目的探讨胆固醇结石和胆色素结石中是否存在非黏性蛋白并鉴定蛋白质的组成。方法选取30例行保胆取石术患者的胆囊结石,男性11例,女性19例。采用肉眼观察并结合三酰甘油检测试剂盒(gPO-PAP)测定胆固醇含量的方法对胆结石进行分类,分为20例胆固醇型和10例胆色素型。应用组织裂解法(rIPA)提取胆囊结石蛋白,并用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SdS-PAgE)与基质辅助激光解析电离飞行时间串联质谱(MAldI-TOf-TOf)分析相结合的方法对该蛋白进行鉴定分析。通过dAVId工具对上述基因进行基因本体(gO)功能富集分析本图及京都基因、基因组百科全书(kEgg)信号转导通路富集分析。结果胆固醇结石和胆色素结石中出现特异性蛋白,经质谱鉴定为转铁蛋白前体蛋白(TrfE)、白蛋白(Alb)、抗胰蛋白酶1(SErPInA1)、免疫球蛋白γ-1(IgHg1)、α血红蛋白(HbA1)、β血红蛋白(Hbb),其中IgHg1不存在于胆色素结石中。结论 TrfE、Alb、SErPInA1、HbA1、Hbb等蛋白共同存在于胆固醇和胆色素结石中,而IgHg1只存在于胆固醇结石中,可见IgHg1蛋白或其对应基因是参与不同胆结石类型成因的重要元素及生物标志物。kEgg分析结果表明,SErPInA1参与补体和凝血级联反应。鉴定和分析胆囊结石中非黏性蛋白的成分及利用资料库的模拟对阐明胆囊结石的发病机制有重要的意义。【Objective】 To examine the presence of non-mucin proteins in the cholesterol calculi and pigment gallstones, and to further identify the type of proteins.【Methods】 Gallstones were obtained from 30 patients at endoscopic minimally invasive cholecystolithotomy, including 11 males and 19 females.The classification of gallstones was applied by visual observation and measurement of concentration of cholesterol through GPO-PAP method.There were 20 cases of cholesterol gallstones and 10 cases of pigment gallstones.The proteins in gallstones were extracted by RIPA, separated by SDS-PAGE and deciphered by matrix-assisted laser desorption/ ionization time of flight(MALDI-Tof-Tof) mass spectrometry.DAVID was used to analyze the GO(gene ontology) function annotation and the KEGG signal pathway of the targeted genes.【Results】Non-mucin proteins, transferrin(TRFE) precursor protein, ALB, SERPINA1, HBA1 and HBB were found and identified in cholesterol stones and pigment stones; while IGHG1 was only observed in cholesterol stones.【Conclusions】 The six proteins have been first figured out to exist in human gallstones.Interestingly, IGHG1 and its associated genes, solely present in cholesterol gallstones may play a novel role in the classification and/or development of gallstone as a biomarker.In KEGG pathway, SERPINA1 is involved in pathway of complement and coagulation cascades.It is important to further study the origin and the functions of these non-mucin proteins for understanding the pathogenesis of gallstones.贵州省省长基金临床应用课题专项研究项目[No:黔省专合字(2012)94号]; 国家国际科技合作项目(No:2014DFA31420
