Ganoderic acid L attenuates LPS-induced inflammatory lung injury through integrated network pharmacology, transcriptomics and target-engagement analyses

Severe burns can cause secondary pulmonary inflammation driven by endotoxin-related inflammatory activation and oxidative stress. Here, we integrated untargeted LC-MS, network pharmacology, transcriptomics, molecular docking, molecular dynamics simulation and target-engagement assays to identify and validate a candidate active constituent from Ganoderma lucidum . LC-MS identified 660 compounds, with terpenoids as a predominant class. Network pharmacology prioritized inflammatory signaling pathways related to PI3K-Akt, MAPK and NF-κB, and integration with RNA-seq identified 92 transcriptomics-supported candidate targets. Ganoderic acid L showed favorable predicted binding to PIK3CA and was selected for experimental validation. In an LPS-induced inflammatory lung injury mouse model, ganoderic acid L alleviated histopathological injury, pulmonary edema and bronchoalveolar lavage fluid protein leakage; reduced TNF-α, IL-6, IL-1β, MPO and MDA levels; and restored SOD and GSH-Px activities. RT-qPCR and western blotting indicated suppression of inflammatory gene expression and modulation of Akt/NF-κB/MAPK signaling. Surface plasmon resonance showed micromolar-level interaction with PI3K, and CETSA provided supportive evidence for potential target engagement. These findings identify ganoderic acid L as a candidate anti-inflammatory triterpenoid for LPS-induced lung injury, while causal PI3K engagement requires further validation.