先天性巨结肠中HA117及SHH信号通路的表达及意义  

Expression profiles and significance of HA117 and SHH signaling pathway in Hirschsprung＇s disease

作　　者：滕银平 金先庆[1] 李力[1] 吴欢[1] 王鑫[1] 罗媛圆 

机构地区：[1]儿童发育疾病研究教育部重点实验室,儿科学重庆市重点实验室,重庆市儿童发育重大疾病诊治与预防国际合作基地,重庆医科大学附属儿童医院胃肠新生儿外科,400014

出　　处：《中华小儿外科杂志》2016年第4期253-257,共5页Chinese Journal of Pediatric Surgery

基　　金：国家临床重点专科建设项目[国卫办医函（2013）554号]

摘　　要：目的研究HA117及SHH信号通路（SHH、Ptch1、Gli1）在先天性巨结肠（Hirschsprung＇s disease，HD）中的表达情况，以探讨其与HD发生发展中的关系。方法收集重庆医科大学附属儿童医院2014年至2015年经手术确诊为HD患儿的巨结肠组织标本30例，肠段分吻合段（正常对照组）、扩张段和痉挛段（实验组），运用实时荧光定量PCR（quantitative real-time PCR，qRT-PCR）、蛋白质印迹（Western blot）和免疫组织化学方法检测HA117、SHH信号途径在各段中的表达情况，对其进行定量和定性并比较。结果qRT-PCR示痉挛段HA117 mRNA表达水平为1.52±0.17，明显高于吻合段的0.28±0.04，差异具有统计学意义（P〈0.05）。痉挛段SHH、Ptch1、Gli1 mRNA表达水平分别为0.53±0.11、0.48±0.11、0.62±0.13，明显低于吻合段（1.23±0.26、1.20±0.22、1.71±0.46），差异具有统计学意义（P〈0.05）；Western-Blot示SHH、Ptch1、Gli1蛋白相对表达水平与mRNA表达水平一致，差异均具有统计学意义（P〈0.05）；免疫组织化学（DAB染色）结果提示：痉挛段SHH、Ptch1、Gli1染色明显弱于吻合段。结论HA117及SHH信号通路可能与肠神经系统（ENS）发育有着密切的联系，这可能是HD病理发生的重要作用机制之一。Objective To explore the relationship between HA117 and SHH signaling pathway （SHH, Ptchl ＆ Glil） and Hirschsprung＇s disease （HD） by analyzing their expression profiles in different colonic segments of HD patients. Methods Colonic specimens were collected intraoperatively from 30 patients diagnosed with HD at our hospital from 2014 to 2015. Proximal anastomosis, dilated segment and stenotic segment were designated. The mRNA levels of HA117, SHH, Smo, Ptchl and Glil were evaluated by quantitative real-time polymerase chain reaction （PCR）. And the protein levels of SHH, Ptchl and Glil were examined by Western blot and immunohistochemistry. Results The mRNA levels of HA117 were significantly higher in stenotic segment than those in proximal anastomosis（1.52 ± 0. 17 vs 0. 28 ± 0. 04, P〈0. 05）. And the mRNA levels of SHH, Ptchl and Glil were significantly lower in stenotic segment than those in proximal anastomosis （0. 53 ± 0. 11, 0. 48 ± 0. 11 ＆ 0. 62 ± 0. 13 vs t. 23 ±0. 26, 1.20 ± 0. 22 ＆ 1.71 ± 0. 46, P〈0. 05）. The protein levels of SHH, Ptchl and Glil were consistent with their mRNA levels （P〈 0. 05）. Immunohistochemical staining of SHH, Ptchl and Glil were significantly less intense in stenotic segment than those in proximal anastomosis. Conclusions HA117 and SHH signaling pathway may have close connections with the development of enteric nervous system （ENS）. And it may be one of the important mechanisms of activating the pathogenesis of HD.

关 键 词：先天性巨结肠 肠神经系统 神经元 

分 类 号：R726.5[医药卫生—儿科]