Therapeutic hypothermia (TH) is the only intervention approved for the treatment of neonatal hypoxic-ischaemic encephalopathy (HIE), but its treatment window is narrow (within 6 h after birth), and its efficacy is not ideal. Thus, alternative treatments are urgently needed. Our previous studies showed that genistein-3′-sodium sulfonate (GSS), a derivative of genistein (Gen), has a strong neuroprotective effect in rats with ischaemic stroke, but its role in HIE is unclear. A hypoxia–ischaemia (HI) brain injury model was established in neonatal male Sprague‒Dawley (SD) rats. Twenty-four hours after reperfusion, rats treated with GSS were assessed for cerebral infarction, neurological function, and neuronal damage. RNA-Seq and bioinformatics analysis were used to explore differentially expressed genes (DEGs) and regulated signalling pathways, which were subsequently validated by Western blotting and immunofluorescence. In this study, we found that GSS not only significantly reduced the size of brain infarcts and alleviated nerve damage in rats with HIE but also inhibited neuronal loss and degeneration in neonatal rats with HIE. A total of 2170 DEGs, of which 1102 were upregulated and 1068 were downregulated, were identified in the GSS group compared with the HI group. In an analysis based on Kyoto Encyclopedia of Genes and Genomes (KEGG) categories, the downregulated DEGs were significantly enriched in the pathways “Phagosome”, “NF-κB signalling”, and “Complement and coagulation cascades”, amongst others. Meanwhile, the upregulated DEGs were significantly enriched in the pathways “Neurodegeneration”, “Glutamatergic synapse”, and “Calcium signalling pathway”, amongst others. These results indicate that GSS intervenes in the process of HIE-induced brain injury by participating in multiple pathways, which suggests potential candidate drugs for the treatment of HIE. Graphical Abstract

Background Currently, therapeutic hypothermia (TH) is the only intervention approved for the treatment of neonatal hypoxic-ischaemic encephalopathy (HIE), but the treatment window for TH is narrow (within 6 h after birth), and its efficacy is not ideal. Thus, alternative treatments are urgently needed., Our previous studies showed that Genistein-3′-sodium sulfonate (GSS), a derivative of genistein (Gen), has a strong neuroprotective effect in ischemia stroke rats, but its role in HIE is unclear. Methods The HI brain injury model was established in male Sprague-Dawley (SD) neonatal rats. Following treatment with GSS, cerebral infarction, neurological function, neuron damaged were assessed 24 h after reperfusion. RNA-Seq and bioinformatics analysis were used to explore differential expression genes (DEGs) and signals, which were validated by Western blotting subsequently. Results In this study, we found that GSS not only significantly reduced brain infarct size of rats with HIE and alleviated nerve damage, but also inhibited neuronal loss and degeneration in neonatal rats with HIE. A total of 2170 DEGs, of which 1102 were upregulated and 1068 were downregulated, were identified in the GSS group compared with the HI group. Downregulated DEGs significantly enriched in the Phagosome, NF-κB signalling pathway, Complement and coagulation cascades, and so on. Upregulated DEGs were significantly enriched in the Pathways of neurodegeneration, Glutamatergic synapse, Calcium signalling pathway, and so on. Conclusion These results indicate that GSS intervenes the process of HIE-induced brain injury by participating in multiple pathways, which provided drug candidates for the treatment of HIE.