Mechanism of Action of Astragalus membranaceus for Treating Diabetic Foot Ulcers Based on Single-Cell RNA Sequencing Data and Network Pharmacology
Summary: Published March 10, 2026 in Scientific Reports (Nature), this open-access study from Kunming University of Science and Technology and affiliated hospitals in Yunnan, China integrates single-cell RNA sequencing (scRNA-seq) and network pharmacology to elucidate the molecular mechanisms by which Astragalus membranaceus (AM, Huangqi) — a widely used traditional Chinese medicinal herb — may modulate the pathological wound microenvironment in diabetic foot ulcers (DFUs). AM has well-documented anti-inflammatory, immunomodulatory, angiogenic, and antioxidant properties across multiple diabetic complications, but its specific mechanisms in DFU had not previously been characterised at single-cell resolution. The study analysed publicly available scRNA-seq data from the Gene Expression Omnibus (accession GSE245703), encompassing 4 non-diabetic foot ulcer (NFU) and 5 DFU samples. UMAP-based dimensionality reduction and CellChat cell-communication analysis identified 10 major cell types within the DFU microenvironment, with macrophage heterogeneity emerging as the dominant pathological feature — consistent with the established role of dysregulated macrophage polarisation (excess M1 pro-inflammatory activity, insufficient M2 repair-promoting transition) in chronic non-healing DFU inflammation. Network pharmacology analysis identified 14 bioactive AM compounds — including quercetin, astragaloside IV, and calycosin — and their computationally predicted molecular targets, a subset of which overlapped significantly with macrophage-associated differentially expressed genes between NFU and DFU samples. Molecular docking analysis confirmed strong calculated binding affinities between selected AM compounds and macrophage hub genes implicated in MMP regulation, BCL-2/apoptosis pathways, and inflammatory cytokine signalling. Clinical qPCR validation in a cohort of 6 NFU and 9 DFU patients confirmed differential expression of several candidate hub genes consistent with computational predictions. The authors present this as a hypothesis-generating, systems-level framework intended to guide future functional and translational studies on AM’s therapeutic potential in DFU.
Key Highlights:
- Integrative design: scRNA-seq (GSE245703; 4 NFU + 5 DFU) + network pharmacology + molecular docking + clinical qPCR validation (n=15); Yunnan University of Science and Technology, China
- scRNA-seq: 10 cell types identified in DFU microenvironment; macrophage heterogeneity is dominant — excess M1 polarisation and insufficient M2 transition characterises chronic DFU inflammation
- 14 bioactive AM compounds identified via SwissADME pharmacokinetic screening, including quercetin, astragaloside IV, and calycosin — each with established anti-inflammatory and immunomodulatory activity
- Network pharmacology: predicted AM compound targets overlap with macrophage-associated DEGs in DFU vs. NFU — particularly genes regulating MMP activity, apoptosis pathways, and pro-inflammatory cytokine signalling
- Molecular docking: strong calculated binding affinities between AM bioactive compounds and macrophage hub genes — supports plausibility of the predicted therapeutic interaction
- qPCR validation (6 NFU, 9 DFU): differential expression of candidate hub genes confirmed in clinical samples; study is explicitly hypothesis-generating — functional in vitro/in vivo validation studies are required before clinical translation
Keywords: Astragalus membranaceus wound healing, diabetic foot ulcer macrophage, single cell RNA sequencing wound, network pharmacology wound care, traditional Chinese medicine DFU, macrophage polarization diabetic wound
Xia Li Yan Dong Chong Huang Guozhong Zhou Yanjie Ning Yuru Liu Ruqin Zhang Ying Yang Nan Chen