Abstract
BACKGROUND: Intracerebral hemorrhage (ICH) is a severe condition characterized by elevated mortality and disability rates. The cAMP-activated exchange protein-1 (EPAC-1) is implicated in various cytoprotective mechanisms; however, its specific role in ICH remains unclear. METHODS: A rat model of ICH was established by injecting autologous blood, while the in vitro primary neuronal model was stimulated using oxyhemoglobin (OxyHb). The construction of EPAC-1 overexpression wild-type (WT) and phosphorylated mutant plasmids (S108A or S108E), as well as lentiviruses, was performed for in vitro and in vivo studies. RESULTS: The cAMP signaling pathway was found to be significantly enriched following ICH by high-throughput sequencing analysis. Our findings showed that while EPAC-1 protein levels remained relatively unchanged after ICH, RabGEF activity was conspicuously upregulated. This was accompanied by a marked decrease in EPAC-1 phosphorylation levels. Mutations that activate EPAC-1 phosphorylation led to significant improvements in neuronal survival and behavioral outcomes after ICH. Bioinformatics analysis revealed that ferroptosis was significantly enriched after ICH and showed a positive correlation with EPAC-1 serine phosphorylation. EPAC-1 phosphorylation activating mutations inhibit neuronal ferroptosis, whereas inactivating mutations exacerbate it. CONCLUSION: The phosphorylation of EPAC-1 is essential for maintaining neuronal survival, which may be related to ferroptosis inhibition after ICH.