Department of Blood Transfusion, Fujian Medical University Union Hospital, Gulou District, Fuzhou City, Fujian Province 350001, China.
Department of Oncology, Quanzhou First Hospital Affiliated to Fujian Medical University, Quanzhou City, Fujian Province 362000, China.
Biomed Res Int. 2019 Jun 13;2019:2045915. doi: 10.1155/2019/2045915. eCollection 2019.
To elucidate the microRNAs existent in exosomes derived from stored red blood cell (RBC) unit and their potential function.
Exosomes were isolated from the supernatant derived from stored RBC units by sequential centrifugation. Isolated exosomes were characterized by TEM (transmission electron microscopy), western blotting, and DLS (dynamic light scattering). MicroRNA (miRNA) microarray was performed to detect the expression of miRNAs in 3 exosome samples. Results revealed miRNAs that were simultaneously expressed in the 3 exosome samples and were previously reported to exist in mature RBCs. Functions and potential pathways of some detected miRNAs were illustrated by bioinformatic analysis. Validation of the top 3 abundant miRNAs was carried out by qRT-PCR (quantitative reverse transcription-polymerase chain reaction).
TEM and DLS revealed the mean size of the exosomes (RBC-derived) as 64.08 nm. These exosomes exhibited higher abundance of short RNA than the long RNA. 78 miRNAs were simultaneously detected in 3 exosome samples and mature RBCs. Several biological processes might be impacted by these miRNAs, through their target gene(s) enriched in a particular signalling pathway. The top 3 (abundant) miRNAs detected were as follows: miR-125b-5p, miR-4454, and miR-451a. qRT-PCR revealed higher abundance of miR-451a than others. Only miR-4454 and miR-451a abundance tended to increase with increasing storage time.
Exosomes derived from stored RBC units possessed multiple miRNAs and, hence, could serve various functions. The function of exosomes (RBC-derived) might be implemented partly by the predominantly enriched miR-451a.
阐明储存的红细胞（RBC）单位衍生的外泌体中存在的 microRNA 及其潜在功能。
通过连续离心从储存的 RBC 单位上清液中分离出外泌体。通过透射电子显微镜（TEM）、western blot 和动态光散射（DLS）对分离的外泌体进行表征。通过 microRNA（miRNA）微阵列检测 3 个外泌体样本中 miRNA 的表达。结果揭示了在 3 个外泌体样本中同时表达的 miRNA，这些 miRNA 先前被报道存在于成熟的 RBC 中。通过生物信息学分析说明了一些检测到的 miRNA 的功能和潜在途径。通过 qRT-PCR（定量逆转录-聚合酶链反应）对前 3 个丰富 miRNA 进行验证。
TEM 和 DLS 显示（RBC 衍生的）外泌体的平均大小为 64.08nm。这些外泌体表现出比长 RNA 更高的短 RNA 丰度。在 3 个外泌体样本和成熟的 RBC 中同时检测到 78 个 miRNA。这些 miRNA 通过其在特定信号通路中富集的靶基因，可能影响几个生物学过程。检测到的前 3 个（丰富的）miRNA 如下：miR-125b-5p、miR-4454 和 miR-451a。qRT-PCR 显示 miR-451a 的丰度高于其他 miRNA。只有 miR-4454 和 miR-451a 的丰度随着储存时间的增加而增加。
储存的 RBC 单位衍生的外泌体含有多种 miRNA，因此可以发挥多种功能。外泌体（RBC 衍生的）的功能可能部分通过主要富集的 miR-451a 来实现。
