Esculetin inhibits ferroptosis after ICH by promoting NUDT1-mediated m7G methylation modification of GPX4

Intracerebral hemorrhage (ICH) is the most morbid of all stroke types with a high mortality and significant disability burden. Inhibiting ferroptosis after ICH will effectively treat ICH and improve prognosis. Esculetin is the main active ingredient of the Chinese medicine Cortex Fraxini, which has neuroprotective effects. 7-methylguanosine (m7G) methylation is a common RNA modification that regulates mRNA stability and translation, playing an important role in neural development and function, as well as in ferroptosis-related pathways. In this study, we used hemin-induced PC12 cells to investigate the impact of esculetin on ferroptosis and the involvement of m7G modification. Cell viability was measured by cell counting kit-8. Cell death was evaluated by measuring the levels of LDH release and PI-positive cells. Ferroptosis was assessed by measuring the levels of Fe²⁺, glutathione, lipid reactive oxygen species, and malondialdehyde. The underlying mechanism was investigated by quantitative real-time PCR, methylated RNA immunoprecipitation (MeRIP), and RIP. Results suggested that esculetin promoted cell viability, inhibited ferroptosis, and increased NUDT1-mediated m7G levels in hemin-induced PC12 cells. Besides, NUDT1 interacted with GPX4 and enhanced the mRNA stability of GPX4. Silencing of GPX4 abolished the inhibitory influences of NUDT1 overexpressing on ferroptosis in hemin-induced PC12 cells. The ICH mouse model revealed that esculetin treatment effectively reduced neurological deficits and inhibited ferroptosis. In conclusion, esculetin treatment inhibits ICH-induced ferroptosis by promoting NUDT1-mediated m7G methylation modification and enhancing GPX4 stability. This study contributes to understanding the mechanisms by which esculetin may mitigate ICH damage and may provide a new potential therapeutic target for ICH.