Immunoendocrinology, Division of Medical Biology, Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Hanzeplein 1, RB, Groningen, 9700, The Netherlands.
Drug Design XB20, Groningen Research Institute of Pharmacy, University of Groningen, AD, Groningen, 9700, The Netherlands.
Mol Nutr Food Res. 2021 Apr;65(7):e2000425. doi: 10.1002/mnfr.202000425. Epub 2021 Mar 3.
Human milk oligosaccharides (hMOs) can attenuate inflammation by modulating intestinal epithelial cells, but the mechanisms of action are not well-understood. Here, the effects of hMOs on tumor necrosis factor-α (TNF-α) induced inflammatory events in gut epithelial cells are studied.
The modulatory effects of 2'-fucosyllactose, 3-fucosyllactose (3-FL), 6'-sialyllactose, lacto-N-tetraose, lacto-N-neotetraose (LNnT), lactodifucotetraose (LDFT), and lacto-N-triaose (LNT2) on immature (FHs 74 Int) and adult (T84) intestinal epithelial cells with or without TNF-α are determined. Interleukin-8 (IL-8) secretion in FHs 74 Int and T84 are quantified to determine hMO induced attenuation of inflammatory events by ELISA. 3-FL, LNnT, and LDFT significantly attenuate TNF-α induced inflammation in FHs 74 Int, while LNT2 induces IL-8 secretion in T84. In addition, microscale thermophoresis assays and ELISA are used to study the possible mechanisms of interaction between effective hMOs and tumor necrosis factor receptor 1 (TNFR1). 3-FL, LNnT, and LDFT exert TNFR1 ectodomain shedding while LNnT also shows binding affinity to TNFR1 with a Kd of 900 ± 660 nM.
The findings indicate that specific hMO types attenuate TNF-α induced inflammation in fetal gut epithelial cells through TNFR1 in a hMO structure-dependent fashion suggest possibilities to apply hMOs in management of TNF-α dependent diseases.
人乳寡糖（hMOs）可以通过调节肠道上皮细胞来减轻炎症，但作用机制尚不清楚。本研究旨在研究 hMOs 对肠道上皮细胞中肿瘤坏死因子-α（TNF-α）诱导的炎症事件的影响。
研究了 2'-岩藻糖基乳糖、3-岩藻糖基乳糖（3-FL）、6'-唾液酸乳糖、乳-N-四糖、乳-N-新四糖（LNnT）、乳二岩藻四糖（LDFT）和乳-N-三糖（LNT2）在有无 TNF-α 的情况下对未成熟（FHs 74 Int）和成人（T84）肠道上皮细胞的调节作用。通过 ELISA 定量测定 FHs 74 Int 和 T84 中的白细胞介素-8（IL-8）分泌，以确定 hMO 对炎症事件的抑制作用。3-FL、LNnT 和 LDFT 显著减轻了 FHs 74 Int 中 TNF-α 诱导的炎症，而 LNT2 则诱导了 T84 中 IL-8 的分泌。此外，使用微量热泳动分析和 ELISA 研究了有效 hMO 与肿瘤坏死因子受体 1（TNFR1）之间相互作用的可能机制。3-FL、LNnT 和 LDFT 发挥了 TNFR1 外显肽脱落的作用，而 LNnT 也显示出与 TNFR1 的结合亲和力，Kd 值为 900±660 nM。
研究结果表明，特定的 hMO 类型通过 TNFR1 以 hMO 结构依赖性的方式减轻胎儿肠道上皮细胞中 TNF-α 诱导的炎症，这为应用 hMO 治疗 TNF-α 依赖性疾病提供了可能性。
