CAJ | 학술논문

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。
목적：본연구응용면역조직화학염색방법，관찰선천성거결장증（Hir －schsprunffs dis-ease，HD）환인유신경절세포장단、무신경절세포장단급비 HD 소인장벽（대조조）중수통도단백—1（AQP1）적표체결과，탐토 AQP—1재선천성거결장증적표체의의급기재 HD 발병궤리중가능기도적작용。방법수집강서성인동의원소인외과2009—2014년경수술절제적 HD 장조직표본65례，포괄확장단여경련단；장21례년령여지상방적비 HD 환인적수술절제결장표본작위대조조。분별대 HD 경련단、확장단、대조조장벽조직절편진행 HE 염색화 AQP1、조직단백매 D 화 S100적면역조직화학염색，통과계산궤성상계통조상존반，분별판정 AQP1재각조장조직점막하、기간신경총중양성세포염색면적적백분비，소득수거용 SPSS 19．0통계연건포진행처리분석。결과①대조조、HD 적확장단경 HE 화조직단백매 D 화 S100면역조화염색，균증실신경절세포존재；경련단미견신경절세포（도1，2）；②대조조、HD 경련단、확장단장조직중견 AQP1표체우소유혈관내피세포；③재정상결장、HD 확장단장조직점막하총、기간신경총중，신경절세포화설왕세포균가견 AQP1정양성표체，HD 경련단상응부위 AQP1표체대다정음성（90．2％），부소량정약양성표체；④반정량분석：AQP1재신경총중양성염색평분，HD경련단평분저우 HD 확장단평분，이자간차이구유통계학의의（P ＜0．01）。결론AQP1재 HD 경련단화정상결장、HD 확장단장벽신경총적표체유현저차이，아문시측재장도신경계통발육과정중，AQP1대신경절세포천이구유일정적조공작용，재 HD 적발병궤리중분연료일정각색병가능성위일충신적진단 HD 적신경표지물。
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 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.