Integrated multi-technology exploration of the mechanism by which Badushengji San regulates core targets in diabetic foot ulcer.

Diabetic Foot Ulcer (DFU), a prevalent and refractory diabetic complication, lacks clear pathological targets and effective therapeutic strategies. This study aimed to identify potential DFU biomarkers and preliminarily explore the mechanism of Badushengji San (BDS) in treating DFU. Transcriptome datasets (GSE199939, GSE134431) and single-cell RNA sequencing (scRNA-seq) data (GSE165816) were retrieved to screen differentially expressed genes (DEGs) and DFU-related cell subsets. Intersection genes among DEGs, drug targets, and high-dimensional Weighted Gene Co-Expression Network Analysis (hdWGCNA) genes were then identified. Core biomarkers were subsequently screened using Least Absolute Shrinkage and Selection Operator (LASSO), Support Vector Machine-Recursive Feature Elimination (SVM-RFE), and Boruta algorithms. Macrophages were defined as DFU-associated core cells. Three biomarkers (CYCS, HMOX1, UPP1) showed diagnostic value (the area under the curve (AUC) of UPP1 = 0.973). BDS (50 ng/mL) reversed 40 mM glucose-induced L929 cell injury by restoring HMOX1 and suppressing UPP1 expression. These biomarkers were enriched in pathways like nuclear factor-kappa B (NF-κB), and their abnormal expression was closely linked to DFU pathological progression. Notably, Benzoyl paeoniflorin had the lowest binding energy (-10.06 kcal/mol) with UPP1. Collectively, this study identified CYCS, HMOX1, and UPP1 as potential biomarkers for DFU and preliminarily revealed the potential regulatory effect and molecular characteristics of BDS in DFU intervention, which lays a bioinformatic foundation and provides preliminary experimental clues for further in vivo mechanism research.