Multi-omics network pharmacology and experimental verification of erythropoietin in microglial neuroinflammation in neonatal hypoxic-ischemic brain injury
Lingzhi Wang, Mian Chen, Luying Yang, Tianyi Miao, Li Sun, Xiaoyu RuanAbstract
Objectives
This study aims to investigate the neuroprotective mechanisms of erythropoietin (EPO) in neonatal hypoxic-ischemic brain injury (HIBI), with a particular focus on its role in regulating microglia-mediated neuroinflammation.
Methods
The common targets of EPO and HIBI were screened through databases, and a PPI network was constructed. GO and KEGG pathway enrichment analysis were performed. The binding characteristics of EPO to the core target were evaluated using molecular docking and kinetic simulation. In vivo validation was performed in a neonatal rat HIBI model to detect microglial activation, the NF-κB pathway, cytokine levels, neuronal apoptosis, and motor function recovery.
Results
92 overlapping targets were identified, with AKT1, STAT3, TNF, IL-6, and NFKB1 as hub genes, mainly enriched in PI3K-AKT, NF-κB, and JAK-STAT pathways. EPO had strong binding affinity with these core targets (−7.4 ∼ −9.1 kcal/mol), and kinetic simulation confirmed that the binding was stable. In vivo experiments showed that EPO reduced Iba-1 + microglia via suppressing the NF-κB pathway, promoted their polarization to M2 phenotype, down-regulated pro-inflammatory cytokines, reduced neuronal apoptosis, and dose-dependently improved motor function.
Conclusions
EPO polarizes microglia into the M2 phenotype by regulating NF-κB signaling and reducing neuroinflammation, threrby playing a neuroprotective role.