Liproxstatin-1 ameliorates cerebral ischemia-reperfusion injury through inhibiting ferroptosis.

OBJECTIVE This study intends to investigate the protective impact of liproxstatin-1 against cerebral ischemia-reperfusion injury (CIRI) in rats and the corresponding underlying mechanism.

METHODS CIRI rat models were constructed. Pathological changes in tissues were assessed at multiple levels, including infarct area, neuronal activity, and iron content through 2,3,5-triphenyl tetrazolium chloride (TTC) staining, Nissl staining, and Prussian blue (PB) staining. The expression of oxidative damage factors and key ferroptosis-related genes was assessed by enzyme-linked immunosorbent assay (ELISA), quantitative real-time PCR (qPCR), or western blot (WB). Rat brain tissues were subjected to bulk RNA sequencing (bulk RNA-seq) analysis. Finally, in vitro oxygen-glucose deprivation/reoxygenation (OGD/R) models were established, and the inhibitory effect of liproxstatin-1 on ferroptosis was identified by measuring cell viability, Fe2+ levels, and lipid peroxidation.

RESULTS In vivo experiments demonstrated that liproxstatin-1 markedly improved neurological function and neuronal pathological damage in CIRI rats, while reducing iron content and oxidative damage. Bulk RNA-seq analysis revealed that differentially expressed genes (DEGs) were mainly enriched in the IL-17 signaling pathway, Ether lipid metabolism, TNF signaling pathway, and ferroptosis. Consistently, qPCR and WB results showed that liproxstatin-1 treatment increased the expression of FTH1 and GPX4, while decreasing the expression of NOX1, ACSL4, COX2, and TFR1 in rat brain tissues. In vitro experiments further demonstrated that liproxstatin-1 significantly enhanced cell viability and reduced Fe2+ and reactive oxygen species (ROS) levels.

CONCLUSION Liproxstatin-1 inhibits the process of ferroptosis and alleviates CIRI in rats.