Ginsenoside Rg1 delays the senescence of adipose-derived stem cells: network pharmacology and experimental validation

BACKGROUND: Mesenchymal stem cell (MSC) senescence limits their therapeutic potential. Adipose-derived stem cells (ADSCs), though easily accessible, are prone to senescence under oxidative or inflammatory stress. Ginsenoside Rg1, with antioxidant and anti-inflammatory properties, may counteract this process. This study investigates whether Rg1 can alleviate ADSC senescence and its underlying mechanisms. METHODS: To explore the molecular mechanisms by which Ginsenoside Rg1 mitigates ADSC senescence, network pharmacology and molecular docking were applied to identify potential signaling pathways and targets. Cell viability was measured using the Cell Counting Kit-8 (CCK-8) assay to select the optimal modeling concentration. Flow cytometry was used to analyze cell cycle distribution and immune-related marker expression in ADSCs. Reverse transcription quantitative PCR (RT-qPCR) quantified stemness- and senescence-related gene expression. Immunofluorescence (IF) staining assessed senescence-associated protein levels. Western blotting (WB) was performed to examine key proteins in the PI3K/AKT signaling pathway. RESULTS: Network pharmacology identified 22 aging-associated genes targeted by Rg1, with AKT1 emerging as a central hub protein exhibiting high binding affinity. In hydrogen peroxide (H₂O₂)-induced senescence model, Rg1 treatment significantly enhanced cell viability and stemness, while reducing senescence markers p16 and p21. Furthermore, Rg1 attenuated DNA damage, decreased pro-inflammatory cytokine expression, and restored telomere length. It also promoted cell cycle progression and upregulated immune checkpoint proteins CD59 and CD73. Mechanistic analysis revealed that these anti-senescent effects are mediated via activation of the PI3K/AKT signaling pathway. CONCLUSION: Rg1 effectively attenuates ADSC senescence by activating the PI3K/AKT signaling pathway, promoting cell cycle progression and enhancing immune evasion capacity. This reveals a novel anti-aging mechanism and offers new insights for MSC-based regenerative therapies, while also providing modern evidence supporting the traditional Jing-Qi theory.