人胆囊组织蛋白质表达谱的建立

Establishment of protein profiling system for human gallbladder tissue

目的:建立胆固醇结石病人与正常人胆囊组织的蛋白质表达谱,用于比较胆囊组织的蛋白质组学研究,为阐明胆石成因提供实验依据。方法:提取胆固醇结石病人及正常人的胆囊壁组织总蛋白,构建相应的双向电泳表达图谱,应用ImageMaster图像分析软件对双向电泳图谱进行图像分析。结果:两组样本均获得分辨率高、重复性好的双向电泳图谱,胆固醇结石组与正常组病人组内蛋白质平均匹配点数分别为632±61和606±64,各匹配率分别为73.46%和71.49%。组间蛋白质平均匹配点为474,匹配率为70.64%。胆固醇结石病人的28个蛋白点表达上调,7个点表达下调,与正常人间蛋白表达差异明显(P<0.05)。结论:初步建立了胆固醇结石病人与正常人胆囊组织的差异蛋白质表达谱,分析两者间差异蛋白点,为胆石成因研究提供实验基础。

Objective To establish an experimental system for the protein profiling of human gallbladder tissue, thus to perform proteomics study. Methods： Whole cell proteins were extracted from the tissues of the gallbladder wall, which were collected from cholesterol gallstone patients and healthy adults respectively. Two-dimensional polyacrylamidegel eleetrophoresis system was used to detect the differential protein expression.ImageMaster two-dimensional electrophoresis analysis software was used for data analysis. Results： The average matched protein spots of the gallbladder stone group and normal control group were 632±61 and 606±64, respectively, with the matched rates of 73.46% and 71.49% correspondingly. The average number of matched spots iu these 2 groups was 474, with the matched rate of 70.64%. The numbers of up-regulated and down-regulated protein spots in gallbladder stone group were 28 and 7, which was significantly different from those in healthy adults. Conclusions： This study has successfully established a highlyresolved and reproducible protein profiling system of human gallbladder tissue, and provided the experimental basis for the etiology study of gallstone.