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目的搜索常染色体显性多囊肾病(ADPKD)患者与正常成人尿液蛋白质组中有丰度差异的蛋白质成分。方法制备ADPKD患者(PKD1突变)及正常成人尿液蛋白组样品,同位素编码的亲和力标签法标记蛋白质、筛选被标记肽段,二维液相色谱-串联质谱法分离鉴定被标记肽段,从而鉴定蛋白质。通过计算两同位素试剂标记的相同肽段指纹峰面积比,寻找有丰度差异的蛋白质。根据被鉴定肽段的可信度,从鉴定出的蛋白质中选出6种,免疫印迹法验证其在两尿液蛋白组样品中的丰度差异。结果两尿液蛋白质组共鉴定出尿蛋白质106种, ADPKD患者尿液中水平较正常上调者48种,下调者45种。其中多囊蛋白1、肝细胞生长因子、单核细胞趋化蛋白3及孕激素受体等5种蛋白丰度差异为免疫印迹所证实。结论包括多囊蛋白1在内的多种蛋白质成分在ADPKD患者尿液中丰度发生改变,差异蛋白包括生长因子、凋亡调节蛋白、细胞外基质成分、受体、参与胞浆运输的蛋白质、酶类、细胞信号蛋白、转录因子及转录调节因子等,这些蛋白质信息可能为寻找与ADPKD发病相关的蛋白提供部分实验依据。
Objective To search for proteins with abundant abundances in the proteome of patients with autosomal dominant polycystic kidney disease (ADPKD) and normal adults. Methods ADPKD patients (PKD1 mutation) and normal adult urine proteome samples were prepared. The proteins were labeled by isotope coding affinity tag method. The labeled peptides were screened and the identified peptides were separated and identified by two-dimensional liquid chromatography-tandem mass spectrometry. protein. By calculating the peak area ratio of the same peptide finger labeled by two isotopes, the protein with different abundance was searched for. Six of the identified proteins were selected based on the authenticity of the identified peptides and their abundance differences in the two urine protein samples were verified by immunoblotting. Results Urine proteins were identified in 106 urinary proteomes, and urinary protein was identified in 106 urinary samples of ADPKD patients, 48 of them were up-regulated in urine and 45 of them were down-regulated. Among them, the differences of five protein abundances, such as polycystin 1, hepatocyte growth factor, monocyte chemotactic protein 3 and progesterone receptor, were confirmed by Western blot. Conclusions The abundance of various protein components, including polycystin 1, has been changed in the urine of ADPKD patients. The differential proteins include growth factors, apoptosis regulatory proteins, extracellular matrix components, receptors, proteins involved in cytoplasm transport, Enzymes, cell signaling proteins, transcription factors and transcriptional regulators, etc. These protein information may provide some experimental evidence for finding proteins involved in the pathogenesis of ADPKD.