Department of Neurology, The First Affiliated Hospital, Guangxi Medical University, Nanning, 530021, China.
J Mol Neurosci. 2019 Dec;69(4):516-526. doi: 10.1007/s12031-019-01341-9. Epub 2019 Jul 31.
Transient ischaemic attack (TIA) and cerebral infarction are difficult to identify within the thrombolytic time window. Blood markers are efficient, economical and noninvasive and can be beneficial in the diagnosis of many diseases. Plasma exosomal biomarkers are rarely reported in TIA. Exosomal microRNAs (miRNAs) were extracted from plasma and cerebrospinal fluid after middle cerebral artery occlusion (MCAo) in rats (0 min, 5 min, 10 min, 2 h). Deep sequencing was used to detect exosomal miRNAs in rat plasma and confirm significant differentially expressed miRNAs. Polymerase chain reaction (PCR) was used to detect the differentially expressed miRNAs in plasma and cerebrospinal fluid. Exosomal miRNAs with the same expression trends in plasma and cerebrospinal fluid were selected, and bioinformatics analysis was then carried out. Finally, the area under the curve (AUC) of the receiver operating characteristic (ROC) curve was determined to assess the diagnostic accuracy of miRNAs for TIA in rats. First, high-throughput sequencing was used to detect the expression level of plasma exosome miRNA, and rno-miR-450b-5p with a decreasing expression level was screened. Second, the expression levels of exosomal miRNAs were verified in cerebrospinal fluid and plasma samples by PCR, and the results indicated that exosomal rno-miR-450b-5p was similarly expressed in cerebrospinal fluid and plasma. ROC analysis showed high AUC values for rno-miR-450b-5p (0.880) in the 10 min ischaemia rats compared with the control rats. Finally, bioinformatic analysis indicated that exosomal rno-miR-450b-5p may be involved in cerebral ischaemia. Plasma exosomal rno-miR-450b-5p has a high diagnostic value and may become a therapeutic target for rat TIA.
短暂性脑缺血发作（TIA）和脑梗死在溶栓时间窗内难以识别。血液标志物高效、经济、无创，可有助于多种疾病的诊断。血浆外泌体生物标志物在 TIA 中很少报道。在大鼠大脑中动脉闭塞（MCAo）后（0 分钟、5 分钟、10 分钟、2 小时）提取血浆和脑脊液中的外泌体 microRNAs（miRNAs）。使用深度测序检测大鼠血浆中外泌体 miRNAs，并确认显著差异表达的 miRNAs。聚合酶链反应（PCR）用于检测血浆和脑脊液中差异表达的 miRNAs。选择血浆和脑脊液中表达趋势相同的外泌体 miRNAs，然后进行生物信息学分析。最后，通过接受者操作特征（ROC）曲线下面积（AUC）评估 miRNA 对大鼠 TIA 的诊断准确性。首先，使用高通量测序检测血浆外泌体 miRNA 的表达水平，筛选表达水平降低的 rno-miR-450b-5p。其次，通过 PCR 验证脑脊液和血浆样本中外泌体 miRNAs 的表达水平，结果表明 rno-miR-450b-5p 在脑脊液和血浆中表达相似。ROC 分析显示，与对照组相比，10 分钟缺血大鼠 rno-miR-450b-5p 的 AUC 值较高（0.880）。最后，生物信息学分析表明，外泌体 rno-miR-450b-5p 可能参与脑缺血。血浆外泌体 rno-miR-450b-5p 具有较高的诊断价值，可能成为大鼠 TIA 的治疗靶点。
