifidobacterium longum ssp. longum did not modulate membrane potential. Instead, bacteria significantly decreased the redox activity in response to milieus such as eukaryotic cells presence, inflamed eukaryotic cells as well as the culture medium (p < 0.001). The redox activity was significantly different in the cells culture medium vs the presence of eukaryotic cells (p < 0.001). The ability to β-galactosidase production was different for selected strains: Bifidobacterium longum ssp. longum indicated 91.5% of positive cells, whereas Bifidobacterium adolescentis 4.34% only. Both strains significantly reduced the enzyme production in contact with the eukaryotic milieu but not in the cell culture media.
The environmental-induced changes may shape the probiotic properties of bacterial strains. It seems that the knowledge of the sensitivity of bacteria to the external environment may help to select the most promising probiotic strains, reduce research costs, and contribute to greater reproducibility of the obtained probiotic effects.
益生菌菌株对宿主健康的影响广为人知。现有的关于细菌与宿主相互作用的研究主要集中在细菌在宿主中引起的变化上。确定细菌细胞中发生的变化的研究占少数。在本文中,我们确定了在具有肠上皮层的实验环境中,选定的青春双歧杆菌和长双歧杆菌亚种会发生什么。为此,我们在不同环境中测试了细菌细胞的活力、氧化还原活性、膜电位和酶活性,包括CaCo-2/HT-29共培养、细胞培养基、存在炎性诱导剂(TNF-α)和氧气的环境。
我们表明外部环境会影响细菌的活力和生命力。青春双歧杆菌在有和没有TNF-α的细胞培养基中都会减少活菌数量(分别为p < 0.001和p < 0.01)。相比之下,长双歧杆菌亚种在与真核细胞和细胞培养基接触时显著提高了存活率(p < 0.001)。青春双歧杆菌的膜电位有显著变化,在存在真核细胞(p < 0.01)、处于炎症状态的真核细胞(p < 0.01)、细胞培养基(p < 0.01)和含有TNF-α的细胞培养基(p < 0.05)时膜电位降低。相比之下,长双歧杆菌亚种没有调节膜电位。相反,细菌在诸如存在真核细胞、发炎的真核细胞以及培养基等环境中会显著降低氧化还原活性(p < 0.001)。细胞培养基中的氧化还原活性与存在真核细胞时的氧化还原活性有显著差异(p < 0.001)。选定菌株的β-半乳糖苷酶产生能力不同:长双歧杆菌亚种显示91.5%的阳性细胞,而青春双歧杆菌仅为4.34%。两种菌株在与真核环境接触时都会显著降低酶的产生,但在细胞培养基中则不会。
环境诱导的变化可能会塑造细菌菌株的益生菌特性。似乎了解细菌对外部环境的敏感性可能有助于选择最有前景的益生菌菌株,降低研究成本,并有助于提高所获得的益生菌效果的重现性。