Metabolomics and Network Pharmacology Revealed the Mechanism of Feining Mixture Against Respiratory Syncytial Virus Pneumonia

Respiratory syncytial virus (RSV) is a leading etiological agent of pneumonia, particularly affecting infants under 2 years and elderly populations, with characteristic clinical manifestations including fever, cough and wheezing. Although Feining mixture-a traditional Chinese medicine formulation based on the classical Maxing Shigan Decoction from the 'Treatise on Febrile Diseases'-has shown clinical efficacy in ameliorating symptoms of viral pneumonias including RSV infection, its precise pharmacological mechanisms remain undefined. Through serum metabolomics and network pharmacology approaches, we systematically identified RSV-associated metabolic disturbances and characterised Feining mixture's regulatory effects on these pathological alterations. Our investigations revealed that Feining mixture significantly attenuated pulmonary inflammation, as evidenced by reduced pro-inflammatory cytokine levels in bronchoalveolar lavage fluid and improved histopathological features in RSV-infected mice. Metabolomic profiling identified 49 differentially expressed metabolites and 14 perturbed metabolic pathways. Network pharmacology and molecular docking analyses predicted the involvement of TP53, AKT1 and STAT3 as core targets, suggesting modulation of PI3K/AKT and TNF signalling pathways. These predictions were experimentally validated through qPCR and Wb analyses, which confirmed that Feining mixture's therapeutic effects are mediated through selective inhibition of the PI3K/AKT1 signalling cascade. These findings provide novel mechanistic insights into Feining mixture's anti-RSV activity, establishing its therapeutic potential through multi-target modulation of critical inflammatory and metabolic pathways.