关键词:

"> 桂枝加龙骨牡蛎汤, 失眠, 神经炎症, PI3K/ AKT 信号通路, 网络药理学

Abstract:

This study aims to investigate the mechanism of Guizhi Jia Longgu Muli Decoction (GLMD)in treating insomnia by integrating network pharmacology,molecular docking,and experimental validation. Potential targets of GLMD against insomnia were screened using databases including TCMSP and BATMAN-TCM,followed by GO and KEGG enrichment analyses of core targets. Binding affinities between active constituents and targets were evaluated via molecular docking and molecular dynamics (MD) simulations. Insomnia was induced using a modified multiple platform water environment (MMPE) model,and the therapeutic effects of GLMD were validated through experimental assays,including ELISA and Western blot. Topological analysis revealed that core targets,such as interleukin-1β (IL-1β),IL-6,serine/ threonine kinase 1 (AKT1),and tumor necrosis factor ( TNF), are closely linked to the pathological mechanisms of insomnia. Key components, such as quercetin and kaempferol,exert anti-inflammatory effects by modulating signaling pathways,including TNF and phosphoinositide 3-kinase (PI3K) / AKT. Molecular docking indicated that the binding energies of active constituents to these targets were consistently below -5. 0 kcal/ mol,with binding stability further confirmed by MD simulations. In vivo experiments based on the MMPE model demonstrated that GLMD significantly attenuated anxiety-like behaviors and suppressed the elevation of serum pro-inflammatory cytokines (IL-6,IL-1β,and TNF-α) and hippocampal glutamate (Glu) levels. Furthermore,GLMD restored the positive expression of 5-hydroxytryptamine (5-HT),inhibited the positive expression of ionized calcium-binding adapter molecule 1 (IBA-1) and glial fibrillary acidic protein (GFAP) in the hippocampus,and upregulated the protein expression of phosphorylated PI3K and AKT. In conclusion,GLMD likely exerts sedative and neuroprotective effects by modulating the PI3K/ AKT signaling pathway and inhibiting neuroinflammation.

Key words:

"> Guizhi Jia Longgu Muli Decoction, insomnia, neuroinflammation, PI3K/ AKT signaling pathway, network pharmacology