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毛细管电泳技术在 2 型糖尿病患者血液中糖基化分析的初步研究。

-Glycosylation Profiling of Human Blood in Type 2 Diabetes by Capillary Electrophoresis: A Preliminary Study.

机构信息

Translational Glycomics Research Group, Research Institute of Biomolecular and Chemical Engineering, University of Pannonia, 8200 Veszprem, Hungary.

Horvath Csaba Memorial Laboratory of Bioseparation Sciences, Research Center for Molecular Medicine, Faculty of Medicine, Doctoral School of Molecular Medicine, University of Debrecen, 4032 Debrecen, Hungary.

出版信息

Molecules. 2021 Oct 22;26(21):6399. doi: 10.3390/molecules26216399.

Abstract

Currently, diagnosing type 2 diabetes (T2D) is a great challenge. Thus, there is a need to find rapid, simple, and reliable analytical methods that can detect the disease at an early stage. The aim of this work was to shed light on the importance of sample collection options, sample preparation conditions, and the applied capillary electrophoresis bioanalytical technique, for a high-resolution determination of the -glycan profile in human blood samples of patients with type 2 diabetes (T2D). To achieve the profile information of these complex oligosaccharides, linked by asparagine to hIgG in the blood, the glycoproteins of the samples needed to be cleaved, labelled, and purified with sufficient yield and selectivity. The resulting samples were analyzed by capillary electrophoresis, with laser-induced fluorescence detection. After separation parameter optimization, the capillary electrophoresis technique was implemented for efficient -glycan profiling of whole blood samples from the diabetic patients. Our results revealed that there were subtle differences between the -glycan profiles of the diabetic and control samples; in particular, two -glycan structures were identified as potential glycobiomarkers that could reveal significant changes between the untreated/treated type 2 diabetic and control samples. By analyzing the resulting oligosaccharide profiles, clinically relevant information was obtained, revealing the differences between the untreated and HMG-CoA reductase-inhibitor-treated diabetic patients on changes in the -glycan profile in the blood. In addition, the information from specific IgG -glycosylation profiles in T2D could shed light on underlying inflammatory pathophysiological processes and lead to drug targets.

摘要

目前,诊断 2 型糖尿病(T2D)是一个巨大的挑战。因此,需要寻找快速、简单和可靠的分析方法,以便在早期阶段检测出这种疾病。本工作旨在阐明样品采集选择、样品制备条件以及所应用的毛细管电泳生物分析技术对于高分辨率测定 2 型糖尿病患者人血样中-β-聚糖谱的重要性。为了获得这些通过天冬酰胺与 hIgG 在血液中连接的复杂寡糖的谱信息,需要对样品中的糖蛋白进行切割、标记和纯化,以获得足够的产率和选择性。用毛细管电泳,结合激光诱导荧光检测对所得样品进行分析。在分离参数优化后,将毛细管电泳技术用于从糖尿病患者的全血样品中进行有效的-β-聚糖谱分析。我们的结果表明,糖尿病和对照样品的-β-聚糖谱之间存在细微差异;特别是,有两种-β-聚糖结构被鉴定为潜在的糖生物标志物,它们可以揭示未经处理/经处理的 2 型糖尿病患者和对照样品之间的显著变化。通过分析所得的寡糖谱,获得了临床相关信息,揭示了未经处理和 HMG-CoA 还原酶抑制剂治疗的糖尿病患者在血液中-β-聚糖谱变化方面的差异。此外,T2D 中特定 IgG-糖基化谱的信息可以揭示潜在的炎症病理生理过程,并为药物靶点提供信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8191/8586923/c1f4cb037281/molecules-26-06399-g001.jpg

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