*Pediatric Surgery, Department of Women's and Children's Health, University of Padova †General Pathology and Cytopathology, Section of Medical-Diagnostic Sciences and Special Therapies, Department of Medicine, University of Padova, Padova, Italy.
J Pediatr Gastroenterol Nutr. 2013 Dec;57(6):780-7. doi: 10.1097/MPG.0b013e3182a934c7.
The aim of this study was to investigate calretinin and β-tubulin immunohistochemical expression together with submucosal nerve trunks morphology in differently innervated segments of Hirschsprung disease (HD) and total colonic aganglionosis (TCA).
A total of 25 cases (22 HD, 3 TCA) and 18 controls were processed for calretinin and β-tubulin immunohistochemistry. Sections representative of distal aganglionic, transition, and proximal ganglionic segments were evaluated by a visual grading score; β-tubulin was evaluated also by image analysis. Submucosal nerve trunks hypertrophy and hyperplasia were measured by citomorphology. The length of proximal segment was correlated to postoperative bowel function.
Controls showed intense calretinin and β-tubulin staining. In HD and TCA, calretinin staining was related to the presence of ganglion cells: negative in distal, faint in transition, intense in proximal segment. β-Tubulin staining was weak in all of the segments of HD and negative in TCA. Hypertrophic and hyperplastic nerve trunks characterized aganglionic segment, and progressively decreasing nerve size was observed in transition and ganglionic segments. Transient postoperative constipation, soiling, or enterocolitis was present in 59% of patients with HD without clear relation to proximal segment length or presence of hypertrophic nerve trunks.
Calretinin is a reliable marker of the presence of ganglion cells, and, together with nerve hypertrophy, it helps to identify the transition zone. Length and nerve size of proximal segment in resected specimen did not affect the postsurgical intestinal function. Reduced β-tubulin expression along the entire colonic tract, included proximal ganglionic segments, may represent a potential impairing factor for the enteric neural transmission.
本研究旨在探讨钙视网膜蛋白和β-微管蛋白免疫组织化学表达与黏膜下神经丛形态在不同神经支配段先天性巨结肠（HD）和全结肠无神经节细胞症（TCA）中的关系。
共对 25 例（22 例 HD，3 例 TCA）和 18 例对照进行钙视网膜蛋白和β-微管蛋白免疫组织化学检测。通过视觉分级评分评估代表远段无神经节、过渡段和近段神经节段的切片；β-微管蛋白也通过图像分析进行评估。通过细胞形态学测量黏膜下神经丛的肥大和增生。近端节段的长度与术后肠功能相关。
对照组表现出强烈的钙视网膜蛋白和β-微管蛋白染色。在 HD 和 TCA 中，钙视网膜蛋白染色与神经节细胞的存在有关：远段为阴性，过渡段为弱阳性，近段为强阳性。β-微管蛋白染色在 HD 的所有节段均较弱，在 TCA 中为阴性。肥大和增生的神经丛特征性地存在于无神经节段，在过渡段和神经节段观察到神经逐渐变小。59%的 HD 患者术后出现短暂性便秘、大便失禁或结肠炎，但与近端节段长度或肥大神经丛的存在无明显关系。
钙视网膜蛋白是神经节细胞存在的可靠标志物，与神经肥大一起有助于识别过渡区。切除标本近端节段的长度和神经大小并不影响术后肠道功能。整个结肠段（包括近端神经节段）β-微管蛋白表达减少可能是肠神经传递的潜在损害因素。
