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目的筛选并鉴定Alport综合征(AS)患者与健康对照者(NC)差异性表达蛋白质,寻找AS的生物标记物。方法收集AS组与NC组的尿液,从尿液中分离尿肾脏管细胞,将尿肾脏管细胞诱导分化成多潜能干细胞(i PSCs)。运用i TRAQ技术找出两组之间差异性表达蛋白质。采用Western blot检测差异蛋白质。结果共鉴定出12 600条特有肽段序列,对应3 470种非冗余蛋白质。在两组样本中发现383种蛋白质具有差异性表达,其中差异2倍以上的蛋白质有35种,包括18种上调蛋白质和17种下调蛋白质。Western blot检测KRT14、TUBA1A、PAPSS1、UTF1、SF3B14、MTHFD2等6种差异蛋白质在各组中的表达结果与i TRAQ检测结果一致。结论筛选得到与AS发病相关的差异蛋白质,可以作为AS早期诊断和治疗的生物标记物。
Objective To screen and identify differentially expressed proteins in patients with Alport syndrome (AS) and healthy controls (NCs) for biomarkers of AS. Methods Urine was collected from AS group and NC group. Urinary renal tubular cells were isolated from urine and induced to differentiate into renal tubular cells (iPSCs). Use i TRAQ technology to identify differentially expressed proteins between the two groups. Western blot was used to detect differential proteins. Results A total of 12 600 unique peptide sequences were identified, corresponding to 3 470 non-redundant proteins. 383 proteins were found differentially expressed in two samples, of which 35 were more than twice as many, including 18 up-regulated proteins and 17 down-regulated proteins. Western blot detection of KRT14, TUBA1A, PAPSS1, UTF1, SF3B14, MTHFD2 and other six kinds of differential proteins in each group in the expression results and i TRAQ test results. Conclusion The differential proteins related to the pathogenesis of AS are screened and can be used as biomarkers for the early diagnosis and treatment of AS.