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目的探讨肠上皮细胞O型糖链的合成阻滞对该细胞肠分化标记物MUC2表达及细菌黏附的影响。方法采用O型糖链抑制剂benzyl-α-GalNAc抑制结肠上皮细胞HT-29及其分化型细胞(HT-29-Gal)O型糖链的合成,经benzyl-α-GalNAc处理的HT-29和HT-29-Gal细胞分别命名为HT-29-OBN和HT-29-Gal-OBN。采用Real-time PCR和Western blotting方法检测上述4种细胞中MUC2基因的转录和蛋白表达水平,并将上述细胞与致病性大肠埃希菌(EPEC)和肠出血性大肠埃希菌(EHEC)O157:H7共培养,采用系列稀释菌落计数法观察细菌在上述细胞表面的黏附情况。结果 Realtime PCR和Western blotting结果显示,经benzyl-α-GalNAc处理后,HT-29和HT-29-Gal细胞MUC2的mRNA和蛋白表达均明显减少(P<0.05)。HT-29-OBN和HT-29-Gal-OBN细胞与致病性大肠埃希菌EPEC和EHEC O157:H7的黏附明显少于HT-29和HT-29-Gal细胞(P<0.05)。结论抑制肠上皮细胞O型糖链的合成可阻碍细菌黏附及MUC2的表达。
Objective To investigate the effects of O-glycosylation on the expression of MUC2 and bacterial adhesion in intestinal epithelial cells. Methods The O-type sugar chain inhibitor benzyl-α-GalNAc was used to inhibit the synthesis of HT-29 and HT-29-Gal O- And HT-29-Gal cells were named HT-29-OBN and HT-29-Gal-OBN, respectively. Real-time PCR and Western blotting were used to detect the transcription and protein expression of MUC2 in the above four cell lines. The above cells were incubated with pathogenic Escherichia coli (EECEC) and enterohemorrhagic Escherichia coli (EHEC) O157: H7 co-culture, the use of serial dilution colony count bacteria observed adhesion on the cell surface. Results Realtime PCR and Western blotting showed that mRNA and protein expression of MUC2 in HT-29 and HT-29-Gal cells were significantly decreased after benzyl-α-GalNAc treatment (P <0.05). The adhesion of HT-29-OBN and HT-29-Gal-OBN cells to pathogenic Escherichia coli EPEC and EHEC O157: H7 was significantly less than that of HT-29 and HT-29-Gal cells (P <0.05). Conclusion Inhibition of intestinal epithelial O-type sugar chain synthesis can hinder bacterial adhesion and MUC2 expression.