Bhatia Shikha, Prabhu P Nagendra, Benefiel Ann C, Miller Michael J, Chow JoMay, Davis Steven R, Gaskins H Rex
Department of Animal Sciences, University of Illinois at Urbana-Champaign, IL, USA.
Mol Nutr Food Res. 2015 Mar;59(3):566-73. doi: 10.1002/mnfr.201400639. Epub 2014 Dec 28.
Here we have tested the hypothesis that prebiotic galacto-oligosaccharides (GOS) may enhance mucosal barrier function through direct modulation of goblet cell function.
Human adenocarcinoma-derived LS174T cells, which exhibit an intestinal goblet cell-like phenotype, were used to examine the non-prebiotic effects of GOS on goblet cell functions. LS174T cells were treated with GOS, and the expression of goblet cell secretory product genes mucin 2 (MUC2), trefoil factor 3 (TFF3), resistin-like molecule beta (RETNLB) and the Golgi-sulfotransferase genes, carbohydrate (N-acetylglucosamine-6-O) sulfotransferase 5 (CHST5) and galactose-3-O-sulfotransferase 2 (GAL3ST2), was determined by real-time quantitative RT-PCR. In addition, the abundance of CHST5, TFF3 and RETNLB was confirmed by Western blot analysis. Following treatment with GOS for 72 h, the expression of MUC2 was significantly upregulated 2-4-fold, CHST5 and RETNLB, 5-7-fold, and TFF3 2-4-fold. Western blot analysis demonstrated increased abundance of RETNLB, TFF3 and CHST5. Addition of the Th2 cytokine IL-13 along with GOS resulted in synergistic induction of RETNLB and CHST5. IL-8 secretion was not affected by GOS treatment, suggesting that the effects of GOS are not mediated through an inflammatory pathway.
Collectively, the data indicate that GOS may enhance mucosal barrier function through direct stimulation of intestinal goblet cells.
在此我们验证了一个假设,即益生元低聚半乳糖(GOS)可能通过直接调节杯状细胞功能来增强黏膜屏障功能。
利用具有肠道杯状细胞样表型的人腺癌来源的LS174T细胞,来检测GOS对杯状细胞功能的非益生元效应。用GOS处理LS174T细胞,通过实时定量逆转录聚合酶链反应(RT-PCR)测定杯状细胞分泌产物基因粘蛋白2(MUC2)、三叶因子3(TFF3)、抵抗素样分子β(RETNLB)以及高尔基体硫酸转移酶基因碳水化合物(N-乙酰葡糖胺-6-O)硫酸转移酶5(CHST5)和半乳糖-3-O-硫酸转移酶2(GAL3ST2)的表达。此外,通过蛋白质印迹分析确认CHST5、TFF3和RETNLB的丰度。用GOS处理72小时后,MUC2的表达显著上调2至4倍,CHST5和RETNLB上调5至7倍,TFF3上调2至4倍。蛋白质印迹分析表明RETNLB、TFF3和CHST5的丰度增加。GOS与Th2细胞因子白细胞介素-13(IL-13)共同添加导致RETNLB和CHST5的协同诱导。IL-8分泌不受GOS处理的影响,这表明GOS的作用不是通过炎症途径介导的。
总体而言,数据表明GOS可能通过直接刺激肠道杯状细胞来增强黏膜屏障功能。