LncRNA GAS6-AS1 promotes ischemic stroke injury through miR-370-3p/TFRC-mediated ferroptosis and disruption of Nrf2/ARE antioxidant responses

Abstract

Ischemic stroke induces severe neuronal injury after ischemia/reperfusion (I/R), with oxidative disturbance and ferroptotic death emerging as critical contributors. Although GAS6-AS1 has been associated with ischemic stroke progression, the molecular mechanisms underlying its function in neuronal injury remain to be elucidated. An in vitro ischemic model was established by exposing SH-SY5Y neuronal cells to oxygen–glucose deprivation/reperfusion (OGD/R) conditions, followed by genetic modulation of GAS6-AS1 to assess its functional role. Neuronal damage, iron accumulation, lipid oxidation, glutathione homeostasis, and ferroptosis-associated proteins were examined by biochemical assays and Western blotting. Molecular interactions among GAS6-AS1, miR-370-3p, and TFRC were validated by luciferase reporter, RNA immunoprecipitation, and qRT-PCR assays. In vivo effects were evaluated in MCAO rats receiving AAV-sh-GAS6-AS1. The clinical association of these molecules was further examined in plasma samples from 35 patients with ischemic stroke and matched controls. GAS6-AS1 levels showed consistent upregulation across OGD/R-treated neuronal cells, MCAO brain tissues, and plasma samples from patients with ischemic stroke, accompanied by reduced miR-370-3p and increased TFRC levels. GAS6-AS1 overexpression aggravated ferroptotic alterations and disrupted Nrf2/ARE-related antioxidant responses. Conversely, miR-370-3p restoration, TFRC suppression, or Fer-1 treatment attenuated ferroptosis-associated changes and partially recovered antioxidant capacity. GAS6-AS1 silencing alleviated neurological deficits, reduced infarct injury, and improved ferroptosis-related abnormalities in MCAO rats. In patients, GAS6-AS1 and TFRC levels positively correlated, whereas miR-370-3p levels inversely correlated with NIHSS scores. Our data indicate that GAS6-AS1 facilitates ischemic neuronal injury through modulation of the miR-370-3p/TFRC pathway and ferroptosis-associated responses, together with changes in Nrf2/ARE antioxidant activity. These results support the involvement of GAS6-AS1 in ischemic stroke pathology and suggest its potential value for future therapeutic exploration.