Intranasal Administrations of AP39-Loaded Liposomes Selectively Deliver H2S to Neuronal Mitochondria to Protect Neonatal Hypoxia–Ischemia by Targeting ERK1/2 and Caspase-1

Mitochondrial dysfunction contributes to the pathology of hypoxia–ischemia (HI) brain damage by aberrant production of ROS. Hydrogen sulfide (H2S) has been demonstrated to exert neuroprotective effects through antioxidant mechanisms. However, the diffusion of H2S in vivo is not specifically targeted and may even be systemically toxic. In this study, based on mitochondria-targeted H2S donor AP39, we fabricated liposomes encapsulating AP39 (AP39@Lip) via intranasal delivery to improve functional recovery after HI brain injury. This study presents that intranasal administration of AP39@Lip was capable of attenuating acute brain injury by inhibiting mitochondrial dysfunction, apoptosis, neuroinflammation, and ROS production in the lesional cortex 3 days after HI brain injury. Similarly, AP39@Lip was observed to restore both short- and long-term function following HI injury without obvious toxicity. Mechanistically, the therapeutic effects of AP39@Lip mainly relied on its colocalization with neuronal mitochondria 24 h after administration and reversed H2S levels in the lesional cortex. Moreover, molecular docking and cellular thermal shift assay suggest that AP39 inhibited the activation of ERK1/2 and caspase-1 by directly binding to ERK1/2 or caspase-1. These results indicate that intranasal administration of AP39@Lip selectively delivered H2S to neuronal mitochondria and mitigated mitochondrial damage following HI insult by targeting ERK1/2 and caspase-1.