摘要
目的:Vogt-Koyanagi-Harada 综合征是一种常见的自身免疫性葡萄膜炎,可导致失明。最近的研究表明,血浆外泌体携带与疾病相关的蛋白质,可以作为生物标志物。在这里,我们旨在使用血浆外泌体的蛋白质组学分析来寻找 Vogt-Koyanagi-Harada 病的候选生物标志物。 方法:从以下各组正常对照和 Vogt-Koyanagi-Harada 患者的血浆中分离外泌体:a)初始炎症发作(活动期),b)治疗一个月后缓解(不稳定期),c)静止期治疗三个月后的阶段(稳定阶段)。使用同量异位标记进行相对和绝对定量,通过质谱法分析各组。功能分析后,通过ELISA验证感兴趣的蛋白质。 结果:在外泌体中鉴定出 463 种蛋白质。 43 个在活动性炎症阶段被上调,包括炎症相关蛋白。 31 个被下调。基因本体和通路分析揭示了与细胞粘附、细胞吞噬作用、细胞骨架运动、白细胞跨内皮细胞迁移和血小板活化相关的差异蛋白。通过 ELISA,证实碳酸酐酶 2 和 Ras 相关蛋白 Rap-1b 在外泌体中的活性阶段比正常对照和固定期更丰富,但在微泡或血浆中则不然。 结论: Vogt-Koyanagi-Harada 患者的血浆外泌体含有许多与炎症程度相关的蛋白质。外泌体中碳酸酐酶2和Ras相关蛋白Rap-1b的水平可用作Vogt-Koyanagi-Harada病活动性炎症的生物标志物。进一步的研究有助于研究 Vogt-Koyanagi-Harada 病的发病机制并确定治疗靶点。
关键词: Vogt-Koyanagi-Harada 综合征、外泌体、微泡、细胞外囊泡、蛋白质组学、生物标志物、自身免疫性葡萄膜炎。
Current Molecular Medicine
Title:Proteomics Profiling of Plasma Exosomes in VKH Patients
Volume: 21 Issue: 8
关键词: Vogt-Koyanagi-Harada 综合征、外泌体、微泡、细胞外囊泡、蛋白质组学、生物标志物、自身免疫性葡萄膜炎。
摘要:
Objectives: Vogt-Koyanagi-Harada syndrome is common autoimmune uveitis that can cause blindness. Recent studies have shown that plasma exosomes carry disease-related proteins that may serve as biomarkers. Here, we aimed to find candidate biomarkers of Vogt-Koyanagi-Harada disease using proteomic analysis of plasma exosomes.
Methods: Exosomes were isolated from the plasma of normal controls and Vogt- Koyanagi-Harada patients in the following groups: a) initial inflammatory attack (active stage), b) remission after one month of treatment (unstable stage), and c) stationary phase after three months of treatment (stable stage). Groups were analyzed by mass spectrometry using isobaric tags for relative and absolute quantitation. After functional analysis, proteins of interest were verified by ELISA.
Results: 463 proteins were identified in the exosomes. Forty-three were upregulated at the active inflammation stage, including inflammation-associated proteins. Thirty-one were downregulated. Gene ontology and pathway analyses revealed differential proteins related to cell adhesion, cell phagocytosis, cytoskeleton movement, leukocyte migration across endothelial cells, and platelet activation. By ELISA, Carbonic anhydrase 2 and Ras-related protein Rap-1b were verified as more plentiful at the active stage compared to the normal control and stationary phase in exosomes, but not, however, in microvesicles or plasma.
Conclusion: Plasma exosomes of Vogt-Koyanagi-Harada patients contain many proteins related to the degree of inflammation. The levels of Carbonic anhydrase 2 and Ras-related protein Rap-1b in exosomes can be used as biomarkers for active inflammation in Vogt-Koyanagi-Harada disease. Further investigation could help study the pathogenesis of Vogt-Koyanagi-Harada disease and identify therapeutic targets.
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Cite this article as:
Proteomics Profiling of Plasma Exosomes in VKH Patients, Current Molecular Medicine 2021; 21 (8) . https://dx.doi.org/10.2174/1566524020666200719021653
DOI https://dx.doi.org/10.2174/1566524020666200719021653 |
Print ISSN 1566-5240 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5666 |
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