先天性巨结肠HuD蛋白表达的实验研究

目的 HuD是神经元胚胎发育中生长锥、突起形成延伸及成熟神经元存活所必须的蛋白，其在肠神经系统发育及先天性巨结肠发生中作用机制尚不明确，国内外均未见相关报道。本文探讨HuD蛋白在先天性巨结肠不同部位肠壁组织中的表达，以了解HuD在先天性巨结肠发病中作用机制。方法分别取15例先天性巨结肠患儿手术切除的病变段、移行段、扩张段结肠壁全层及5例人正常结肠组织，采用免疫组织化学、Western免疫印迹分析对不同部位肠壁HuD蛋白表达进行检测，利用图象分析系统及统计软件进行结果分析。结果先天性巨结肠狭窄段粘膜下和肌间抗HuD蛋白抗体染色缺如，移行段显色程度及表达量都明显减少于扩张段和正常对照组（P〈0．01）；狭窄段和移行段的HuD蛋白水平明显少于扩张段和正常段（P〈O．01）。结论HuD在先天性巨结肠病变段粘膜下、肌间神经丛不表达提示HuD蛋白减少可能是使肠神经系统胚胎发育或成熟神经元存活出现障碍而导致先天性肠无神经节细胞的原因之一。

Study on The Expression of HuD protein in Hirschsprung's Disease

Objective HuD protein involved in neurite growth and mature neurons survival is the necessary factor in the embryonic and adult developing nervous system. It is not clearly that the role of HuD in developing enteric nervous system and pathogenesis of Hirschsprung's disease. There is no allied reports both at home and abroad. This study about the expression of HuD in the different bowel segment of Hirschsprung's disease, is maybe help for us to understand the relationship between HuD and Hirschsprung's disease. Methods To obtain the different bowel segment ablated in operation from 15 cases of Hirschsprung's disease and the colon with normal ganglion ablated operation by other causes. Using immuocytochemistry and western blotting to determine the expression of HuD in the different bowel segment of Hirschsprung's disease. To analysis the result by image dissector system and statistic software. Results The stain of anti-HuD protein was negative both the submucosa of trictural segment and intermuscles.The expression degree and stain of transitional segment were all low than that of both dilated segment and normal control group(P<0.01).Hud protein level in strictural and transitional segment remarkable low than that in dilated and normal segments(P<0.05) in Hirschsprug's disease. Conclusions HuD in the submucosa and intermuscular nervous plexus was no expressed,it suggest that HuD protein decrease could cause enonervous system embryodevelopment or mature neuron surival disorder.Finalyty,these disorders, lead to congenital aganglion cells in bowel of patients with Hirschsprung,s disease.