Mechanisms of asiaticoside in keloid treatment: Insights from a single-cell transcriptomic approach combined with network pharmacological analysis

Abstract

This work sought to clarify the molecular processes through which asiaticoside operates in treating keloid by utilizing an integrated approach employing scRNA-seq and systems pharmacological methods. We performed scRNA-seq on three patients' keloid and adjacent normal skin samples. We detected 1500 highly variable genes and further examined them using PCA and tSNE techniques. Drug targets of asiaticoside were predicted through multiple databases, and overlapping genes with keloid-related cell types (macrophages and keloid fibroblasts (KFs)) were analyzed. GO and KEGG enrichment evaluations were performed, followed by in vitro assays to confirm the influence of asiaticoside on macrophage and keloid fibroblast. scRNA-seq revealed significant cellular heterogeneity between keloid and adjacent tissue, with macrophages and KFs identified as major contributors to its pathogenesis. Network pharmacology identified NF-κB, PI3K-Akt signaling, etc., through which asiaticoside exerts its anti-inflammatory and anti-fibrotic effects. In vitro studies confirmed that asiaticoside inhibited macrophage activation and reduced the pro-fibrotic effects of macrophage on KF. Our study demonstrates asiaticoside alleviates keloid by modulating macrophage function and KF activity through NF-κB and PI3K-Akt signaling. These findings provide a novel mechanistic understanding of asiaticoside’s therapeutic effects and offer insights into its clinical application.