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血清衍生细胞外囊泡的蛋白质组学分析在强直性脊柱炎患者中的应用。

Proteomic analysis of serum-derived extracellular vesicles in ankylosing spondylitis patients.

机构信息

Department of Rheumatology and Immunology, Guangdong Second Provincial General Hospital, Guangzhou 510317, China.

Department of Rheumatology and Immunology, Guangdong Second Provincial General Hospital, Guangzhou 510317, China; The Second School of Clinical Medicine, Southern Medical University, Guangzhou 510515, China.

出版信息

Int Immunopharmacol. 2020 Oct;87:106773. doi: 10.1016/j.intimp.2020.106773. Epub 2020 Jul 14.

Abstract

BACKGROUND

Ankylosing spondylitis (AS) is a chronic inflammatory disease, whose pathogenesis is still unclear. Many studies show the proteins in extracellular vesicle (EVs) would change regularly in many diseases. The study aims to explore the proteins contents of serum-derived EVs in AS patients.

METHODS

EVs were separated by ExoQuick kit. The protein profiles of AS patients and healthy subjects were analyzed by Label-free-liquid chromatography mass spectrometry (LC-MS/MS) technology. Enzyme-linked immunosorbent assay (ELISA) was used to verify the levels of the differently expressed proteins. Receiver operation characteristic (ROC) curves and bioinformatic analysis were conducted.

RESULTS

Six hundred and ten serum-derived EVs proteins from AS patients were detected. Seventy-three diferentially expressed proteins were found in AS group, compared with healthy subjects. Of these, 31 proteins were up-regulated in AS group, while 42 proteins were down-regulated. ELISA result showed that EVs-derived serum amyloid A-1 (SAA1) was higher in AS group, which was consistent with the Label-free-LC-MS/MS data. ROC curves result revealed that the area under curve (AUC) value of EVs-derived SAA1 for AS was 0.768 (0.652-0.885). Bioinformatic analysis revealed that the differently expressed proteins in AS group were significantly involved in "complement and coagulation cascades", "staphylococcus aureus infection", "systemic lupus erythematosus" and "PI3K-Akt signaling pathway".

CONCLUSIONS

The protein profiles of serum-derived EVs in AS patients and healthy subjects were different. EVs-derived SAA1 may be a potential biomarkes of AS. The function analysis indicated that the differentially expressed proteins may potentially participate in immune response.

摘要

背景

强直性脊柱炎(AS)是一种慢性炎症性疾病,其发病机制尚不清楚。许多研究表明,细胞外囊泡(EVs)中的蛋白质在许多疾病中会发生规律性变化。本研究旨在探索 AS 患者血清来源 EVs 中的蛋白质含量。

方法

采用 ExoQuick 试剂盒分离 EVs。采用无标记液相色谱-质谱联用(LC-MS/MS)技术分析 AS 患者和健康受试者的蛋白质谱。采用酶联免疫吸附试验(ELISA)验证差异表达蛋白的水平。进行接收者操作特征(ROC)曲线和生物信息学分析。

结果

从 AS 患者中检测到 610 种血清衍生 EVs 蛋白。与健康受试者相比,AS 组中发现 73 种差异表达蛋白。其中,31 种蛋白在 AS 组中上调,42 种蛋白下调。ELISA 结果表明,AS 组血清淀粉样蛋白 A-1(SAA1)水平较高,与无标记 LC-MS/MS 数据一致。ROC 曲线结果显示,EVs 衍生 SAA1 对 AS 的曲线下面积(AUC)值为 0.768(0.652-0.885)。生物信息学分析显示,AS 组差异表达蛋白主要参与“补体和凝血级联”、“金黄色葡萄球菌感染”、“系统性红斑狼疮”和“PI3K-Akt 信号通路”。

结论

AS 患者和健康受试者血清来源 EVs 的蛋白质谱不同。EVs 衍生的 SAA1 可能是 AS 的潜在生物标志物。功能分析表明,差异表达蛋白可能潜在参与免疫反应。

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