Integrating Seropharmacology, Network Pharmacology, and Single‐Cell Transcriptomics to Decipher the Dual Anti‐Tumor Mechanisms of Astragalus– Curcuma zedoaria in Lung Squamous Cell Carcinoma

This study sought to elucidate the antitumor mechanisms of the traditional herbal combination Astragalus-Curcuma zedoaria (A-C) in lung squamous cell carcinoma (LUSC). Drug-containing serum was prepared from mice administered an A-C decoction, and serum-absorbed bioactive components were identified using high-resolution mass spectrometry. To investigate potential targets and pathways within the tumor immune microenvironment, network pharmacology analysis was integrated with bulk transcriptomic data from TCGA-LUSC and single-cell RNA sequencing (scRNA-seq). The analytical methodologies employed included differential gene expression analysis, weighted gene co-expression network analysis (WGCNA), functional enrichment analysis, and molecular docking. The scRNA-seq data identified 22 distinct cell clusters, with correlation analysis suggesting that A-C primarily exerts its effects through vascular endothelial cells. Among the serum-derived components, demethoxycurcumin-a principal curcuminoid from C. zedoaria-demonstrated the highest binding affinity with the TEK receptor tyrosine kinase (-7.5 kcal/mol), indicating its potential role in modulating endothelial function and vascular-immune interactions. In conclusion, the A-C combination exhibits a synergistic effect in lung squamous cell carcinoma (LUSC) by modulating differentially expressed genes, with demethoxycurcumin identified as a pivotal bioactive compound targeting TEK. These results elucidate novel dual antitumor mechanisms of the A-C herbal pair, achieved through the regulation of the vascular-immune microenvironment.