Department of Genome Medicine and Science, Gachon University College of Medicine, Incheon, Republic of Korea; Gachon Institute of Genome Medicine and Science, Gachon University Gil Medical Center, Incheon, Republic of Korea.
Department of Internal Medicine, Gachon University Gil Medical Center, Incheon, Republic of Korea; Department of Internal Medicine, Gachon University College of Medicine, Incheon, Republic of Korea.
Diabetes Res Clin Pract. 2020 Feb;160:108010. doi: 10.1016/j.diabres.2020.108010. Epub 2020 Jan 24.
MicroRNAs (miRNAs) that circulate in biological fluids are frequently enclosed in extracellular vesicles (EVs). However, urinary EVs and their cargo miRNAs have not been systematically studied according to their EV isolation methods.
In type 2 diabetes mellitus persons with diabetic nephropathy (n = 4), we compared miRNA species in urine EVs prepared by ultracentrifugation (UC), qEV original size exclusion column (qEV), ExoQuick-TC Plus (ExoQuick), and ultrafiltration using Amicon Ultra centrifugal filter devices (Amicons) 10 K and 100 K. EV miRNAs were profiled by next-generation sequencing (NGS). Additionally, we evaluated the correlations of EV miRNA expression between the urine and serum samples isolated by UC.
From each of 100 ml of urine, the UC method yielded the highest number of EV miRNA species (233 ± 37.3), with the ExoQuick yielded the lowest (103 ± 17.4). Urine EV miRNA profiles were highly correlated between UC, qEV, ExoQuick and Amicon 10 K methods. EV miRNA profiles between the urine and serum samples showed variable correlations between the patients (paired sample number = 3, r = 0.39-0.72).
UC, qEV, ExoQuick, and Amicon 10 K are acceptable for urinary EV isolation to profile miRNAs. Urine- and serum-derived EV miRNA profiles have variable correlations depending on specific patients.
存在于生物体液中的 microRNAs（miRNAs）经常被包裹在细胞外囊泡（EVs）中。然而，根据其 EV 分离方法，尚未系统地研究尿液 EVs 及其携带的 cargo miRNAs。
在 2 型糖尿病合并糖尿病肾病患者（n=4）中，我们比较了通过超速离心（UC）、qEV 原始大小排阻柱（qEV）、ExoQuick-TC Plus（ExoQuick）和使用 Amicon Ultra 离心超滤器（Amicon）10 K 和 100 K 分离的尿液 EVs 中 miRNA 种类。通过下一代测序（NGS）对 EV miRNAs 进行分析。此外，我们评估了 UC 分离的尿液和血清样本中 EV miRNA 表达之间的相关性。
从每份 100 ml 的尿液中，UC 方法产生的 EV miRNA 种类最多（233±37.3），而 ExoQuick 方法产生的最少（103±17.4）。UC、qEV、ExoQuick 和 Amicon 10 K 方法之间的尿液 EV miRNA 图谱高度相关。尿液和血清样本之间的 EV miRNA 图谱在患者之间显示出不同的相关性（配对样本数=3，r=0.39-0.72）。
UC、qEV、ExoQuick 和 Amicon 10 K 是用于分离尿液 EV 以分析 miRNA 的可接受方法。尿液和血清来源的 EV miRNA 图谱与特定患者的相关性不同。
