LncRNA GAS5 Silencing Attenuates Oxygen-Glucose Deprivation/Reperfusion-Induced Injury in Brain Microvascular Endothelial Cells via miR-34b-3p-Dependent Regulation of EPHA4

• Published in: Neuropsychiatric disease and treatment 2021-01, Vol.17, p.1667-1678
• Main Authors: Shen, Bin, Wang, Lan, Xu, Yuejun, Wang, Hongwei, He, Shiyi
• Format: Article
• Language: English
• Subjects: Apoptosis; Binding sites; Brain injury; Care and treatment; Cell culture; Cell receptors; Cell viability; Complications and side effects; Endocrine system; Endothelial cells; Endothelin 1; Endothelium; EphA4 protein; Flow cytometry; Genetic aspects; Glucose; Health aspects; Hypoglycemia; Hypoxia; Immunoprecipitation; Ischemia; Medical research; Medicine, Experimental; MicroRNAs; Microvasculature; miRNA; Nitric oxide; Non-coding RNA; Original Research; Oxygen; Polymerase chain reaction; Proteins; Reperfusion; Reperfusion injury; RNA; Stroke (Disease); Variance analysis
• ISSN: 1176-6328; 1178-2021; 1178-2021
• DOI: 10.2147/NDT.S302314

The aim of our study was to explore the role of long non-coding RNA (lncRNA) growth arrest-specific 5 (GAS5) in ischemic stroke using oxygen-glucose deprivation/reperfusion (OGD/R)-induced bEnd.3 cells as in vitro cell model. Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot assay were adopted to analyze RNA and protein expression. Cell viability and apoptosis were analyzed by Cell Counting Kit-8 (CCK8) assay and flow cytometry. The levels of nitric oxide (NO) and endothelin-1 (ET-1) in culture supernatant were examined by their matching commercial kits. The intermolecular target interaction was predicted by starBase software and tested by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. OGD/R-induced apoptosis and dysregulation in vascular endocrine system were largely alleviated by the knockdown of GAS5. GAS5 interacted with microRNA-34b-3p (miR-34b-3p), and GAS5 silencing protected bEnd.3 cells from OGD/R-induced injury partly through up-regulating miR-34b-3p. EPH receptor A4 (EPHA4) was a target of miR-34b-3p. GAS5 acted as the molecular sponge of miR-34b-3p to up-regulate EPHA4 in bEnd.3 cells. GAS5 interference protected against OGD/R-induced damage in bEnd.3 cells partly through down-regulating EPHA4. LncRNA GAS5 knockdown protected brain microvascular endothelial cells bEnd.3 from OGD/R-induced injury depending on the regulation of miR-34b-3p/EPHA4 axis.