Neuroglobin positively regulates YTHDF3 to restore mitophagy and suppress ferroptosis in sevoflurane-induced developmental neurotoxicity.

Sevoflurane-induced developmental neurotoxicity is characterized by impaired mitophagy and excessive ferroptosis. Neuroglobin (NGB) exerts broad neuroprotective effects, yet its exact epitranscriptomic regulatory mechanism against sevoflurane injury remains elusive. Two complementary experimental systems with identical total sevoflurane exposure length were adopted for in vitro mechanistic exploration and in vivo physiological verification, respectively. HT22 hippocampal neurons underwent uninterrupted 6 h sevoflurane incubation, while neonatal rats received 2 h intermittent inhalation each day for three consecutive days. We constructed lentiviral systems to overexpress or knockdown NGB/YTHDF3, then detected cell viability, mitochondrial function, reactive oxygen species (ROS), iron levels and pathway biomarkers. Co-immunoprecipitation verified endogenous protein interaction between NGB and m6A reader YTHDF3, and in vivo experiments further detected hippocampal mitophagy, ferroptosis indicators and mitochondrial ultrastructure. In vitro assays demonstrated that NGB alleviates sevoflurane-triggered mitochondrial damage and neuronal death via two separate cascades: NGB restores mitophagy and suppresses ferroptosis by interacting with and positively regulating the protein expression of YTHDF3, whereas its intrinsic antioxidant activity improves cell viability and eliminates ROS in a YTHDF3-independent manner. Collectively, this study describes a NGB-YTHDF3 epitranscriptomic regulatory mechanism that mitigates sevoflurane-triggered developmental neurotoxicity and offers therapeutic insights for pediatric brain protection.