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目的研究先天性巨结肠症(HD)患儿各段肠管中Neuroligin2基因的表达情况,探讨Neuroligin2表达与HD发生的可能关系。方法选取山东大学齐鲁医院小儿外科行手术治疗的HD患儿结肠标本共42例,将HD患儿正常段设为正常对照组,移行段及狭窄段设为实验组,应用Western blotting、实时荧光定量聚合酶链式反应(qRT-PCR)及免疫组织化学染色方法观察Neuroligin2在各肠段的表达情况。结果 Western blotting结果显示Neuroligin2蛋白在狭窄段相对表达量为0.012±0.001,明显少于移行段和正常段(0.039±0.001,0.057±0.002;P<0.05)。qRT-PCR结果显示,Neuroligin2的mRNA水平与蛋白表达趋势一致。免疫组织化学染色示HD患儿正常段肠管的肌间肠神经节细胞中有Neuroligin2强阳性细胞的表达,而在狭窄段肠管中弱阳性表达,移行段肠管可见Neuroligin2阳性细胞的表达。结论 HD患儿狭窄段肠管中Neuroligin2的表达缺失可能与HD的发生有关。
Objective To investigate the expression of Neuroligin2 in various stages of intestine in children with Hirschsprung ’s disease (HD) and to explore the possible relationship between Neuroligin2 expression and HD. Methods A total of 42 colon specimens of children with HD underwent surgery in pediatric surgery department of Qilu Hospital of Shandong University. The normal control group, the transitional section and the stenotic segment of HD group were selected as experimental group. Western blotting, real-time fluorescence quantitative Polymerase chain reaction (qRT-PCR) and immunohistochemical staining were used to observe the expression of Neuroligin2 in various segments of the bowel. Results The results of Western blotting showed that the relative expression of Neuroligin2 protein in the stenotic segment was 0.012 ± 0.001, which was significantly lower than that in the normal and normal segments (0.039 ± 0.001, 0.057 ± 0.002; P <0.05). qRT-PCR results showed that the mRNA level of Neuroligin2 was consistent with the trend of protein expression. Immunohistochemical staining showed that Neuroligin2 strongly positive cells were expressed in the intestine ganglion cells in the normal intestine of children with HD, but weakly positive in the narrow intestine. The expression of Neuroligin2 positive cells in the bowel of migrating segment was observed. Conclusion The loss of expression of Neuroligin2 in the narrow intestine of children with HD may be related to the occurrence of HD.