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运用液相色谱-串联质谱法对儿童过敏性紫癜性肾炎进行尿液蛋白质组学研究

Urinary proteomics of Henoch-Schönlein purpura nephritis in children using liquid chromatography-tandem mass spectrometry.

作者信息

Fang Xiang, Wu Heyan, Lu Mei, Cao Yan, Wang Ren, Wang Meiqiu, Gao Chunlin, Xia Zhengkun

机构信息

1Department of Pediatrics, Jinling Hospital, the First School of Clinical Medicine, Southern Medical University, No. 305 Zhongshan East Road, Nanjing, 210002 Jiangsu China.

Department of Clinical Medicine, Anqing Medical College, Anqing, 246052 Anhui China.

出版信息

Clin Proteomics. 2020 Mar 12;17:10. doi: 10.1186/s12014-020-09274-x. eCollection 2020.

Abstract

BACKGROUND

Henoch-Schönlein purpura nephritis (HSPN) is the principal cause of morbidity and mortality in children with Henoch-Schönlein purpura (HSP). However, the criteria for risk assessment currently used is not satisfactory. The urine proteome may provide important clues to indicate the development of HSPN.

METHODS

Here, we detected and compared the urine proteome of patients with HSPN and healthy controls by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the data-independent acquisition (DIA) mode. The differentially expressed proteins were analysed by gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. For validation, enzyme-linked immunosorbent assay (ELISA) was used to analyse the selected proteins.

RESULTS

A total of 125 proteins (29 upregulated and 96 downregulated) were found to be differentially expressed in children with HSPN compared with the controls. Forty-one proteins were predicted to have direct interactions. The enriched pathways mainly included focal adhesion, cell adhesion molecules, the PI3K-Akt signalling pathway, ECM-receptor interactions and so on. Cell adhesion related to the pathogenesis of HSPN was the main biological process identified in this study. The decrease in two proteins (integrin beta-1 and tenascin) was validated by ELISA.

CONCLUSIONS

Our study provides new insights into the assessment of HSPN progression in children, as well as new potential biomarkers. The data confirm the value of the urinary proteome in capturing the emergence of HSPN.

摘要

背景

过敏性紫癜肾炎(HSPN)是过敏性紫癜(HSP)患儿发病和死亡的主要原因。然而,目前使用的风险评估标准并不令人满意。尿液蛋白质组可能为HSPN的发展提供重要线索。

方法

在此,我们采用液相色谱-串联质谱(LC-MS/MS)的数据非依赖采集(DIA)模式,检测并比较了HSPN患者和健康对照者的尿液蛋白质组。通过基因本体(GO)分析和京都基因与基因组百科全书(KEGG)分析对差异表达蛋白进行分析。为进行验证,采用酶联免疫吸附测定(ELISA)分析所选蛋白。

结果

与对照组相比,共发现125种蛋白(29种上调和96种下调)在HSPN患儿中差异表达。预测有41种蛋白存在直接相互作用。富集的通路主要包括粘着斑、细胞粘附分子、PI3K-Akt信号通路、细胞外基质-受体相互作用等。与HSPN发病机制相关的细胞粘附是本研究确定的主要生物学过程。通过ELISA验证了两种蛋白(整合素β-1和腱生蛋白)的减少。

结论

我们的研究为评估儿童HSPN的进展提供了新的见解,以及新的潜在生物标志物。数据证实了尿液蛋白质组在捕捉HSPN出现方面的价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab4d/7066733/cb605658b4a2/12014_2020_9274_Fig1_HTML.jpg

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