Affiliated Hospital of Chengdu University of T. C. M./Clinical Medical College, Chengdu University of T. C. M., Chengdu 610072, China.
Affiliated Hospital of Chengdu University of T. C. M./Clinical Medical College, Chengdu University of T. C. M., Chengdu 610072, China.
Exp Mol Pathol. 2018 Oct;105(2):223-228. doi: 10.1016/j.yexmp.2018.08.004. Epub 2018 Aug 16.
Renal interstitial fibrosis (RIF) is difficult to diagnose. This paper explored liquid biopsy markers in urinary exosomes derived from RIF patients. Urine samples from 32 patients with various degrees of RIF and 20 non-RIF patients were collected. The size and morphology of urinary exosomes isolated by polyethylene glycol were observed with electron microscopy. Protein biomakers of exosomes were analyzed by Western blot. qRT-PCR was used to detect the levels of biomarkers (miR-29c, miR-21, E-cadherin, and vimentin) of epithelial mesenchymal transition in urinary exosomes. The diagnostic value was detected with ROC curves. Results displayed successfully isolated urinary exosomes. The examined miRNAs and mRNAs were checked from all urinary exosomes samples, except for two cases of RIF which lacked E-cadherin mRNA. RNA levels were different in patients with diverse degrees of RIF. Urinary miR-29c was decreased with the progress of fibrosis. Levels of E-cadherin mRNA were first decreased and then increased. The contents of miR-21 and vimentin mRNA were also depended on the degrees of RIF. ROC curve analysis showed that the area under the curve (AUC) of miR-29c was 0.8621, statistically significant compared with the non-RIF group (P < 0.05). The miR-29c level within the urinary exosomes is a promising marker for the diagnosis of RIF.
肾间质纤维化（RIF）难以诊断。本文探讨了源自 RIF 患者尿液外泌体的液体活检标志物。收集了 32 名不同程度 RIF 患者和 20 名非 RIF 患者的尿液样本。用电子显微镜观察聚乙二醇分离的尿液外泌体的大小和形态。通过 Western blot 分析外泌体的蛋白生物标志物。用 qRT-PCR 检测尿液外泌体上皮间质转化生物标志物（miR-29c、miR-21、E-钙黏蛋白和波形蛋白）的水平。用 ROC 曲线检测诊断价值。结果成功分离出尿液外泌体。检查了所有尿液外泌体样本中的 miRNA 和 mRNA，但 RIF 的两个病例缺乏 E-钙黏蛋白 mRNA。不同程度 RIF 的患者 RNA 水平不同。尿 miR-29c 随着纤维化的进展而减少。E-钙黏蛋白 mRNA 水平先降低后升高。miR-21 和波形蛋白 mRNA 的含量也取决于 RIF 的程度。ROC 曲线分析显示，miR-29c 的曲线下面积（AUC）为 0.8621，与非 RIF 组相比具有统计学意义（P<0.05）。尿液外泌体中的 miR-29c 水平是诊断 RIF 的有前途的标志物。
