慢传输型便秘大鼠结肠黏膜下调蛋白质组的分离鉴定及其功能分析

Separation and identification of down-regulated proteomics of colonic mucosa of chronic slow transit constipation rat

目的研究慢传输型便秘大鼠结肠黏膜下调蛋白质的整体变化特性,并对明确的蛋白质进行功能分析,为探讨慢传输型便秘大鼠结肠黏膜损害的发病机制提供理论依据。方法利用复方苯乙哌啶建立大鼠慢传输型便秘模型,利用高分辨双向电泳(2-DE)对STC大鼠结肠黏膜组织进行蛋白质分离,ImageMaster 2D Elite图像分析软件进行分析,应用生物质谱、蛋白质库及文献分析等技术对差异蛋白质中的下调蛋白质组进行鉴定、功能和临床意义分析。结果慢传输型便秘在其发生过程中有显著的蛋白质表达差异。主要差异表达的蛋白质有肥大细胞蛋白酶(A1)、非特异性二肽酶(A2)、线粒体脱氢酶前体(A3),A1、A2、A3在便秘大鼠的胶图中表达明显下调。结论这些下调的蛋白质的表达改变提示慢传输型便秘大鼠结肠组织免疫反应性减弱,线粒体氧化还原功能障碍在慢传输型便秘的发生、发展中可能起重要作用,慢传输型便秘大鼠肠道蛋白质的消化、吸收功能可能存在一定程度的障碍。

Objective To investigate the general changes of down-regulated proteomics of colonic mucosa of chronic slow transit constipation rat and analyze the function of the specified proteomics so as to provide theoretical basis for the pathogenesis of damage to intestinal mucosa of chronic slow transit constipation rat. Methods Establish slow transit constipation model in rats by Diphenoxylate. An experimental model of chronic slow transit constipation rat was used for separation of proteomics from colonic mucosa by using two-dimensional electrophoresis （2DE） . ImageMaster 2DElite , mass spectrometry , protein bank and bibliometrics were applied to identify the down-regulated proteomics in the difference proteins and analyze their clinical significance and function. Results Variances of protein expression of rat colon under different conditions were analyzed by two-dimensional gel electrophoresis. Obvious distinction was observed in the genesis of STC, such as follows： mast cell protease（A1）, non-specific dipeptidase （A2）, mitochondrial aldehyde dehydrogenase precursor（A3 ）. The expression of A1, A2, A3 down-regulated in the gel graph of STC rats. Conclusion The down-regulation of mitochondrial aldehyde dehydrogenase,mast cell protease as well as non-specific dipeptid- ase in rat colon suggests the functional impairment of the oxidoreduction of mitochondrion are very important in the genesis and development of STC. The immunological reaction of STC rats weakens, the function of digesting and absorbing protein may be damaged to some extent.