Vestitol Ameliorates Ferroptosis-Associated Features and Osteoarthritis Progression Through Modulation of the GSK3B/Nrf2/GPX4 Pathway.

This study aimed to investigate whether vestitol ameliorates osteoarthritis (OA) and the mechanisms involved. We employed destabilization of the medial meniscus (DMM)-constructed OA mouse model and IL-1β-induced chondrocyte model to evaluate vestitol's effects on OA. In vivo, pathological alterations in cartilage tissues were assessed via safranin O and H&E staining. In vitro, chondrocyte viability and apoptosis were detected utilizing CCK-8 and flow cytometry. Vestitol's mechanisms in OA were explored using network pharmacology analysis. Factors related to extracellular matrix (ECM) degeneration, inflammation, oxidative stress, ferroptosis, and pathway were checked using western blot, RT-qPCR, immunohistochemistry, and corresponding commercial reagent kits. In OA mice model, vestitol decreased cartilage destruction and OARSI score. Vestitol enhanced the levels of COL2A1, Aggrecan, SOD, FTH1, but reduced the levels of MMP-13, ADAMTS5, IL-6, IL-1β, COX-2, TNF-α, MDA, Fe2+, ROS, ACSL4 and 4-HNE. In IL-1β-induced chondrocyte model, vestitol enhanced cell viability and inhibited cell apoptosis. The changes in indicators related to ECM degradation, inflammation, oxidative stress, and ferroptosis were consistent with in vivo experiments. Moreover, vestitol increased p-GSK3B (Ser9), Nrf2, and GPX4 levels both in vivo and in vitro. The inhibition of vestitol on chondrocyte apoptosis, ECM degradation, inflammation, and ferroptosis-associated features was reversed by GSK3B activation. Vestitol mitigates OA by suppressing chondrocyte apoptosis, ECM degradation, inflammation, and ferroptosis-associated features, with pharmacological evidence implicating the PI3K/AKT/GSK3B axis and downstream modulation of the Nrf2/GPX4 pathway.