YZG-331 alleviates sleep disorders in pain mice by inhibiting N-methyl-D-aspartate-mediated neurotoxicity

Ran Guangyuan; Liu Yajing; Fu Zhouyang; Wu Hao; Sun Haiyan; He Mingwei

doi:10.3760/j.issn.1673-4777.2025.11.009

Abstract：Objective To explore the effect of YZG-331 on pain threshold and sleep quality in a mouse model of pain and its potential mechanism.Methods Thirty-six male specific pathogen-free 8-week-old C57BL/6J mice were selected.They were randomly divided into sham operation group,operation group and observation group,with 12 mice in each group.In the sham operation group,the skin was only cut and sutured after disinfection with iodophor.In the operation group and the observation group,L4/L5 dorsal root ganglion was cut and scraped under microscopic conditions to establish a pain mouse model.The mice in the observation group were intraperitoneally injected with YZG-331 at a concentration of 10 mg/kg on day 1,2,and 3 after operation,and the solvent was sodi-um carboxymethyl cellulose.The mice in the operation group and the sham operation group were given the same vol-ume of sodium carboxymethyl cellulose.Patch clamp was used to detect neuronal electrical signals,and proteomics analysis,molecular docking and kinetic simulation electrophysiological experiments were performed to evaluate the activity of N-methyl-D-aspartate(NMDA)receptor and the mechanism of YZG-331 on it.Results YZG-331 significantly increased the prickling and thermal pain thresholds,improved sleep quality,and inhibited pain-induced abnormalities in brain electrical activity in mice.Proteomics sequencing,molecular docking and electrophysiological experiments showed that YZG-331 stably bound to NMDA receptors.The docking score of YZG-331 with NMDA was-4.742,and the result of generalized Born surface area analysis was-33.08 kcal/mol.It penetrated deep into the receptor active pocket and formed a variety of interactions with key amino acid residues to inhibit NMDA receptor-mediated neurotoxicity and regulate cellular excitability balance.Conclusion The present findings provide strong experimental evidence for YZG-331 as a potential therapeutic agent for pain and sleep disorders.