[A network pharmacology-based study to explore the mechanism of a novel sleep-aiding soup for the treatment of insomnia by modulating microglia activation].

Objective Building on prior clinical evidence demonstrating the efficacy of a novel sleep-aid decoction in treating insomnia among elderly patients, this study aimed to explore its potential mechanism of action by integrating network pharmacology with in vitro experiments, with a focus on microglial activation regulation. Methods Active compounds and potential targets of the decoction were identified using the TCMSP and HERB databases. These targets were then intersected with insomnia-and microglial activation-related targets to construct a protein-protein interaction (PPI) network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted via the DAVID platform to identify key signaling pathways. An in vitro model of microglial activation was established by stimulating BV2 cells with lipopolysaccharide (LPS). The effects of the decoction-containing serum on inflammatory cytokine levels and microglial activation markers were assessed using ELISA, quantitative real-time PCR (qRT-PCR), and nitric oxide (NO) assays. Results A total of 87 active compounds and 831 predicted targets were identified, including 62 overlapping targets associated with insomnia and microglial activation. PPI and pathway analyses indicated significant enrichment in classical neuroinflammation-related pathways, including the advanced glycation end product-receptor for advanced glycation end product (AGE-RAGE), Toll-like receptor (TLR), Janus kinase/signal transducer and activator of transcription (JAK/STAT), and phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathways. In vitro experiments showed that the decoction-containing serum significantly suppressed LPS-induced expression of interleukin 1β (IL-1β), IL-6, tumor necrosis factor α (TNF-α), and NO, while downregulating both mRNA and protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2). These results indicated effective inhibition of microglial activation and inflammatory responses. Conclusion The novel sleep-aid decoction may alleviate insomnia by modulating the AKT signaling pathway, thereby suppressing proinflammatory microglial polarization and reducing neuroinflammation. This study reveals a potential immunomodulatory mechanism of this traditional Chinese medicine formula and highlights the value of its multi-component, multi-target approach for sleep disorder management. Further studies integrating multi-omics and single-cell technologies are warranted to elucidate the specific roles and targets of its key active components.