人去浆膜层胆囊组织蛋白质表达谱的建立与分析

目的 采用蛋白质组学技术分析胆固醇结石患者与正常人去浆膜层胆囊组织蛋白表达谱的差异,通过质谱技术鉴定差异表达蛋白,为胆石成因提供实验依据.方法 提取胆固醇结石患者和正常人去浆膜层胆囊组织蛋白,构建相应的双向电泳蛋白质表达谱并应用ImageMaster图像分析软件获得差异蛋白点信息,采用质谱鉴定差异表达蛋白质.结果 胆固醇结石患者与正常人去浆膜层胆囊组织差异蛋白质表达谱均获得分辨率高、重复性好的双向电泳图谱,差异表达谱结果显示19个蛋白点在两者之间差异较大,质谱成功鉴定11个,1个重复.其中Tropomyosin 4（TPM4）、Transgelin （sm22）、Transthyretin （TTR）、CyanometRhbl.1、Haptoglobin-related protein precursor、Chain A,the structure of Holo type human Cu,Zn superoxide dismutase在结石组中表达上调,Myosin、Peroxiredoxin 3（Prdx3）、Hypothetical protein、T cell receptor alpha chain在结石组中表达下调.TTR、TPM4、SM22、Prdx3蛋白差异为Western blotting和RT-PCR所验证.结论 初步建立了胆固醇结石患者与正常人去浆膜层胆囊组织的差异蛋白质表达谱,分析、鉴定并验证了差异蛋白点,为胆固醇结石形成的关键调控蛋白质研究提供实验依据.

Establishment and analysis of expression profile of serosa-removed gallbladder tissue protein

Objective To use proteomic technology to analyze difference of serosa-removed gallbladder tissue protein expression profile between cholesterol gallstone patients and normal people and to use mass-spectrometric technique to identify differentially expressed proteins, and to provide experimental evidence for causes of gallstones. Methods The serosa-removed gallbladder tissue protein from cholesterol gallstone patients and normal people were extracted, the corresponding two-dimensional electrophoresis protein expression profiles were constructed and image analysis software Image Master was applied to obtain information of different protein spots, mass-spectrometric technique was used to identify differentially expressed proteins. Results The serosa-removed gallbladder tissue protein expression profile of both cholesterol gallstone patients and normal people obtained two-dimensional electrophoresis patterns with high resolution and reproducibility, and differential expression profiles showed that 19 protein spots vary greatly between the two groups, and 11 of them were successfully identified by mass spectrometry. Among them, Tropomyosin 4 （TPM4）, Transgelin （sm22）, Transthyretin （TTR）, Cyanomet Rhb]. 1, Haptoglobin-related protein precursor and Chain A, the structure of Holo type human Cu, Zn superoxide dismutase showed upward expression in gallstone group, while Myosin, Peroxiredoxin 3 （Prdx3）, Hypothetical protein, T cell receptor alpha chain show downward expression in gallstone group. The protein differences of TTR, TPM4, SM22, Prdx3 were verified by Western Blotting and RT-PCR. Conclusion The serosa-removed gallbladder tissue protein expression profile of cholesterol gallstone patients and normal people are initially established. The differential protein spots are analyzed,identified and verified. It provides experimental evidence for protein research as a key regulator of cholesterol gallstone formation.