The Circulating Exosomal Long RNAs Expression Reveals T Cell Mediated Inflammatory Response and The Potential Clinical Value Analysis in Ischemic Stroke

This was a case-control study conducted from August 2020 to January 2021, in which 9 ischemic stroke and 2 healthy individuals (healthy control group) attending the Guangdong Provincial People's Hospital were recruited for plasma exoLRs sequencing. The other 10 normal samples from a previous study published in the Gene Expression Omnibu (GEO) database (Accession No. GSE159657) were used as part of healthy control group to analyzed the expression profile together. 30 ischemic stroke patients and 30 healthy individuals were also recruited for validation of the expression of exoLRs. Ischemic stroke diagnosis was performed according to World Health Organization guidelines and validated using computed tomography or magnetic resonance imaging. The ischemic stroke must have lasted not more 48 hours prior to recruitment. The participants ranged between 18–75 years old and were from either gender. To be included in this study, healthy individuals had to have displayed normal renal and liver function, and with no history of smoking, malignancy, recent cardiovascular or cerebrovascular events, rheumatologic disorders, chronic heart failure, diabetes, acute or chronic infectious disease, aortic dissection, pulmonary embolism, myocarditis, pericarditis or congenital heart disease.Blood samples were collected immediately at admission before thrombolytic therapy. For each patient, 2 ml of venous blood was collected in ethylene diamine tetraacetic acid routine blood tubes. In order to extract plasma, the blood was centrifuged at 3000×g for 10 min at 4 °C and the supernatant was carefully transferred to a new tube for repeated centrifugation. Then the plasma was stored in cryogenic vials at −80 °C till further use. Total RNA in exosomal vesicles was extracted using the exoRNeasy Serum/Plasma kit (Qiagen), following the manufacturer’s protocol. The RNA-seq libraries were constructed using SMART technology (Clontech). RNA sequencing was performed by the illumine Nova-Seq 6000 System.