Investigation of the mechanism of Xiaoxianxiong Tang in ameliorating pneumonia induced by graphene oxide through network pharmacology, molecular docking, molecular dynamics simulation, and experimental validation

• CASP3, SRC, STAT3, EGFR, and AKT1 can bind to vitamin-e spontaneously. • Xiaoxianxiong Tang can inhibit GO-induced inflammation by reducing the levels of IL-6, IL-1β, and TNF-α. • Xiaoxianxiong Tang can inhibit GO-induced inflammation by reducing the mRNA levels of CASP3, SRC, STAT3, EGFR, and AKT1. Graphene oxide (GO) can lead to inflammation of the lung. Xiaoxianxiong Tang (XXXT) has been used to treat pneumonia. However, its mechanisms of action are unclear. In this study, we explored the molecular mechanism of XXXT in pneumonia using network pharmacology, molecular docking, molecular dynamics (MD) simulation, and experimental validation. Network pharmacology approaches were employed to screen the active compounds and targets of XXXT, and the targets of pneumonia. Molecular docking combined with MD simulations was used to predict the modes, affinities, and long-term stability of binding between compounds and targets. Subsequently, the expression levels of interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α) and mRNA expression of caspase-3 (CASP3), steroid receptor coactivator (SRC), signal transducer and activator of transcription 3 (STAT3), epidermal growth factor receptor (EGFR), and serine/threonine kinase 1 (AKT1) were measured. Molecular docking and MD simulations showed that the main active compounds possessed high binding affinity to the core targets. Experiments showed that XXXT alleviated GO-induced inflammation through decreasing inflammatory cytokine levels as well as downregulating mRNA expression of CASP3, SRC, STAT3, EGFR, and AKT1. Our results elucidated the active compounds, potential therapeutic targets, and molecular mechanisms of XXXT in alleviating pneumonia, which present a new strategy for the future alleviation of pneumonia by TCM.