Network Pharmacology-Based Elucidation of Gu’s Anshen Decoction in Modulating TNF Signaling to Ameliorate Insomnia

Objective: To investigate the therapeutic mechanisms of Gu’s Anshen decoction (GAD) against insomnia (INS), focusing on the regulation of tumor necrosis factor (TNF)-mediated inflammatory pathways. Methods: Bioactive compounds and potential targets of GAD were identified via network pharmacology and constructed herb–compound–target interaction network. INS-related targets derived from peripheral blood mononuclear cells in the Gene Expression Omnibus (GEO) database were intersected with GAD targets to establish a protein-protein interaction (PPI) network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to identify the key pathways. Experimental validation employed a para-chlorophenylalanine (PCPA)-induced INS rat model, integrating behavioral tests, histopathology, and inflammatory cytokine quantification. Results: Network pharmacology revealed multi-target, multi-pathway actions of GAD against INS, with the TNF signaling pathway identified as a critical axis. Animal experiments demonstrated that GAD significantly improved sleep architecture in INS rats, reduced serum and hippocampal levels of TNF-α and interleukin-1β (IL-1β), and downregulated phosphorylated NF-κB p65 (p-p65) expression, indicating suppression of TNF-driven neuroinflammation. Conclusion: GAD ameliorates INS in a rat model by inhibiting TNF-mediated inflammatory signaling, offering a potential mechanistic basis for its therapeutic use in INS management.