Bifidobacterium longum subsp. longum K5 alleviates inflammatory response and prevents intestinal barrier injury induced by LPS in vitro based on comparative genomics
2022
Li Zhao | Qinggang Xie | Smith Etareri Evivie | Yingxue Yue | Han Yang | Xiuli Lv | Fei Liu | Bailiang Li | Guicheng Huo
Excessive immune responses and intestinal barrier injury are leading causes of inflammatory bowel diseases (IBD). Previously, we demonstrated that B. longum subsp. longum K5 alleviated DSS-stimulated colitis in mice, while K15 could not. Here, we investigated both strains' underlying mechanisms regarding their anti-inflammatory and intestinal barrier protective capacities. We assessed the potential characters of two strains, anti-inflammation activities in the LPS-induced RAW 264.7 cell model, and intestinal epithelial barrier functions in the LPS-induced Caco-2 cell monolayers model. Results showed that B. longum subsp. longum K5 exhibited high inhibition against the adhesion of E. coli ATCC 25922 to HT-29 and acetic acid production. B. longum subsp. longum K5 (MOI 1:100) significantly decreased the pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) and inflammatory mediator ROS, NO level in LPS (1 μg/mL) -induced RAW 264.7 cells. It also significantly increased TEER value and decreased paracellular permeability of Caco-2 cells stimulated by LPS. B. longum subsp. longum K5 (MOI 1:100) up-regulated ZO-1, occlaudin and claudin-1 mRNA expression, down-regulated the mRNA expression level of TLR4 and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) in LPS (100 μg/mL) -stimulated Caco-2 cells. While B. longum subsp. longum K15 did not do as well as B. longum subsp. longum K5 on these assays. Furthermore, the whole-genome sequencing results revealed a gene encoding fatty acid synthase associated with the immune system in K5 specific genes, LacI family related with carbohydrate metabolism, and serine/threonine-protein kinase involved in signal transduction with an amino acid mutation. Our findings provided the theoretical basis for B. longum subsp. longum K5 to be used as a candidate for IBD therapy.
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