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目的运用基因敲除小鼠模型探讨脑源性神经营养因子(BDNF)在小鼠肠上皮屏障的作用。方法取BDNF+/+小鼠与BDNF+/-小鼠回肠及结肠黏膜,用透射电镜观察肠黏膜上皮的超微结构。免疫组织化学检测紧密连接蛋白ZO-1与occludin在结肠黏膜表达的变化,免疫印迹测定紧密连接蛋白claudin-1与claudin-2在结肠黏膜表达的差异。结果与BDNF+/+小鼠相比,BDNF+/-小鼠电镜下回肠上皮未见明显异常,结肠上皮屏障破坏,表现为微绒毛缺失、细胞间隙增宽以及细菌入侵;BDNF+/-小鼠结肠上皮细胞紧密连接蛋白occludin、ZO-1以及claudin-1表达下调,而claudin-2表达上调。结论 BDNF可能在小鼠结肠上皮屏障功能的发挥中起到调控作用。
Objective To investigate the effect of brain derived neurotrophic factor (BDNF) on the intestinal epithelial barrier in mice using a knockout mouse model. Methods The ileum and colonic mucosa of BDNF + / + mice and BDNF +/- mice were observed. The ultrastructure of intestinal mucosa was observed by transmission electron microscope. Immunohistochemistry was used to detect the expression of tight junction protein ZO-1 and occludin in colonic mucosa. Immunohistochemistry was used to detect the expression of claudin-1 and claudin-2 in colonic mucosa. Results Compared with BDNF + / + mice, there was no obvious abnormalities in the ileal epithelium of BDNF +/- mice under electron microscope. The barrier epithelium of the colon was damaged, which was characterized by lack of microvilli, widening of cell gap and bacterial invasion. BDNF +/- mice colonic epithelium The tight junction proteins occludin, ZO-1 and claudin-1 were down-regulated while claudin-2 was up-regulated. Conclusion BDNF may play a regulatory role in the development of mouse colonic epithelial barrier function.