Department of Nephrology, Huzhou Central Hospital, Affiliated Central Hospital Huzhou University, Huzhou 313000, Zhejiang, China.
Genet Res (Camb). 2022 Aug 31;2022:2276175. doi: 10.1155/2022/2276175. eCollection 2022.
Ultrafiltration failure remains one of the most severe complications of long-term peritoneal dialysis (PD), which results in death. This study aimed to characterize the circulating exosomal microRNA (miRNA) profiles associated with ultrafiltration failure and explore its underlying mechanisms.
Exosomes were isolated from the peritoneal dialysis effluent (PDE) of patients with ultrafiltration failure or success using the ultracentrifugation method, and then transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and western blot were used for exosome characterization. After that, the isolated exosomes were sent for small RNA sequencing, and eight differentially expressed miRNAs (DE-miRNAs) were chosen for RT-qPCR validation.
TEM, NTA, and western blot revealed that exosomes were successfully isolated. After sequencing, 70 DE-miRNAs involved in ultrafiltration were identified, including 41 upregulated ones and 29 downregulated ones. Functional analyses revealed that these DE-miRNAs were significantly enriched in pathways of cancer, ubiquitin-mediated proteolysis, axon orientation, and the Rap1 and Ras signaling pathways. In addition, the consistency rate of RT-qPCR and sequencing results was 75%, which indicated the relatively high reliability of the sequencing data.
Our findings implied that these DE-miRNAs may be potential biomarkers of ultrafiltration failure, which would help us to discover novel therapeutic targets/pathways for ultrafiltration failure in patients with end-stage renal disease.
超滤衰竭仍然是长期腹膜透析（PD）最严重的并发症之一，可导致死亡。本研究旨在描述与超滤衰竭相关的循环外泌体 microRNA（miRNA）谱，并探讨其潜在机制。
使用超速离心法从超滤衰竭或成功患者的腹膜透析液（PDE）中分离外泌体，然后使用透射电子显微镜（TEM）、纳米颗粒跟踪分析（NTA）和 Western blot 对外泌体进行特征描述。之后，对分离出的外泌体进行小 RNA 测序，选择 8 个差异表达 miRNA（DE-miRNA）进行 RT-qPCR 验证。
TEM、NTA 和 Western blot 显示成功分离出外泌体。测序后，鉴定出 70 个与超滤相关的 DE-miRNA，包括 41 个上调的和 29 个下调的。功能分析表明，这些 DE-miRNA 显著富集在癌症、泛素介导的蛋白水解、轴突取向以及 Rap1 和 Ras 信号通路等途径中。此外，RT-qPCR 和测序结果的一致性率为 75%，这表明测序数据具有较高的可靠性。
我们的研究结果表明，这些 DE-miRNA 可能是超滤衰竭的潜在生物标志物，有助于发现终末期肾病患者超滤衰竭的新治疗靶点/途径。
