不同母乳低聚糖对肠道LS174T杯状细胞黏蛋白分泌相关基因表达的影响

Effects of different human milk oligosaccharides on the expression of mucin secretion-related genes in intestinal LS174T goblet cells

中文关键词:  母乳低聚糖  LS174T杯状细胞  2’-岩藻糖基乳糖  3’-唾液酸乳糖  低聚半乳糖  黏蛋白2  黏蛋白分泌相关基因

英文关键词:human milk oligosaccharides  LS174T goblet cells  2’-fucosyllactose  3’-sialyllactose  galactooligosaccharide  mucin 2  mucin secretion-related genes

中文摘要:

      目的 分析不同母乳低聚糖对肠道LS174T杯状细胞黏蛋白分泌相关基因表达的影响。方法 以人肠道LS174T细胞为研究对象, 将2’-岩藻糖基乳糖(2’-fucosyllactose, 2’-FL)、3’-唾液酸乳糖(3’-sialyllactose, 3’-SL)和低聚半乳糖(galactooligosaccharide, GOS) 3种低聚糖和乳糖对细胞的作用时间分成24 h组和48 h组, 分别在细胞稳态和细胞炎症状态下, 以黏蛋白2(mucin 2, MUC2)及其相关基因[三叶因子-3 (trefoil factor 3, TFF3)、抵抗素样蛋白B (recombinant resistinlike beta, RETNLB)、碳水化合物磺基转移酶5 (carbohydrate sulfotransferase 5, CHST5)、半乳糖-3-O-磺基转移酶2 (galactose-3-O-sulfotransferase 2, GAL3ST2)]的表达水平为评价指标, 通过实时荧光定量多聚核苷酸链式反应(real time-quantitative polymerase chain reaction, RT-qPCR)试验确定影响肠道LS174T杯状细胞黏蛋白分泌相关基因表达的母乳低聚糖。结果 在正常状态下, GOS作用后, MUC2及黏蛋白分泌相关基因表达量无显著变化(P>0.05); 作用48 h时, 3’-SL可显著提高RETNLB基因表达量(P<0.05); 在炎症状态下, 2’-FL和GOS可极显著促进MUC2、CHST5和TFF3的表达(P<0.01), 表明在炎症状态下, 2’-FL和GOS可促进杯状细胞分泌黏蛋白。结论 在细胞炎症状态下, 2’-FL和GOS可提高肠道LS174T杯状细胞黏蛋白分泌相关基因的表达量。

英文摘要:

      Objective To analyze the effects of different human milk oligosaccharides on the expression of mucin secretion-related genes in intestinal LS174T goblet cells. Methods Taking LS174T cells of human intestine as the research object, the effect times of 3 kinds of oligosaccharides and lactose [(2’-fucosyllactose, 2’-FL), (3’-sialyllactose, 3’-SL) and (galactooligosaccharide, GOS)] on cells were divided into 24 h and 48 h group. The expression levels of mucin 2 (MUC2) and its related genes [trefoil factor 3 (TFF3), recombinant resistinlike beta (RETNLB), carbohydrate sulfotransferase 5 (CHST5), galactose-3-O-sulfotransferase 2 (GAL3ST2)] were taken as evaluation indicators under the conditions of cell homeostasis and cell inflammation, respectively. The human milk oligosaccharides that affect the expression of mucin secretion related genes of LS174T goblet cells in the intestine were determined by real time-quantitative polymerase chain reaction (RT-qPCR). Results In homeostasis, the expressions of MUC2 and mucin secretion-related genes were not significantly changed after GOS treatment (P>0.05). After treatment for 48 h, 3’-SL significantly increased the expression of RETNLB gene (P<0.05). In inflammatory state, 2’-FL and GOS significantly promoted the expression of MUC2, CHST5 and TFF3 (P<0.01), indicating that 2’-FL and GOS could promote mucin secretion of goblet cells in inflammatory state. Conclusion In inflammatory state, 2’-FL and GOS can increase the expression of mucin secretion related genes in intestinal LS174T goblet cells.