Integrated network pharmacology, metabolomics, and proteomics analysis of Shiqi Waigan Granules for treating exogenous febrile disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Fever, which is closely associated with inflammation, is classified in Traditional Chinese Medicine as either exogenous or endogenous. Shiqi Waigan Granules (SWG) are indicated for treating fever and headache caused by exogenous factors, as well as for preventing influenza.

AIM OF THE STUDY: To investigate the main therapeutic compounds and potential mechanisms of SWG in treating exogenous febrile disease.

MATERIALS AND

METHODS

UPLC-QTOF/MS profiling identified the absorbed compounds of SWG in rat plasma. Network pharmacology, metabolomics, and proteomics were employed to pinpoint the targets and pathways, while molecular docking, immunofluorescence, and Western blot validated these findings.

RESULTS

A total of 47 prototype compounds and 13 metabolites were detected in rat plasma. Network analysis identified 145 overlapping targets associated with antipyretic and anti-inflammatory GO and KEGG pathways. Molecular docking revealed that key bioactive compounds-Asprelloside B, Verbascoside, Isoverbascoside, Chlorogenic acid, and Ellagic acid 4-O-xylopyranoside-exhibited binding energies lower than -7.0 kcal mol-1 with core proteins PIK3CA, AKT1, NF-κB, and IκB-α. SWG significantly reduced LPS-induced fever in rats, an effect likely mediated by suppression of pro-inflammatory cytokine production. Furthermore, SWG modulated pivotal metabolites identified through metabolomics, influencing the thermoregulatory center and exerting antipyretic effects. Key metabolic pathways involved included unsaturated fatty acid metabolism, lipid metabolism, and arachidonic acid metabolism. Proteomic analysis revealed differentially expressed proteins enriched in the NF-κB and PI3K/Akt signaling pathways. Western blot and immunofluorescence assays confirmed that SWG inhibited inflammatory activation by regulating the PI3K/Akt and NF-κB pathways.

CONCLUSION

SWG exerts multi-compound, multi-target, multi-pathway control of exogenous febrile disease, offering a rational basis for its clinical application and novel strategies for fever management.