Plasma Exosomal miR-450b-5p as a Possible Biomarker and Therapeutic Target for Transient Ischaemic Attacks in Rats

J Mol Neurosci. 2019 Dec;69(4):516-526. doi: 10.1007/s12031-019-01341-9. Epub 2019 Jul 31.

Abstract

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.

Keywords: Biomarker; Droplet digital PCR; Plasma/cerebrospinal fluid exosomes; Rno-miR-450b-5p; Transient ischaemic attack.

MeSH terms

  • Animals
  • Biomarkers / blood
  • Biomarkers / cerebrospinal fluid
  • Exosomes / genetics
  • Exosomes / metabolism
  • Infarction, Middle Cerebral Artery / blood*
  • Infarction, Middle Cerebral Artery / cerebrospinal fluid
  • Infarction, Middle Cerebral Artery / diagnosis
  • MicroRNAs / blood*
  • MicroRNAs / cerebrospinal fluid
  • MicroRNAs / genetics
  • Rats

Substances

  • Biomarkers
  • MIRN450 microRNA, rat
  • MicroRNAs