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目的探索神经生长相关蛋白-43(GAP-43)在先天性巨结肠痉挛段和扩张段肠组织中的表达情况,进一步探索先天性巨结肠的发病机理。方法收集2012年1月至2013年6月期间因先天性巨结肠于深圳市儿童医院行巨结肠根治术的30例患儿的活体肠组织标本,行免疫组化染色及荧光定量聚合酶链反应(RT-PCR)以检测扩张段和狭窄段肠组织中GAP-43 m RNA及其蛋白的表达,并比较扩张段和狭窄段肠组织中两者表达的差异。结果RT-PCR结果显示,痉挛组肠组织中GAP-43 m RNA的表达水平的中位数为0.052 8,低于扩张组的0.119 0(P<0.05)。免疫组化染色结果显示:30例患儿的狭窄段肠组织和扩张段肠组织的肌间神经丛及黏膜下神经丛中GAP-43蛋白均呈阳性表达,但痉挛段黏膜下神经丛和肌间神经丛的染色均较扩张段相应部位浅;与扩张组的黏膜下神经丛和肌间神经丛比较,痉挛组相应部位的GAP-43蛋白的平均光密度值均较低(P<0.05)。结论先天性巨结肠狭窄段肠组织中GAP-43蛋白的表达较扩张段肠组织下调,提示GAP-43蛋白可能是先天性巨结肠发病的危险因素之一。
Objective To explore the expression of GAP-43 in Hirschsprung’s disease and dilatation of intestinal tract and to explore the pathogenesis of Hirschsprung’s disease. Methods Thirty cases of children with Hirschsprung’s disease treated with Hirschsprung’s disease at Shenzhen Children’s Hospital from January 2012 to June 2013 were collected for immunohistochemical staining and fluorescent quantitative polymerase chain reaction (RT-PCR) was used to detect the expression of GAP-43mRNA and its protein in the expanded and narrow intestine tissues, and the difference between the expression of GAP-43mRNA and its expression in the expanded and narrow intestine tissues was compared. Results The RT-PCR results showed that the median of GAP-43 mRNA expression in spastic group was 0.052 8, which was lower than that of the expansion group (0.119 0, P <0.05). The results of immunohistochemistry showed that the expression of GAP-43 protein in the myenteric plexus and submucosal plexus of the stenosis intestinal segment and dilated intestinal segment of 30 cases was positive, but the submucosal plexus and muscle Compared with the submucosal plexus and myenteric plexus in the dilation group, the mean optical density of GAP-43 protein in the corresponding part of the spastic group was lower (P <0.05) . Conclusions The expression of GAP-43 protein in intestinal tissue of Hirschsprung’s disease is down-regulated compared with that of the expanded gut, suggesting that GAP-43 may be one of the risk factors for the pathogenesis of Hirschsprung’s disease.