Ze-qi decoction inhibits neutrophil extracellular trap formation to suppress the growthand metastasis of NSCLC

This study aimed to explore the material basis for anit-NSCLC efficacy of Ze-qi decoction (ZQD) and elucidate the molecular mechanisms. UPLC-HRMS was used to identify the primary chemical components. The potent molecular mechanisms were analyzed using network pharmacology. A subcutaneous tumor mouse model was established. The subcutaneous tumor was stripped for transcriptomic analysis. 253 DEGs were identified in the ZQD-H versus Model, including 167 upregulated and 86 downregulated genes. The heatmap demonstrated that the gene expression patterns differed significantly between the two groups. Functional exploration was conducted using GO and KEGG analyses. Most entries were immune-associated, such as the regulation of tumor necrosis factor and cytokine production, as well as immune receptor activity. From the KEGG enrichment analysis, we identified 29 significant KEGG pathways mostly associated with cancer, immunology, and metabolism, which corresponded to the results of the GO analysis. The neutrophil extracellular traps (NETs) formation was uncovered as the anti-NSCLC target of ZQD by pharmacological research and transcriptomic profile. The molecular docking analysis was conducted to predict the interaction of 16 core components with proteins HIF1A, CD18, and MPO that were jointed to neutrophil recruitment and NETs formation.The underlying molecular mechanisms of ZQD against NSCLC were investigated by western blot, immunofluorescence, immunohistochemistry, and Elisa assays. ZQD hindered the neutrophils recruitment and activation by decreasing HIF-1α, CD18, and ICAM-1 levels and suppressed NETs formation shown as the downregulated biomarkers (CitH3, MPO, and NE). Finally, the lung metastasis mouse model was constructed dor further validation. Fewer nodules were observed on the surface of the lungs in mice treated with ZQD. NETs-related proteins CitH3 and Ly6G, as well as MPO were less expressed in the lungs after ZQD intervened in comparison of model group.