Clinical Medical College, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Department of Fundamental Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Int Urol Nephrol. 2018 May;50(5):973-982. doi: 10.1007/s11255-017-1779-4. Epub 2018 Jan 12.
To assess a new and highly specific, but low-cost, easily performed and suitable for large-scale applications method for renal fibrosis (RF) diagnostics.
Thirty-five RF and twenty non-RF patients were enrolled in the study. An appropriate polyethylene glycol (PEG) was used to isolate urinary exosomes. The efficiency of isolation process was evaluated by the morphology and size observation, as well as the detection of specific markers (CD63, CD9). The expression level of exosomal miR-29c, miR-21 and the endogenous control snRNA-U6 were detected by qRT-PCR. The diagnostic potency of urinary exosomal miR-29c and miR-21 was estimated by the ROC method. Spearman's rank-order correlations analysis was used to assess the correlation between the miRNAs and clinical parameters, including pathological index.
PEG-based method for isolation urinary exosome was effective and could be completed with a relatively low-speed centrifugal machine. Exosomal miR-29c and miR-21 were detected in all samples. The analysis of miRNAs in urinary exosomes revealed significant dys-regulation of miR-29c and miR-21 associated with RF. Exosomal miR-29c and miR-21 could predict degree of RF with AUC of 0.8333 and 0.7639 (P < 0.05). Correlation analysis showed that the level of miR-29c had a significant negative relationship with eGFR and the interstitial relative area.
The PEG-based method for isolation urinary exosome is an inexpensive and easily performed approach. The application for cargo miRNA analysis is feasible. Urinary exosomal miR-29c may present a promising diagnostic approach.
评估一种新的、高度特异的、但成本低、易于操作且适用于大规模应用的肾纤维化（RF）诊断方法。
本研究纳入 35 例 RF 患者和 20 例非 RF 患者。采用合适的聚乙二醇（PEG）分离尿液外泌体。通过形态和大小观察，以及特异性标志物（CD63、CD9）检测评估分离过程的效率。采用 qRT-PCR 检测外泌体 miR-29c、miR-21 和内参 snRNA-U6 的表达水平。采用 ROC 法评估尿源性外泌体 miR-29c 和 miR-21 的诊断效能。采用 Spearman 秩相关分析评估 miRNA 与包括病理指标在内的临床参数的相关性。
基于 PEG 的方法有效分离尿液外泌体，可在相对低速的离心机上完成。所有样本均检测到外泌体 miR-29c 和 miR-21。尿外泌体 miRNA 分析显示，miR-29c 和 miR-21 存在明显失调，与 RF 相关。外泌体 miR-29c 和 miR-21 可预测 RF 程度，AUC 分别为 0.8333 和 0.7639（P<0.05）。相关性分析显示，miR-29c 水平与 eGFR 和间质相对面积呈显著负相关。
基于 PEG 的方法分离尿液外泌体是一种廉价且易于操作的方法。其用于 cargo miRNA 分析是可行的。尿外泌体 miR-29c 可能是一种很有前途的诊断方法。
