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使用纳米级液相色谱串联质谱分析法对原发性膜性肾病进行尿液蛋白质组学研究。

Urine proteomics of primary membranous nephropathy using nanoscale liquid chromatography tandem mass spectrometry analysis.

作者信息

Pang Lu, Li Qianqian, Li Yan, Liu Yi, Duan Nan, Li Haixia

机构信息

1Department of Clinical Laboratory, Peking University First Hospital, Beijing, China.

2Laboratory of Interdisciplinary Research, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.

出版信息

Clin Proteomics. 2018 Feb 7;15:5. doi: 10.1186/s12014-018-9183-3. eCollection 2018.

DOI:10.1186/s12014-018-9183-3
PMID:29445323
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5801694/
Abstract

BACKGROUND

Primary membranous nephropathy (PMN) is an important cause of nephrotic syndrome in adults. Urine proteome may provide important clues of pathophysiological mechanisms in PMN. In the current study, we analyzed and compared the proteome of urine from patients with PMN and normal controls.

METHODS

We performed two technical replicates (TMT1 and TMT2) to analyze and compare the urine proteome from patients with PMN and normal controls by tandem mass tag (TMT) technology coupled with nanoscale liquid chromatography tandem mass spectrometry analysis (LC-MS/MS). Gene ontology (GO) enrichment analysis was performed to analyse general characterization of the proteins. The proteins were also matched against the database of Kyoto Encyclopedia of Genes and Genomes (KEGG). For validation, Western blot was used to analyze the selected proteins.

RESULTS

A total of 509 proteins and 411 proteins were identified in TMT1 and TMT2, respectively. 249 proteins were both identified in two technical replicates. GO analysis and KEGG analysis revealed immunization and coagulation were predominantly involved. Among the differential protein, the overexcretion of alpha-1-antitrypsin (A1AT) and afamin (AFM) were validated by Western blot analysis.

CONCLUSIONS

Our data showed the important role of immunologic mechanism in the development of PMN, and the value of urinary A1AT and AFM in biomarker discovery of patients with PMN. The discovery of the overexcretion of A1AT and AFM in the urine can help to further elucidate pathogenetic mechanisms involved in PMN.

摘要

背景

原发性膜性肾病（PMN）是成人肾病综合征的重要病因。尿液蛋白质组可能为PMN的病理生理机制提供重要线索。在本研究中，我们分析并比较了PMN患者和正常对照者的尿液蛋白质组。

方法

我们进行了两次技术重复（TMT1和TMT2），通过串联质谱标签（TMT）技术结合纳米液相色谱串联质谱分析（LC-MS/MS）来分析和比较PMN患者和正常对照者的尿液蛋白质组。进行基因本体（GO）富集分析以分析蛋白质的一般特征。这些蛋白质还与京都基因与基因组百科全书（KEGG）数据库进行比对。为进行验证，采用蛋白质印迹法分析所选蛋白质。

结果

在TMT1和TMT2中分别鉴定出509种蛋白质和411种蛋白质。在两次技术重复中均鉴定出249种蛋白质。GO分析和KEGG分析显示主要涉及免疫和凝血过程。在差异蛋白质中，α-1抗胰蛋白酶（A1AT）和载脂蛋白A（AFM）的过度排泄通过蛋白质印迹分析得到验证。

结论

我们的数据显示了免疫机制在PMN发病中的重要作用，以及尿A1AT和AFM在PMN患者生物标志物发现中的价值。尿液中A1AT和AFM过度排泄的发现有助于进一步阐明PMN的发病机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/81242a63fa9a/12014_2018_9183_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/d4819818bb80/12014_2018_9183_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/e740a0747f6a/12014_2018_9183_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/57f62bbb72a3/12014_2018_9183_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/601e70ee793c/12014_2018_9183_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/7ede6558f607/12014_2018_9183_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/37c72f2ded92/12014_2018_9183_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/a0f0d821410f/12014_2018_9183_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/a3b16a29ef33/12014_2018_9183_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/81242a63fa9a/12014_2018_9183_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/d4819818bb80/12014_2018_9183_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/e740a0747f6a/12014_2018_9183_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/57f62bbb72a3/12014_2018_9183_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/601e70ee793c/12014_2018_9183_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/7ede6558f607/12014_2018_9183_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/37c72f2ded92/12014_2018_9183_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/a0f0d821410f/12014_2018_9183_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/a3b16a29ef33/12014_2018_9183_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5abb/5801694/81242a63fa9a/12014_2018_9183_Fig9_HTML.jpg

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