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目的 本研究旨在探讨先天性巨结肠(Hirschsprung’s disease,HD)肠壁中Smoothelin(SM)的变化和临床意义. 方法 自2007年1月至2013年1月,随机选取53例经病理证实先天性巨结肠患儿术中切除的肠段样本(狭窄段即无神经节细胞段、移行段和扩张段即有神经节细胞段).男33例,女20例,年龄3月龄~4岁(平均年龄1.3岁).取材分别为狭窄段、移行段和扩张段肠壁组织各1.0cm3大小,迅速用pH7.4的PBS液冲洗,不同浓度的蔗糖溶液中脱水,-80℃保存.同时留取相同部位的肠壁组织做组织HE染色病理学检查外,辅以PGP9.5、神经元特异性烯醇化酶(NSE)及S-100免疫组织化学染色,证实符合HD诊断.采用RealTime-PCR和免疫蛋白印记法方法从RNA和蛋白水平对SM在HD患儿肠道组织内的表达进行检测,并分析其参与HD发病过程的可能机制. 结果 SM mRNA表达在先天性巨结肠病扩张段、移行段和狭窄段分别为(27.13±16.44)×10-3、(21.60±10.02)×10-3和(19.66±9.92)×10-3;蛋白表达分别为43.13±12.44、36.83±9.43、23.63±4.97.移行段和狭窄段SM mRNA和蛋白均低于扩张段(P<0.05).结论 SM可能参与HD的发病过程.“,”Objective To explore the expression level of smoothelin (SM) protein and mRNA in Hirschsprung’s disease (HD) and elucidate the relationship between the expression of SM and the pathogenesis of HD.Methods Full-thickness colonic specimens were obtained from 53 HD patients undergoing pull-through procedure between January 2007 and January 2013.There were 33 boys and 20 girls with an age range of 5 months to 4 years.The diagnosis of HD was con?rmed jointly by hematoxylin & eosin histochemistry,PGP 9.5,neuron specific enolase (NSE) and S100 immunohistochemistry.Quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used for detecting SM mRNA and protein expression in different segments of colonic tissue.Results The expression levels of SM mRNA and protein were 27.13±16.44×10-3,21.60±10.02×10-3,19.66±9.92×10-3 and 43.13±12.44,36.83±9.43,23.63±4.97 in ganglionic,transitional and aganglionic segments respectively.Both mRNA and protein of SM were markedly less in transitional and aganglionic segments than those in ganglionic segment (P<0.05).Conclusions SM may be involved in the pathogenesis of HD.