Department of Korean Medicine, Dongguk University, Goyang, Republic of Korea.
PLoS One. 2020 Mar 9;15(3):e0229647. doi: 10.1371/journal.pone.0229647. eCollection 2020.
Probiotic bacteria have the ability to modulate host immune responses and have potent therapeutic functional effects against several diseases, including inflammatory diseases. However, beneficial effects of probiotics are strain specific and their interactions with host immune cells to modulate inflammatory response are largely unknown. Intestinal epithelial cells (IECs), which are the first line of defense against invading pathogens, and connects between commensals/probiotics and immune system; therefore, in this study, we used human IECs to assess the probiotic effects of three selected Lactobacillus strains in vitro. An HT-29 colonic epithelial cell and HT-29/blood mononuclear cells co-culture system were stimulated with Lactobacillus followed by Salmonella for different hours, after which the mRNA level of cytokines, β-defensin-2 and negative regulators for TLR signaling and protein levels of ZO-1 and IκB-α were analyzed by real-time polymerase chain reaction and western blot analysis. L. brevis decreased Salmonella induced IL-6, IL-8, MCP-1 and IL-1β levels, whereas L. pentosus suppressed IL-6 and MCP-1 in HT-29 cells. Moreover, L. brevis was able to increase the mRNA levels of A20, Tollip, SIGIRR and IRAKM, while L. pentosus reduced the levels of A20, and IRAKM in response to Salmonella. In addition, decrease in protein level of TNF-α and increase in mRNA level of IL-10 was observed in L. brevis and L. pentosus treated HT-29 cells. Lactobacillus strains were differentially modulated ZO-1 and p-IκB-α in HT-29 cells treated with Salmonella. Overall, the results of this study indicate that Lactobacillus strains attenuate Salmonella induced inflammatory responses through beneficial modulation of TLR negative regulators and the NF-κB pathway.
益生菌具有调节宿主免疫反应的能力,并对多种疾病具有有效的治疗功能,包括炎症性疾病。然而,益生菌的有益作用是菌株特异性的,其与宿主免疫细胞相互作用以调节炎症反应在很大程度上尚不清楚。肠上皮细胞(IECs)是抵御入侵病原体的第一道防线,连接共生菌/益生菌和免疫系统;因此,在这项研究中,我们使用人肠上皮细胞在体外评估了三种选定的乳杆菌菌株的益生菌作用。使用 HT-29 结肠上皮细胞和 HT-29/血液单核细胞共培养系统,用乳杆菌刺激后再用沙门氏菌刺激不同时间,然后通过实时聚合酶链反应和 Western blot 分析测定细胞因子、β-防御素-2 和 TLR 信号的负调节剂以及 ZO-1 和 IκB-α 的蛋白水平。L. brevis 降低了沙门氏菌诱导的 IL-6、IL-8、MCP-1 和 IL-1β 水平,而 L. pentosus 抑制了 HT-29 细胞中的 IL-6 和 MCP-1。此外,L. brevis 能够增加 A20、Tollip、SIGIRR 和 IRAKM 的 mRNA 水平,而 L. pentosus 降低了 A20 和 IRAKM 的水平,以响应沙门氏菌。此外,在 L. brevis 和 L. pentosus 处理的 HT-29 细胞中观察到 TNF-α 蛋白水平降低和 IL-10 mRNA 水平增加。乳杆菌菌株通过有益地调节 TLR 负调节剂和 NF-κB 途径来调节沙门氏菌处理的 HT-29 细胞中的 ZO-1 和 p-IκB-α。总的来说,这项研究的结果表明,乳杆菌菌株通过有益地调节 TLR 负调节剂和 NF-κB 途径来减轻沙门氏菌诱导的炎症反应。