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叶钧,宋丽丽,刘韵,田音,潘琼,彭志红,汪荣泉.肠分化细胞黏蛋白O-型糖链合成抑制导致MUC2表达减低以及对细菌侵袭易感[J].陆军军医大学学报(原第三军医大学学报),2013,35(19):2056-2059.
Ye Jun,Song Lili,Liu Yun,Tian Yin,Pan Qiong,Peng Zhihong,Wang Rongquan.Inhibition of O-glycan chains synthesis in intestinal differentiated epithelial cells induces lower MUC2 expression and susceptibility to bacterial invasion[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(19):2056-2059.
叶钧,宋丽丽,刘韵,田音,潘琼,彭志红,汪荣泉.肠分化细胞黏蛋白O-型糖链合成抑制导致MUC2表达减低以及对细菌侵袭易感[J].陆军军医大学学报(原第三军医大学学报),2013,35(19):2056-2059. DOI:
Ye Jun,Song Lili,Liu Yun,Tian Yin,Pan Qiong,Peng Zhihong,Wang Rongquan.Inhibition of O-glycan chains synthesis in intestinal differentiated epithelial cells induces lower MUC2 expression and susceptibility to bacterial invasion[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(19):2056-2059. DOI:
目的 探讨O-型糖链合成的抑制对肠分化细胞内细菌侵袭数量和细胞内MUC2表达水平的影响。 方法 对肠分化细胞(HT-29-Gal)用O-型糖链抑制剂(benzyl-N-acetyl-α-D-galactosaminide,benzyl-α-GalNAc)处理,采用Real-time PCR和Western blot检测MUC2 mRNA和蛋白的表达情况。并将HT-29-Gal细胞及benzyl-α-GalNAc 处理的HT-29-Gal细胞分别与肠致病性大肠杆菌(enteropathogenic E.coli,EPEC)和肠出血性大肠杆菌(EHEC O157∶H7) 37 ℃孵育2 h, 再加入100 μg/mL的庆大霉素,杀灭细胞外及粘附于细胞表面的细菌。最后采用系列稀释克隆计数法观察benzyl-α-GalNAc 处理的HT-29-Gal细胞对细菌侵袭的影响。 结果 Real-time PCR和Western blot检测发现经benzyl-α-GalNAc处理的HT-29-Gal细胞MUC2的mRNA和蛋白表达水平明显降低(
P
0.05)。侵袭入benzyl-α-GalNAc 处理的HT-29-Gal细胞的EPEC和EHEC O157∶H7的数量较对照细胞显著增加(
P
0.05)。 结论 抑制HT-29-Gal细胞黏蛋白O-型糖链的合成导致侵袭入细胞内细菌数量增加和MUC2的表达降低。
Objective To determine the effect of inhibiting O-glycan chains synthesis in intestinal differentiated epithelial cells on the bacteria invasion into the cells and MUC2 expression. Methods The intestinal differentiated cell line HT-29-Gal was treated with O-glycan chains synthesis inhibitor
benzyl-N-acetyl-α-D-galactosaminide (benzyl-α-GalNAc). The ex
pression of MUC2 at mRNA and protein levels was detected by real-time PCR and Western blotting. The HT-29-Gal cells treated with or without benzyl-α-GalNAc were co-cultured with enteropathogenic E.coli (EPEC)and enterohemorrhage E.coli (EHEC O157∶H7)for 2 h
then the gentamicin (100 μg/mL) was added in the culture medium for 2 h to exclude extracellular and cellular surface-adherent bacteria after 3 times of washing with PBS. Finally
serial dilution cloning counting was used to observe the numbers of invaded bacteria into the HT-29-Gal cells treated with or without benzyl-α-GalNAc. Results Real-time PCR and Western blotting indicated that the mRNA and protein Levels of MUC2 were significantly lower in HT-29-Gal cells treated with the benzyl-α-GalNAc than the untreated HT-29-Gal cells(
P
0.05).The numbers of EPEC and EHEC O157∶H7 invaded into the HT-29-Gal cells exposed to benzyl-α-GalNAc were significantly larger than those untreated (
P
0.05 ). Conclusion Inhibition of O-glycan chains synthesis in HT-29-Gal cells leads more bacteria to invade into the cells and lower expression of MUC2 at both mRNA and protein levels.
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