Abstract
Cerebral ischemic stroke is a leading cause of neurological disability worldwide. Previous study reported that long noncoding RNA (lncRNA) growth arrest-specific transcript 5 (GAS5) was highly expressed in ischemic stroke. However, the mechanism underlying GAS5 in an inflammatory injury during an ischemic stroke remains poorly understood. An in vivo mouse model of middle cerebral artery occlusion (MCAO) and an in vitro cell model of oxygen-glucose deprivation (OGD) were established to induce cerebral ischemic stroke condition. The expressions of GAS5, microRNA-9 (miR-9) and forkhead box O3 (FOXO3) were measured by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot analysis, respectively. The neurological injury in vivo was investigated by neurological score and TTC staining. Cell apoptosis and inflammatory injury were analyzed by western blot, flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively. The interaction between miR-9 and GAS5 or FOXO3 was explored by luciferase activity, RNA pull-down and RNA immunoprecipitation (RIP) assays. GAS5 expression was enhanced in the cerebral ischemic stroke model. Knockdown of GAS5 attenuated the cerebral infarct, neurological injury, apoptosis and inflammatory injury in the mouse MCAO model. miR-9 was bound to GAS5 and its overexpression inhibited cell apoptosis and inflammatory response in OGD-treated bEnd.3 cells, which was attenuated by GAS5. FOXO3 was a target of miR-9 and its restoration reversed the miR-9-mediated suppression of apoptosis and inflammation. Moreover, GAS5 promoted FOXO3 expression by competitively sponging miR-9. GAS5 knockdown alleviated neuronal cell injury by regulating miR-9/FOXO3, providing a new theoretical foundation for cerebral ischemic stroke.