AQP1在先天性巨结肠肠壁的表达及其意义

The expression and its significance of AQP-1 in the intestinal wall of Hirschsprung′s disease

目的：本研究应用免疫组织化学染色方法，观察先天性巨结肠症（Hir －schsprunffs dis-ease，HD）患儿有神经节细胞肠段、无神经节细胞肠段及非 HD 小儿肠壁（对照组）中水通道蛋白—1（AQP1）的表达结果，探讨 AQP—1在先天性巨结肠症的表达意义及其在 HD 发病机理中可能起到的作用。方法收集江西省儿童医院小儿外科2009—2014年经手术切除的 HD 肠组织标本65例，包括扩张段与痉挛段；将21例年龄与之相仿的非 HD 患儿的手术切除结肠标本作为对照组。分别对 HD 痉挛段、扩张段、对照组肠壁组织切片进行 HE 染色和 AQP1、组织蛋白酶 D 和 S100的免疫组织化学染色，通过计算机成像系统照相存盘，分别判定 AQP1在各组肠组织黏膜下、肌间神经丛中阳性细胞染色面积的百分比，所得数据用 SPSS 19．0统计软件包进行处理分析。结果①对照组、HD 的扩张段经 HE 和组织蛋白酶 D 和 S100免疫组化染色，均证实神经节细胞存在；痉挛段未见神经节细胞（图1，2）；②对照组、HD 痉挛段、扩张段肠组织中见 AQP1表达于所有血管内皮细胞；③在正常结肠、HD 扩张段肠组织黏膜下丛、肌间神经丛中，神经节细胞和雪旺细胞均可见 AQP1呈阳性表达，HD 痉挛段相应部位 AQP1表达大多呈阴性（90．2％），仅少量呈弱阳性表达；④半定量分析：AQP1在神经丛中阳性染色评分，HD痉挛段评分低于 HD 扩张段评分，二者间差异具有统计学意义（P ＜0．01）。结论AQP1在 HD 痉挛段和正常结肠、HD 扩张段肠壁神经丛的表达有显著差异，我们猜测在肠道神经系统发育过程中，AQP1对神经节细胞迁移具有一定的调控作用，在 HD 的发病机理中扮演了一定角色并可能成为一种新的诊断 HD 的神经标志物。

Objective To explain the possible pathogenesis of ENS defection in HD,we now reveal the abnormal expression of a water channel protein,which named Aquaporin—1（AQP—1）,in intestinal wall of Hir-schsprung＇s disease and the potential role AQP—1 played in the development of HD.Methods Specimens were collected from resected colon in both aganglionic bowel segments and ganglionic segments of 65 HD patients. The Colon in the control group colon was resected from 21 age-matched patients without HD.All specimens un-derwent haematoxylin and eosin（HE）staining,followed by immunostaining for AQP—1、Cathepsin D and S—100. Positively stained area density was evaluated at 400 magnification,calculating the sum of the areas which occu-pied on the basis of the positively stained area in plexuses.Results ①The ganglionic cells were detected in the dilated Segments of HD and tissue of control group,which is failed in narrow segments by HE,also in immu-nohistochemical staining of Cathepsin-D and S — 100;②AQP — 1 was strongly positive in Endothelial cells both within the aganglionic and ganglionic gut and control group.③In the ganglionic cells and Schwann Cells of the dilated segments,AQP — 1 was positive both in the Auerbach plexuses and Meissner plexuses but negative in most of（about 90.2%）the narrow group.④Semi-quantitative analysis showed that the positive staining area＆amp;nbsp;percentage of AQP— 1 has significant differences between the narrow segments and the dilated segments of HD （P 〈0.001）. Conclusions AQP — 1 immunohistochemical staining was strongly positive in ganglionic cells both in the dilated segments group and the normal colon but mostly negative in the narrow segments group.We presumably that AQP—1 may play a important role in the intestinal nerve cells migration to induce the develop-ment of HD thus AQP— 1can be useful as one of the neuronal markers for the diagnosis of HD.