Pharmacological activation of SIRT1-AMPK by ginsenoside Rb1: a novel therapeutic strategy for pressure injury via dual suppression of ferroptosis and inflammation.

BACKGROUND: Pressure injuries (PIs) are a major clinical problem, and current treatments offer limited efficacy. Ferroptosis-driven oxidative damage and chronic inflammation severely impair wound healing. Ginsenoside Rb1 (Rb1), a bioactive component of Panax ginseng, possesses antioxidant and anti-inflammatory activities, yet its therapeutic potential in PI via ferroptosis regulation has not been investigated. This study aims to determine whether Rb1 promotes PI wound repair by activating the Sirtuin 1-AMP-activated protein kinase (SIRT1-AMPK) pathway to inhibit ferroptosis and inflammation, thereby providing a new pharmacological strategy for PI management. METHOD: Transcriptomic profiling was performed on dorsal skin tissues from normal rats, pressure injury rats, and Rb1-treated rats using RNA sequencing to identify differentially expressed genes (DEGs), followed by Gene Ontology (GO)/Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, network pharmacology analysis, and protein-protein interaction (PPI) network construction to screen potential regulatory pathways. In vitro, ferroptosis was induced in an L929-HaCaT co-culture system using erastin/RSL3, and cells were treated with various concentrations of Rb1. Cell viability, reactive oxygen species (ROS) levels, ferroptosis-related markers (GPX4, SLC7A11, and ACSL4), and SIRT1-AMPK pathway proteins were evaluated by using the CCK-8, assay, fluorescence assays, Western blotting, and RT-qPCR. In vivo, a PI model was created in Sprague-Dawley (SD) rats, followed by administration of Rb1. Wound healing, histopathology, oxidative stress indices, inflammatory cytokines, and SIRT1-AMPK activation were assessed. RESULTS: Integrated transcriptomic and network pharmacology analyses identified the SIRT1-AMPK axis as a key mediator of Rb1-induced wound repair. In vitro, Rb1 dose-dependently attenuated erastin/RSL3-induced ferroptosis, decreased ROS levels, and increased the expressions of GPX4, SLC7A11, and ACSL4, while simultaneously activating SIRT1 and downstream p-AMPK. Rescue experiments showed that blocking the activities of SIRT1 or AMPK diminished the protective effects of Rb1 against ferroptosis. In vivo, high-dose Rb1 accelerated wound closure, activated SIRT1-AMPK signaling, enhanced ferroptosis-related protein expression, and reduced tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels. CONCLUSION: Rb1 functions as an SIRT1-AMPK activator that inhibits ferroptosis and inflammation to promote PI wound healing. These findings underpin the efficacy of Rb1 as a promising multi-target therapeutic candidate for future clinical development.