荧光差异电泳和质谱技术联合研究对比剂肾损伤尿液差异表达蛋白
Differentially expressed urine protein of contrast induced acute kidney injury by two-dimensional differential in-gel electrophoresis and mass spectrometry
摘要目的 比较对比剂肾损伤发生前后尿液蛋白质组的差异,搜寻差异表达蛋白,为对比剂肾病发病机制的阐明和尿液标志蛋白的发现提供依据.方法 收集12例上海交通大学医学院附属仁济医院心内科接受冠状动脉造影或介入诊疗术患者手术前、后的尿液,用不同的荧光染料标记后行双向电泳,Decyder软件比较蛋白质谱的差异,对差异表达点用质谱技术进行蛋白质鉴定.结果 发现了56个差异表达蛋白点,其中39个点上调,17个点下调.其中,凝集素补体激活途径的关键蛋白-甘露糖结合凝集素和甘露糖结合凝集素相关的丝氨酸蛋白酶2均上调.结论 建立了基于荧光差异电泳和质谱分析技术的尿液蛋白质组学方法,搜寻了对比剂急性肾损伤发生前后的差异表达蛋白质,提示甘露糖结合凝集素补体激活途径可能参与了对比剂急性肾损伤的发病.
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abstractsObjective To explore the differentially expressed proteins of contrast-induced nephropathy through a comparison of urinary proteome so as to further elucidate the pathogenesis and discover the disease biomarker.Methods The urine samples of 12 patients were collected before and after coronary angiography.Two dimensional electrophoresis was performed after the urine samples were labeled by different dyes.The differences of urine proteome were analyzed by Decyder software and the differentially expressed spots identified by mass spectrometry.Results A total of 56 differentially expressed spots were detected.Among them, 39 spots were up-regulated and 17 spots down-regulated.And mannose binding lectin and mannose binding lectin associated serine protease 2, key proteins in complement body activation, were both significantly up-regulated.Conclusion Urine proteomic study methods are constructed based on 2D-DIGE and mass spectrometry.The lectin pathway of complement body may be associated with contrast-induced acute kidney injury.
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