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目的研究肠道菌群失衡对小鼠肠黏膜机械屏障的影响,探讨优势菌群与肠道黏膜屏障的相互作用机制。方法利用ELISA法检测肠黏膜sIgA含量变化,RT-PCR技术及免疫组化法检测肠黏膜上皮细胞Mucin2、Mucin3和防御素mRNA转录产物的变化。结果菌群失衡小鼠小肠段肠黏膜sIgA水平下降,重度菌群失衡组与正常组比较下降显著(P<0.05),与轻度菌群失衡组相比下降明显。菌群失衡小鼠上皮细胞中的Mucin2、Mucin3和防御素mRNA转录产物与正常小鼠相比明显增高,在杯状细胞胞浆内Mucin2蛋白阳性表达增强,且重度菌群失衡组与轻度菌群失衡组小鼠比较,Mucin2、Mucin3和防御素增高明显。结论菌群失衡可导致肠黏膜sIgA分泌量随菌群失衡程度呈下降趋势,同时引起黏液层中黏液素和防御素分泌量增加。
Objective To study the effect of intestinal microflora imbalance on the mechanical barrier of intestinal mucosa in mice and to explore the interaction mechanism between dominant bacteria and intestinal mucosal barrier. Methods The changes of sIgA in intestinal mucosa were detected by ELISA. The mRNA expressions of Mucin2, Mucin3 and defensin in intestinal epithelial cells were detected by RT-PCR and immunohistochemistry. Results The intestinal mucosa sIgA level in the small intestine of mice with imbalanced flora decreased significantly, while that in severe imbalance group decreased significantly compared with the normal group (P <0.05), and decreased significantly compared with the mild imbalance group. Mucin2, Mucin3 and defensin mRNA transcripts in the imbalance mice group were significantly higher than those in normal mice, and Mucin2 protein expression was positive in the goblet cell cytoplasm. Mucin2, Mucin3 and defensin significantly increased in the imbalance group mice. Conclusion The imbalance of flora may lead to the decrease of sIgA secretion in intestinal mucosa with the degree of imbalance in the flora, and increase the secretion of mucin and defensin in the mucous layer.