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心房颤动患者外周全血、无细胞血浆和外泌体包裹的线粒体 DNA 拷贝数的定量分析。

Quantification of peripheral whole blood, cell-free plasma and exosome encapsulated mitochondrial DNA copy numbers in patients with atrial fibrillation.

机构信息

Department of Human Genetics, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.

Institute of Cardiology, Faculty of Medicine, University of Debrecen, Debrecen, Hungary.

出版信息

J Biotechnol. 2019 Jun 20;299:66-71. doi: 10.1016/j.jbiotec.2019.04.018. Epub 2019 May 4.

Abstract

Mitochondrial DNA (mtDNA) copy number changes have been associated with various diseases. Several studies showed that mtDNA content in peripheral blood was associated with oxidative stress and cardiovascular disease. Atrial fibrillation (AF) is one of the severe cardiovascular diseases. We aimed to determine the mtDNA copy numbers in peripheral blood, in cell-free plasma and in exosomes of AF patients and healthy controls. Peripheral blood was drawn from 60 AF patients and 72 healthy controls. DNA was isolated from EDTA blood and plasma. Exosomes were isolated from cell-free plasma and then exosome encapsulated DNA (exoDNA) was extracted. Quantitative-real-time PCR was performed with Human Mitochondrial DNA (mtDNA) Monitoring Primer Set. Statistical analysis of the data was performed. We found statistically significant difference in mtDNA copy numbers in DNA isolated from peripheral whole blood, cell-free plasma and exosome samples of controls' (44.4 ± 18.0, 27.2 ± 30.1, 11.5 ± 8.7), and patients' group (43.4 ± 13.6, 26.2 ± 26.4, 14.5 ± 12.3). However there was no significant difference in mtDNA copy number between the two study groups either in peripheral blood, in cell-free plasma and in exosomes, and even in different sexes and ages. We found the highest copy number of mtDNA in peripheral blood, followed by plasma and exosomes. We did not find differences between patients and controls, neither age nor gender had effect on the mtDNA copy number. According to our results the mtDNA copy numbers did not differ in AF patients.

摘要

线粒体 DNA(mtDNA)拷贝数的变化与各种疾病有关。一些研究表明,外周血中的 mtDNA 含量与氧化应激和心血管疾病有关。心房颤动(AF)是一种严重的心血管疾病。我们旨在确定 AF 患者和健康对照者外周血、无细胞血浆和外泌体中的 mtDNA 拷贝数。从 60 例 AF 患者和 72 例健康对照者中抽取外周血。从 EDTA 血液和血浆中分离 DNA。从无细胞血浆中分离外泌体,然后提取外泌体包裹的 DNA(exoDNA)。采用人线粒体 DNA(mtDNA)监测引物组进行定量实时 PCR。对数据进行统计分析。我们发现对照组外周全血、无细胞血浆和外泌体样本中 mtDNA 拷贝数存在统计学差异(44.4±18.0、27.2±30.1、11.5±8.7),而患者组中 mtDNA 拷贝数无统计学差异(43.4±13.6、26.2±26.4、14.5±12.3)。然而,无论是在外周血、无细胞血浆还是在外泌体中,两组之间的 mtDNA 拷贝数均无显著差异,甚至在不同性别和年龄的人群中也是如此。我们发现外周血中 mtDNA 的拷贝数最高,其次是血浆和外泌体。我们未发现患者和对照组之间存在差异,年龄和性别对 mtDNA 拷贝数均无影响。根据我们的结果,AF 患者的 mtDNA 拷贝数没有差异。

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