Wang Wei, Li Dong-Bin, Li Ru-Ying, Zhou Xia, Yu Dong-Ju, Lan Xiao-Yan, Li Jin-Pin, Liu Jing-Li
Department of Neurology, The First Affiliated Hospital, Guangxi Medical University, Nanning, China.
Department of Neurology, The Third People's Hospital of Nanning, Nanning, China.
Cerebrovasc Dis. 2018;45(5-6):204-212. doi: 10.1159/000488365. Epub 2018 Apr 6.
Early and accurate diagnosis of ischaemic stroke (IS) requires the use of an optimized biomarker. Exosomal microRNAs have the potential to serve as biomarkers owing to their stability and specificity. We investigated the expression levels of plasma-derived exosomal microRNA-21-5p and microRNA-30a-5p in the different phases of IS.
One hundred forty-three patients with IS and 24 non-stroke controls were enrolled. The patients were divided into the following 5 groups: 1 group for the hyperacute phase IS (HIS, within 6 h); two for the acute phase IS (AIS, including days 1-3 and days 4-7); one for the subacute phase IS (SIS, days 8-14); and one for the recovery phase IS (RIS, days >14). Plasma exosomes were isolated using a QIAGEN exoRNeasy kit and examined by transmission electron -microscopy, nanoparticle tracking, and flow cytometry. The expression levels of miRNA-21-5p and miRNA-30a-5p were detected by quantitative real-time polymerase chain reaction.
The plasma exosomal miR-21-5p levels in SIS and RIS were significantly higher than that in controls (p < 0.05 and p < 0.01 respectively). The levels of miR-30a-5p in HIS were significantly higher (p < 0.05) and in AIS (days 1-3) were lower than that in controls (p < 0.05). In AIS (days 1-3), both miRNAs were decreased compared with the HIS group (p = 0.053 and 0.001, respectively). The area under the curve (AUC) of the miR-21-5p was 0.714 for SIS (95% CI 0.570-0.859, p = 0.007), 0.734 for RIS (95% CI 0.596-0.871, p = 0.003); the AUC of the miR-30a-5p was 0.826 for HIS (95% CI 0.665-0.988, p = 0.001), 0.438 for AIS (days 1-3; 95% CI 0.240-0.635, p = 0.516).
The plasma-derived exosomal miR-21-5p and miRNA-30a-5p in combination are promising biomarkers for diagnosing IS and distinguishing among HIS, SIS, and RIS, especially miRNA-30a-5p for the diagnosis of the HIS phase. Our results provide a new reference for clinicians to apply in early-stage diagnosis and identifies the possible value of biomarkers for IS thrombolysis therapy.
缺血性卒中（IS）的早期准确诊断需要使用优化的生物标志物。外泌体微小RNA因其稳定性和特异性有潜力作为生物标志物。我们研究了血浆来源的外泌体微小RNA-21-5p和微小RNA-30a-5p在IS不同阶段的表达水平。
纳入143例IS患者和24例非卒中对照。患者分为以下5组：1组为超急性期IS（HIS，6小时内）；2组为急性期IS（AIS，包括第1 - 3天和第4 - 7天）；1组为亚急性期IS（SIS，第8 - 14天）；1组为恢复期IS（RIS，>14天）。使用QIAGEN外泌体RNA提取试剂盒分离血浆外泌体，并通过透射电子显微镜、纳米颗粒追踪和流式细胞术进行检测。通过定量实时聚合酶链反应检测miRNA-21-5p和miRNA-30a-5p的表达水平。
SIS和RIS组血浆外泌体miR-21-5p水平显著高于对照组（分别为p < 0.05和p < 0.01）。HIS组miR-30a-5p水平显著升高（p < 0.05），AIS（第1 - 3天）组低于对照组（p < 0.05）。在AIS（第1 - 3天），与HIS组相比，两种微小RNA均降低（分别为p = 0.053和0.001）。miR-21-5p在SIS组的曲线下面积（AUC）为0.714（95%CI 0.570 - 0.859，p = 0.007），在RIS组为0.734（95%CI 0.596 - 0.871，p = 0.003）；miR-30a-5p在HIS组的AUC为0.826（95%CI 0.665 - 0.988，p = 0.001），在AIS（第1 - 3天）组为0.438（95%CI 0.240 - 0.635，p = 0.516）。
血浆来源的外泌体miR-21-5p和miRNA-30a-5p联合是诊断IS以及区分HIS、SIS和RIS的有前景的生物标志物，尤其是miRNA-30a-5p用于HIS期的诊断。我们的结果为临床医生进行早期诊断提供了新的参考，并确定了生物标志物在IS溶栓治疗中的可能价值。
