Modulation of Inflammatory and Oxidative Stress Pathways in COPD: Unraveling the Mechanisms of Fenugreek Through Network Pharmacology and Single-Cell RNA Sequencing

Purpose: This study aims to investigate the potential targets and signaling pathways of fenugreek, a key component of Hanchuan Zupa Granule, in treating COPD using network pharmacology and single-cell RNA sequencing. Patients and Methods: Utilizing single-cell RNA sequencing data on lung tissues from COPD patients (GSE173896 dataset) and network pharmacology, we analyzed alterations in cell populations, pseudotime trajectory of alveolar macrophages, intercellular communication networks, and performed functional enrichment and molecular docking of active fenugreek compounds with disease-related targets. Key bioinformatics tools and databases included Seurat, Monocle2, CellChat, Cytoscape, AutoDock Vina, PyMOL, TCMSP, SwissTargetPrediction, and Comparative Toxicogenomics Database. Results: A total of 39,240 high-quality single cells were obtained from COPD patients, leading to the identification of 23 distinct cell clusters, including 13 major immune and epithelial cell types. Notably, the proportion of senescent alveolar macrophages was significantly higher in COPD samples compared to controls, indicating enhanced involvement in inflammation and oxidative stress (P < 0.05). Pseudotime trajectory analysis categorized nine alveolar macrophage subtypes, demonstrating distinct differentiation pathways concerning inflammation, tissue repair, and cellular senescence. Network pharmacology identified 148 overlapping genes between the targets of fenugreek and COPD, with functional enrichment analyses revealing significant associations with "cytokine-mediated signaling" and "oxidative stress response." Molecular docking indicated that bioactive compounds of fenugreek exhibited strong binding affinities to CCL2 and IL1R1, confirming their roles in disrupting inflammatory signalling and limiting oxidative damage. Conclusion: Our integrated analysis suggests that fenugreek in Hanchuan Zupa Granule holds promise as a therapeutic strategy for modulating inflammatory and oxidative pathways in COPD and provides a strong mechanistic hypothesis centered on the IL1R1/CCL2 axis for experimental validation.