Division of Nutritional Sciences, University of Illinois, 1207 W. Gregory Dr. Urbana, Urbana, IL 61801, USA.
Exp Biol Med (Maywood). 2011 Dec;236(12):1402-12. doi: 10.1258/ebm.2011.011186. Epub 2011 Nov 18.
The signals that mediate goblet cell expression of specific mucin chemotypes are poorly defined. Animal and in vitro studies show that acidomucin chemotypes may be altered by inflammation and changes in intestinal microbiota. To examine factors that may elicit this response, human adenocarcinoma-derived LS174T cells, which have a goblet cell-like phenotype and produce both sulfo- and sialomucins, were used to examine the effects of selected microbial and host factors on expression of goblet cell secretory product genes, sulfotransferases and sulfomucin production. Expression of genes encoding mucin 2 (MUC2), resistin-like molecule β (RETNLB), and trefoil factor 3 (TFF3) and Golgi sulfotransferases, carbohydrate (N-acetylglucosamine 6-O) sulfotransferase 5 (CHST5) and galactose-3-O-sulfotransferase 2 (GAL3ST2), was measured by quantitative reverse transcriptase-polymerase chain reaction following treatment with bacterial flagellin, tumor necrosis factor α (TNF-α) or the mucogenic cytokine interleukin-13 (IL-13). Expression of the toll-like receptor 5 (TLR5) gene was also analysed. Sulfomucin expression was examined via high-iron diamide/alcian blue (HID/AB) histochemistry and immunofluorescent staining for the Sulfo Le(a) antigen, which is synthesized in part by GAL3ST2. Flagellin, IL-13 and TNF-α all significantly increased GAL3ST2, MUC2, TFF3 and TLR5 expression, while only IL-13 increased RETNLB and CHST5 expression. Based on HID/AB histochemistry, mucin sulfation was significantly increased in response to both flagellin and IL-13 but not TNF-α. Only treatment with flagellin increased the expression of the Sulfo Le(a) antigen. Collectively, these results indicate that bacterial flagellin, IL-13 and TNF-α differentially modulate the expression of goblet cell secretory product genes, sulfotransferases and sulfomucin production.
介导杯状细胞表达特定粘蛋白化学型的信号通路尚未完全明确。动物和体外研究表明，粘蛋白酸化学型可能会受到炎症和肠道微生物群变化的影响而发生改变。为了研究可能引发这种反应的因素，我们使用人结肠癌细胞系 LS174T 细胞（具有杯状细胞样表型，可同时产生硫酸乙酰肝素和唾液酸粘蛋白），研究了选定的微生物和宿主因素对杯状细胞分泌产物基因、硫酸转移酶和硫酸粘蛋白产生的影响。采用定量逆转录聚合酶链反应（qRT-PCR）检测基因 MUC2（粘蛋白 2）、RETNLB（抵抗素样分子β）和 TFF3（三叶因子 3）、高尔基硫酸转移酶 CHST5（碳水化合物（N-乙酰氨基葡萄糖 6-O）硫酸转移酶 5）和 GAL3ST2（半乳糖-3-O-硫酸转移酶 2）的表达，细胞先用细菌鞭毛蛋白、肿瘤坏死因子-α（TNF-α）或粘蛋白生成细胞因子白细胞介素-13（IL-13）处理后进行测量。还分析了 TLR5（Toll 样受体 5）基因的表达。通过高铁二胺/阿辛蓝（HID/AB）组织化学和 Sulfo Le(a)抗原的免疫荧光染色来检测硫酸粘蛋白的表达，Sulfo Le(a)抗原部分由 GAL3ST2 合成。鞭毛蛋白、IL-13 和 TNF-α 均显著增加了 GAL3ST2、MUC2、TFF3 和 TLR5 的表达，而只有 IL-13 增加了 RETNLB 和 CHST5 的表达。根据 HID/AB 组织化学，鞭毛蛋白和 IL-13 均显著增加了粘蛋白的硫酸化，但 TNF-α 没有。只有用鞭毛蛋白处理才能增加 Sulfo Le(a)抗原的表达。综上所述，这些结果表明，细菌鞭毛蛋白、IL-13 和 TNF-α 可差异化地调节杯状细胞分泌产物基因、硫酸转移酶和硫酸粘蛋白的产生。
