利用基质辅助激光解析电离飞行时间串联质谱分析胆囊结石中非黏性蛋白的组成
发布时间：2018-09-04 14:42
【摘要】：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。
[Abstract]:Objective to investigate the presence of non-viscosity protein in cholesterol gallstone and pigment gallstone and to identify the protein composition. Methods 30 patients (11 males and 19 females) underwent cholecystolithiasis. Gallstones were classified into 20 cholesterol type and 10 bile pigment type by naked eye observation and triglyceride detection kit (GPO-PAP). Gallstone protein was extracted by tissue lysis method (RIPA) and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) combined with matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-TOF). Analysis of gene bulk (GO) function enrichment by DAVID tools. This map and Kyoto gene, genome encyclopedia (KEGG) signal transduction pathway enrichment analysis. Results specific proteins were found in cholesterol gallstones and pigment gallstones, which were identified as transferrin precursor protein (TRFE), albumin (ALB), antitrypsin 1 (SERPINA1), immunoglobulin 纬 -1 (IGHG1), 伪 hemoglobin (HBA1) and 尾 hemoglobin (HBB), in which IGHG1 was not present in gallstones. Conclusion TRFE,ALB,SERPINA1,HBA1,HBB and other proteins co-exist in cholesterol and pigment gallstone, while IGHG1 exists only in cholesterol stone. IGHG1 protein or its corresponding gene is an important element and biomarker. KEGG analysis shows that SERPINA1 is involved in complement and coagulation cascade reaction. It is important to identify and analyze the components of non-mucin in gallstone and use the simulation of database to elucidate the pathogenesis of gallstone.
【作者单位】： 贵阳医学院;厦门大学药学院;贵阳医学院附属医院肝胆外科;
【基金】：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)
【分类号】：R575.62
[Abstract]:Objective to investigate the presence of non-viscosity protein in cholesterol gallstone and pigment gallstone and to identify the protein composition. Methods 30 patients (11 males and 19 females) underwent cholecystolithiasis. Gallstones were classified into 20 cholesterol type and 10 bile pigment type by naked eye observation and triglyceride detection kit (GPO-PAP). Gallstone protein was extracted by tissue lysis method (RIPA) and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) combined with matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-TOF). Analysis of gene bulk (GO) function enrichment by DAVID tools. This map and Kyoto gene, genome encyclopedia (KEGG) signal transduction pathway enrichment analysis. Results specific proteins were found in cholesterol gallstones and pigment gallstones, which were identified as transferrin precursor protein (TRFE), albumin (ALB), antitrypsin 1 (SERPINA1), immunoglobulin 纬 -1 (IGHG1), 伪 hemoglobin (HBA1) and 尾 hemoglobin (HBB), in which IGHG1 was not present in gallstones. Conclusion TRFE,ALB,SERPINA1,HBA1,HBB and other proteins co-exist in cholesterol and pigment gallstone, while IGHG1 exists only in cholesterol stone. IGHG1 protein or its corresponding gene is an important element and biomarker. KEGG analysis shows that SERPINA1 is involved in complement and coagulation cascade reaction. It is important to identify and analyze the components of non-mucin in gallstone and use the simulation of database to elucidate the pathogenesis of gallstone.
【作者单位】： 贵阳医学院;厦门大学药学院;贵阳医学院附属医院肝胆外科;
【基金】：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)
【分类号】：R575.62
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