Abstract

Pyroptosis is a pathological process contributing to cerebral ischemia/reperfusion (I/R) injury. YTHDF1 participates in various pathological mechanisms including pyroptosis; however, its regulatory role in pyroptosis during cerebral I/R injury remains poorly understood. This study aimed to investigate the functional role and underlying molecular mechanisms of YTHDF1 in cerebral I/R injury with a focus on neuronal pyroptosis. I/R-induced rat models and oxygen-glucose deprivation and reoxygenation (OGD/R)-induced neuronal cell models were established. Infarct areas were visualized using TTC staining. Cellular viability, oxidative stress levels, and pyroptotic activity were evaluated through gain- and loss-of-function experiments. The regulatory relationship between YTHDF1 and NEK7 was analyzed using RNA immunoprecipitation, dual-luciferase reporter assays, and RNA stability detection. Our results demonstrated that YTHDF1 expression was significantly upregulated in both I/R rat models and OGD/R-treated cells. YTHDF1 knockdown suppressed pyroptotic processes while enhancing antioxidant capacity. Furthermore, YTHDF1 physically interacted with NEK7, specifically binding to the m6A modification site within Nek7 mRNA to maintain its RNA stability. NEK7 overexpression effectively counteracted the anti-pyroptotic effects induced by YTHDF1 silencing. In vivo experiments confirmed that YTHDF1 inhibition reduced infarct volume and attenuated pyroptosis in cerebral I/R injury via decreasing NEK7 expression. These findings demonstrate that YTHDF1 promotes neuronal pyroptosis by stabilizing NEK7, thereby exacerbating cerebral I/R injury, suggesting its potential as a therapeutic target for ischemic stroke treatment.