先天性巨结肠患儿术后肠道组织中CAD、SOX2的表达水平及其临床意义

Expression levels and clinical significance of cathepsin D and sex-determining region Y-box protein 2 in intestinal tissues of children with Hirschsprung’s disease after operation

目的探讨先天性巨结肠(HD)患儿术后肠道组织蛋白酶D(CAD)、性别决定区Y框蛋白2(SOX2)的表达水平及其临床意义。方法选取2015年5月至2019年3月郑州大学第一附属医院收治的56例HD患儿结肠组织(HD组)和23例因肠道梗阻或穿孔实施造瘘手术获取的结肠组织标本(对照组)。采用免疫组化、Western-blot技术检测结肠组织中CAD、SOX2的表达水平,并采用Pearson线性相关分析法分析CAD、SOX2表达与病变肠段肌间神经丛直径、神经节细胞数的关系。结果HD患儿的狭窄段组织中CAD蛋白、SOX2蛋白阳性表达率分别为7.14%、12.50%,移行段肠组织中CAD蛋白、SOX2蛋白阳性表达率分别为32.14%、35.71%,明显低于对照组的78.26%、82.61%,而狭窄段组织中CAD蛋白、SOX2蛋白阳性表达率明显低于移行段,差异均有统计学意义(P<0.05);HD患儿的狭窄段肠组织中肌间神经丛直径、神经节细胞数目分别为(30.66±5.14)μm、(0.83±0.24)个/视野,移行段肠组织中肌间神经丛直径、神经节细胞数目分别为(31.20±5.83)μm、(1.94±0.56)个/视野,明显短(少)于对照组的(54.29±8.50)μm、(5.40±1.84)个/视野,狭窄段肠组织中肌间神经丛直径、神经节细胞数目明显短(少)于移行段,差异均有统计学意义(P<0.05);Pearson相关分析结果显示,HD患儿狭窄段、移行段肠组织中肌间神经丛直径、神经节细胞数目与CAD蛋白、SOX2蛋白表达强度均呈显著正相关(P<0.05)。结论HD患儿病变结肠组织中CAD蛋白、SOX2蛋白的表达强度下调,可能与肌间神经丛直径、神经节细胞数目的减少有关。

Objective To investigate the expression levels and clinical significance of cathepsin D(CAD)and sex-determining region Y-box protein 2(SOX2)in intestinal tissues of children with Hirschsprung’s disease(HD)after operation.Methods Colon tissue samples from 56 children of colon tissue of HD children(HD group)and 23 colon tissue samples obtained by colostomy due to intestinal obstruction or perforation(control group)in the First Affiliated Hospital of Zhengzhou University from May 2015 to March 2019 were selected.Immunohistochemistry and Western-blot technique were used to detect the expression levels of CAD and SOX2 in colon tissue.Pearson linear correlation analysis was used to analyze the relationship between the expression of CAD and SOX2 and the diameter of intermuscular nerve plexus and the number of ganglion cells in the diseased intestinal segment.Results The positive expression rates of CAD protein and SOX2 protein in the narrow segment of HD children were 7.14%and 12.50%,respectively,and those in the transitional segment of intestinal tissue were 32.14%and 35.71%,respectively,which were significantly lower than 78.26%and 82.61%in the control group;the two indexes in the narrow segment were significantly lower than those in the transitional segment;the differences were statistically significant(P<0.05).The diameter of intermuscular nerve plexus and the number of ganglion cells in the narrow segment of intestinal tissue of HD children were(30.66±5.14)μm,(0.83±0.24)/field of view,and those in the transitional segment of intestinal tissue were(31.20±5.83)μm and(1.94±0.56)/field of view,which were significantly shorter(less)than(54.29±8.50)μm,(5.40±1.84)/field of view in the control group;the two indexes in the narrow segment of intestinal tissue were significantly shorter(less)than those in the transitional segment;the differences were statistically significant(P<0.05).Pearson correlation analysis showed that the diameter of myenteric plexus and the number of ganglion cells in the intestinal tissue of the transitional segment were positively correlated with the expression of CAD protein and SOX2 protein(P<0.05).Conclusion The expression of CAD protein and SOX2 protein in the colon tissue of children with HD is down-regulated,which may be related to the decrease of the diameter of the myenteric plexus and the number of ganglion cells.