Deciphering the therapeutic mechanisms of Qingfei Litan prescription in acute lung injury: an integrated framework of UHPLC–MS/MS, systems pharmacology, machine learning, and in vivo validation

Background Acute lung injury (ALI) is primarily defined by hyper-inflammation and the breakdown of the alveolar-capillary barrier, frequently culminating in refractory respiratory failure. Current clinical interventions—predominantly glucocorticoids and supportive therapies—often yield suboptimal outcomes and are associated with significant side effects. Qingfei Litan Prescription (QFLTP), a traditional Chinese medical prescription indicated for inflammatory respiratory disorders, exhibits significant therapeutic potential; however, the precise molecular mechanisms driving its protective effects are currently under-investigated. Methods Qualitative detection of the QFLTP ingredients was accomplished via UHPLC-MS/MS analysis. Active ingredients and their potential targets were screened using online prediction tools and traditional Chinese medicine databases, Concurrently, genes associated with ALI were retrieved via GeneCards, DisGeNET, and OMIM repositories. The intersection of these target sets underwent rigorous investigation, encompassing protein-protein interaction (PPI) mapping, alongside Gene Ontology (GO) and KEGG pathway enrichment evaluations, and multiple data-driven algorithms to identify hub genes. Computational modeling and simulation methods were performed to evaluate ligand-target binding. An LPS-induced ALI mouse model was established for in vivo validation. Histopathological examination, bronchoalveolar lavage fluid (BALF) protein quantification, inflammatory cytokine assays, arterial blood gas analysis, immunohistochemistry, immunofluorescence, Western blot, and qRT-PCR served to evaluate the therapeutic potential of QFLTP. Results A total of 47 chemical constituents and 298 overlapping therapeutic targets were identified. Integrated computational and data-driven analyses highlighted POLB, FGF1, and ARG1 as potential hub genes involved in QFLTP-mediated protection against ALI. GO and KEGG enrichment analyses revealed that pharmacological restorative capacity of QFLTP activity demonstrated a robust functional linkage to inflammatory responses, PI3K-Akt signaling, VEGF signaling, and immune-related pathways. In vivo trials revealed that QFLTP administration markedly mitigated pulmonary histopathological impairment while simultaneously suppressing the secretion of pro-inflammatory mediators and pulmonary edema, improving lung function and arterial blood gas parameters, and promoted the recovery of junctional protein expression of ZO-1, ZO-2, and VE-cadherin in lung tissues. In addition, qRT-PCR profiling revealed that QFLTP treatment effectively normalized the aberrant mRNA levels of POLB, FGF1, and ARG1 in the lung tissues of the ALI cohort. Conclusion QFLTP exerts protective effects against ALI via its multi-component, multi-target, and multi-pathway mechanisms, potentially by suppressing inflammatory responses, restoring pulmonary barrier integrity, and regulating the expression of hub genes including POLB, FGF1, and ARG1.