An integrated approach combining transcriptomics, network pharmacology, and experimental validation are used to investigate the mechanism of Lonicerae Japonicae Flos and Forsythiae Fructus extracts against acute lung injury in chickens.

ETHNOPHARMACOLOGICAL RELEVANCE: Acute lung injury (ALI) is a severe respiratory disease characterized by immune dysregulation and inflammation. In the poultry farming industry, bacterial pathogens are the primary cause of respiratory health issues in poultry. Severe Gram-negative bacterial infections leading to endotoxemia can easily result in ALI. Currently, there are limited effective treatment options for ALI. Lonicerae Japonicae Flos and Forsythiae Fructus are a classic and widely used herbal pair in traditional Chinese medicine (TCM), with a long-standing clinical application for treating heat-toxin-related respiratory diseases, fever, cough, and inflammatory syndromes. This classic combination is documented to clear heat, detoxify, and relieve lung inflammation, matching the pathological features of bacteria-induced ALI in chickens. Despite its long-term ethnopharmacological use for respiratory disorders, the specific mechanism underlying its protective effect against LPS-induced ALI in poultry remains largely unclear. This study is the first to integrate UPLC-MS/MS, transcriptomics, network pharmacology, molecular docking, and in vivo experiments to reveal that LF alleviates LPS-induced ALI in broilers by suppressing MAPK signaling and regulating apoptosis. Elucidate the pharmacodynamic material basis and mechanisms of action of the Lonicerae Japonicae Flos and Forsythiae Fructus extracts (LF) in the context of lipopolysaccharide (LPS)-induced ALI, providing a theoretical foundation for the use of this herbal pair in improving ALI.

MATERIALS AND METHODS: Broilers were used to investigate the protective effects and potential mechanisms of the LF against LPS-induced ALI. The bioactive compounds in the LF were analyzed using UPLC-MS/MS. Histopathological evaluation of lung tissues was conducted using hematoxylin and eosin (H&E) staining. ELISA was conducted to measure the levels of inflammatory factors in the serum and inflammatory and oxidative stress markers in the lung tissue. Network pharmacology, transcriptomics, and molecular docking techniques were employed to investigate the therapeutic mechanisms of the LF on ALI.

RESULTS: LF significantly improved pathological damage in the lungs of chickens, reduced the degree of pulmonary edema, and modulated the levels of inflammatory factors and antioxidant enzyme activity. This combination reduced inflammatory responses and enhanced antioxidative capacity in the chickens. Bioinformatics analysis of transcriptomic data highlighted MAP kinase activity, antioxidant activity, and inflammation control as key roles of LF in treating ALI. Combined UPLC-MS/MS, network pharmacology, and molecular docking analyses revealed that luteolin, β-sitosterol, and phillyrin may treat ALI by regulating the MAPK signaling pathway and apoptosis, and these were identified as potential active compounds.

CONCLUSIONS: This study confirmed that the LF improved LPS-induced ALI in chickens by inhibiting the activation of the MAPK signaling pathway and regulating apoptosis. Luteolin, β-sitosterol, and phillyrin were identified as key active compounds, providing a theoretical basis for further research.