先天性巨结肠c-kit^+肠间质细胞分布的免疫组织化学研究

Immunohistochemical study of distribution of c-kit positive interstitial cells of Cajal in Hirschsprung's disease

目的 了解先天性巨结肠症 (HD)肠壁内神经节细胞与肠间质细胞 (ICCs)分布的关系 ,以进一步研究HD的发病机制。方法 采用兔抗人多克隆c kit抗体 ,通过免疫组化SABC法观察了5 8例HD患儿及 12例正常儿结、直肠壁内c kit+ ICCs的分布情况。患儿年龄 3个月～ 10岁。男 4 6例 ,女 12例。短段型HD 7例 ,常见型 30例 ,长段型 16例 ,全结肠型 5例。结果 对照组肠肌层内及肌间神经节周围分布有中等量至大量的c kit+ ICCs ,并连接成网络状结构 ,而在肌间神经节和粘膜下神经节内均无c kit免疫反应性表达。在HD有神经节细胞段肠壁内c kit+ ICCs的数目和分布与正常对照组无显著差异 (P >0 .0 5 )。而在HD无神经节细胞段肠壁肌层及肌间神经节周围c kit+ICCs的数目明显减少甚至消失 ,与HD有神经节细胞段及正常对照组相比 ,差异有显著性意义 (P <0 .0 1)。结论 c kit+ ICCs的异常分布可能与HD肠动力障碍的发生机制有关。

Objective To investigate the relationship between distribution of the ganglionic cells and interstitial cells of Cajal (ICCs) in the colonic and rectal wall of Hirschsprung's Disease(HD). Methods Fifty eight cases (male 46, female 12 ) aging from 3 months to 10 years of HD were reported. Of which, short segment type was in 7 cases, common type in 30, long segment 16 and total colonic type 5. The distribution of ICCs in between the muscle layers of bowel from normal controls (12 cases) and HD patients was studied by c kit immunohistochemistry. Antihuman c kit serum was used for visualization of c kit positive (c kit +)cells. Results In bowels from normal controls and the normal segment of bowels from patients with HD, moderate to large amount of c kit + ICCs were observed in between the muscle layers. Myenteric plexuses were clearly demarcated by c kit + cells. C kit + ICCs in both the muscle layers and around the myenteric plexus appeared as a network structure, but c kit immunoreactivity did not stain elements within the myenteric and submucosal ganglia. On the contrary, the number of c kit + ICCs was remarkably reduced or even disappeared in between the muscle layers of aganglionic segment of bowel in HD. Statistics showed the significant difference of c kit + ICCs between the aganglionic and ganglionic bowel segments ( P < 0.01 ), but there was no significant difference in the number of c kit + ICCs between the ganglionic segment of bowels in HD and that of normal controls ( P > 0.05 ). Conclusion These results suggest that lack of c kit + ICCs, an intestinal pacemaker, may lead to the bowel spasm in HD.