Zhang Shu-Cheng, Chen Fang, Jiang Kai-Lei, Yuan Zheng-Wei, Wang Wei-Lin
Department of Pediatric Surgery, Major Laboratory of Chinese Health Ministry for Congenital Malformations, Shengjing Hospital of China Medical University, Shenyang, China.
Department of Hematology, Shengjing Hospital of China Medical University, Shenyang, China.
Pediatr Res. 2014 Jun;75(6):754-61. doi: 10.1038/pr.2014.33. Epub 2014 Apr 7.
Hirschsprung disease (HSCR) is the third most common congenital disorder of the gastrointestinal tract. This study aims to elucidate changes in protein expression between the normal and aganglionic hindgut in human HSCR.
The biopsies were obtained from the normal and aganglionic hindgut in human HSCR, and the comparative proteomics were analyzed by mass spectrometry (MS)-based two-dimensional gel electrophoresis (2DE).
A total of 932-986 protein spots were identified in each of the gut segments, among which 30 spots had at least an eightfold difference in volume (%). Of the 30 differentially expressed spots, 15 proteins were identified via sequence analysis. Among these 15 proteins, eight were upregulated and seven were downregulated in the aganglionic group. The well-represented classes included biomarkers of enteric ganglions, extracellular matrix proteins, LIM domain proteins, serum proteins, and other pleiotropic proteins. Five proteins were selected and verified by western blotting and real-time PCR, and the results were consistent with the results of 2DE.
MS-based 2DE can help to identify pathological relevant proteins in HSCR; it defines an extensive protein catalog of the normal and aganglionic hindgut and may constitute the basis to understand pathophysiological mechanisms related to the HSCR.
先天性巨结肠（HSCR）是第三常见的先天性胃肠道疾病。本研究旨在阐明人类HSCR中正常与无神经节的后肠之间蛋白质表达的变化。
从人类HSCR的正常和无神经节后肠获取活检组织，通过基于质谱（MS）的二维凝胶电泳（2DE）分析比较蛋白质组学。
在每个肠段中总共鉴定出932 - 986个蛋白质斑点，其中30个斑点的体积（%）至少有八倍差异。在这30个差异表达的斑点中，通过序列分析鉴定出15种蛋白质。在这15种蛋白质中，无神经节组中有8种上调，7种下调。代表性的类别包括肠神经节的生物标志物、细胞外基质蛋白、LIM结构域蛋白、血清蛋白和其他多效性蛋白。选择5种蛋白质通过蛋白质免疫印迹和实时PCR进行验证，结果与2DE结果一致。
基于MS的2DE有助于鉴定HSCR中与病理相关的蛋白质；它定义了正常和无神经节后肠的广泛蛋白质目录，并可能构成理解与HSCR相关的病理生理机制的基础。
