亚健康大便不成形人群结肠黏膜蛋白质组学研究

Proteomics of the colonic mucosa in sub-healthy people with shapeless stool

目的:利用蛋白质组学方法建立亚健康大便不成形人群结肠黏膜蛋白质图谱,并对部分表达明显差异的蛋白质进行鉴定,为探讨亚健康大便不成形人群的结肠黏膜改变机制提供理论依据。方法:采用高分辨双向凝胶电泳(2-DE)对亚健康大便不成形人群及健康志愿者结肠黏膜组织进行蛋白质分离,Im-ageMaster 2D Elite分析软件进行图像分析,应用生物质谱、蛋白质数据库及信息技术对表达明显差异的蛋白质进行质谱鉴定分析。结果:大便不成形组平均胶蛋白斑点数为517,正常对照组平均胶蛋白斑点数为535,与正常对照组平均胶蛋白斑点匹配数为366,匹配率为70.79%。对10个表达量存在显著差异的蛋白质点进行质谱分析鉴定出8个蛋白质,其中5个蛋白表达下调,3个蛋白表达上调。结论:亚健康大便不成形人群和正常人的结肠黏膜组织蛋白质表达存在差异,醛脱氢酶A1、线粒体羟甲基辅酶A合成酶、γ-肌动蛋白及膜联蛋白A5等8种蛋白可能与亚健康大便不成形的发生相关。

Objective To establish the 2-dimensional electrophoresis （2-DE） map in colonic mucosa in sub-heahhy people with shapeless stool and healthy peopIe, to identify the differential proteins by matrix-assisted laser desorption/ionization time of flight mass spectrometry （ MALDI- TOF-MS）, and to provide theoretical basis for the pathogenesis of intestinal mucosa in sub-healthy people with shapeless stool. Methods Two-DE was used to separate the total proteins from the in- testinal mucosa in sub-heahhy people （the sub-health group） with the shapeless stool and healthy volunteers （the control group）. ImageMaster 2D Elite soft was applied to analyze the 2-DE images, and the differentially expressed protein spots between the 2 groups were identified by MALDI-TOF- MS, protein bank and information technique. Results We analyzed the average maps and obtained 517 protein spots in the sub-healthy group and 535 protein spots in the control group. Between the sub-healthy group and the control group, the mean of 366 protein spots was matched, and the matching rate was 70.79%. Ten differential protein spots were screened by MALDI-TOF-MS, and 8 were identified. Five out of the 8 spots were significantly decreased, while 3 out of the 8 were significantly increased. Conclusion The proteomic expression in colonic mucosa of people with shapeless stool is significantly different from that of healthy people. Eight differential proteins such as aldehyde dehydrogenase 1A1 isoform 1, 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2 （mi- tochondrial） ,γ-actin, annexin A5 possibly involve in the pathogenesis of sub-healthy people with shapeless stool.