免疫组织化学染色在诊断先天性巨结肠和类缘病中的应用

The application of immunohistochemistry in the diagnosis of Hirschsprung＇s disease and allied diseases

目的探讨免疫组织化学染色方法在先天性巨结肠和类缘病中的应用。方法收集上海儿童医学中心近年来的6例先天性巨结肠（HSCD）、2例肠神经元性发育异常症（IND）、14例HSCD＋IND和4例神经节发育不良症的手术切除标本，4例对照标本和5例肠黏膜活检标本。选择蛋白基因产物9．5（protein gene product9．5，PGP9．5）、组织蛋白酶D（cathepsinD）和钙视网膜蛋白（calretinin，CR）作为单抗，免疫组化染色，观察黏膜、黏膜下层和肌层的染色情况。结果正常肠壁黏膜下层和肌间神经节细胞、神经纤维束对PGP9．5均显著着色，黏膜层，黏膜下层可见较多纤细的神经纤维。组织蛋白酶D仅对神经节细胞染色，成熟的神经节细胞染色强，未成熟的神经节细胞染色弱；神经纤维不表达。CR神经节细胞有表达，正常肠壁黏膜层和黏膜下层可见中等量纤细的神经纤维。HSCD、IND和神经节发育不良症肠壁的PGP9．5、组织蛋白酶D和CR都有各自的不同表达。结论本组研究显示CR免疫组化可用于直肠黏膜吸引活检病理诊断先天性巨结肠；PGP9．5和CR在辨认未成熟神经节细胞方面优于组织蛋白酶D染色；PGP9．5、组织蛋白酶D和CR免疫组化染色有辅助病理诊断HSCD、IND和肠神经节发育不良症的作用。

Objective To study the use of immunohistochemistry in the diagnosis of Hirschsprung＇s disease （HSCD） and allied diseases. Methods Specimens were collected from resected colons of 6 HSCD, 2 intestinal neuronal dysplasia （IND）, 14 HSCD ＋ IND, 4 hypoganglionosis patients,4 control cases and 5 cases of mucosal biopsies, from Shanghai Children＇s Medical Center. The expressions of protein gene product 9. 5 （PGP 9. 5）, cathepsin D and calretinin （CR） were measured using immunohistochimostry （IHC）. Results Ganglion cells and nerve fibers of submucosa and muscle layers were positively stained for PGP 9. 5 in normal intestines. A lot of positive staining of fine nerve fibers was seen. Only ganglion cells were positively stained for cathepsin D. Staining of mature ganglion cells were much stronger than the immature cells. The nerve fibers showed no expression. The ganglion cells and the nearby nerve fibers were positively stained for CR, with moderate fine nerve fibers in the normal mucosa and submucosa. There were differential expressions of PGP 9. 5, cathepsin D and CR among HSCD, IND and hypoganglionosis patients. Conclusions This study shows that CR IHC may be a useful technique in the pathological diagnosis of suction rectal biopsies for HSCD. PGP 9. 5 and CR IHC stain are better than cathepsin D in recognizing immature ganglion cells. PGP 9. 5, cathepsin D and CR IHC stain are useful adjunct techniques for HSCD, IND and hypoganglionsis.