Multi-omics analysis reveals mechanisms of Qingying granules in treating porcine warm disease: dosage optimization and systems biology insights

BACKGROUND: Warm diseases in swine, particularly those associated with highly pathogenic viruses such as African swine fever virus (ASFV) and porcine reproductive and respiratory syndrome virus (PRRSV), inflict devastating economic losses on global pork production. Current control strategies rely heavily on antibiotics and vaccines; however, emerging antimicrobial resistance and vaccine efficacy variability necessitate alternative therapeutic approaches. Traditional Chinese Veterinary Medicine offers potential solutions, yet rigorous evidence-based studies are required to meet modern regulatory standards. This study aimed to investigate Qingying granules, a novel veterinary formulation derived from the classic Qingying decoction, for the treatment of warm disease in pigs and to elucidate its molecular mechanisms through multi-omics analysis. METHODS: A natural disease model was established using pigs presenting with warm disease. A single production batch of Qingying granules was chemically characterized by UPLC-QE-MS and quantified by HPLC for marker compounds (chlorogenic acid and berberine hydrochloride). Animals were randomly allocated to six dosage groups (0.25-2.00 g/kg/day), a positive control (Qingwen Baidu Powder), and a healthy control group (n = 8). Clinical efficacy was assessed by quantitative syndrome scores, body temperature normalization, weight gain, and lung histopathology. Lung tissues from the optimal-dose group, disease control, and healthy control were subjected to transcriptomic sequencing, untargeted metabolomics, and RT-qPCR validation. RESULTS: The optimal therapeutic dose of the Qingying granules was 0.75 g/kg/day, which significantly alleviated fever, reduced clinical signs, and inhibited morphological changes of lung tissue. HPLC analysis of the Qingying granules determined the concentrations of chlorogenic acid and berberine hydrochloride as 0.70 mg/g and 0.77 mg/g, respectively. Multi-omics analysis identified 1,955 differentially expressed genes (DEGs) and 23 altered metabolites between the untreated Disease Model and Healthy Control groups. Furthermore, Qingying granules treatment (0.75 g/kg/day) reversed the expression of key genes (LCN2, LRRC18, PDK4, IL1RL1, S100A4) and normalized critical metabolites (7-hydroxyetodolac). Integrated pathway analysis revealed modulation of the PI3K-Akt signaling pathway, cytokine-cytokine receptor interactions, and glycolysis/gluconeogenesis. CONCLUSION: Qingying granules at 0.75 g/kg/day exerts potent therapeutic effects on porcine warm disease through multi-targeted regulation of inflammation, immune homeostasis, and metabolic balance. These findings establish a scientific foundation for evidence-based clinical application of traditional herbal formulations in antibiotic-free swine production and demonstrate the utility of multi-omics approaches in veterinary pharmacology research.