Integrative network pharmacology and in vitro/in vivo validation reveal the protective effects of sotetsuflavone against osteoarthritis associated with PI3K/Akt/NF-κB signaling.

Authors

Liang Chen, Yi'nan Pan, Xia Cao, Wenfeng Peng, Chang Zhu, Jing Ye, Zengru Xie

Published in

Scientific reports. Jul 03, 2026. Epub Jul 03, 2026.

Abstract

Osteoarthritis (OA) is a prevalent whole-joint disease characterized by progressive cartilage degeneration, extracellular matrix (ECM) breakdown, synovial inflammation, and pathological changes in multiple joint tissues. Inflammatory signaling plays a central role in chondrocyte catabolic activation and cartilage matrix degradation. Sotetsuflavone (SF), a naturally occurring biflavonoid isolated from Cycas species and other medicinal plants, has been reported to possess anti-inflammatory and antioxidant properties; however, its protective effects and related molecular mechanisms in OA remain unclear. This study aimed to investigate whether SF attenuates OA-associated inflammatory and catabolic responses and to explore the functional involvement of phosphoinositide 3-kinase/protein kinase B/nuclear factor-κB (PI3K/Akt/NF-κB) signaling in its protective effects. An integrated strategy combining network pharmacology, molecular docking, and experimental validation was used. Potential SF-related targets and OA-associated genes were collected from public databases, and overlapping targets were analyzed by protein-protein interaction (PPI) network construction and functional enrichment analyses. Molecular docking was performed to evaluate the predicted binding modes between SF and selected pathway-related proteins. Primary mouse chondrocytes stimulated with interleukin-1β (IL-1β) were used to assess the effects of SF on inflammatory mediator production, ECM metabolism, and PI3K/Akt/NF-κB signaling. To further evaluate pathway involvement, a rescue experiment was performed using 740Y-P, a PI3K/Akt pathway activator. A destabilization of the medial meniscus (DMM)-induced mouse model was established to evaluate the protective effects of SF against OA-related structural and inflammatory changes in vivo. Network pharmacology analysis identified 68 overlapping targets between SF and OA-associated genes, with enrichment mainly involving inflammation-related and kinase-dependent pathways, including PI3K/Akt and NF-κB signaling. Molecular docking revealed favorable binding affinities of SF toward key targets. In vitro, SF reduced the expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) and decreased the production of nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). SF also alleviated IL-1β-induced ECM catabolism, as shown by reduced matrix metalloproteinase-13 (MMP-13) and a disintegrin and metalloproteinase with thrombospondin motifs-5 (ADAMTS-5) levels and partial preservation of type II collagen (Col II) and aggrecan. Mechanistically, SF decreased PI3K/Akt phosphorylation, restored Inhibitor of nuclear factor-κB alpha (IκBα) expression, and reduced nuclear accumulation of NF-κB p65. Importantly, activation of PI3K/Akt signaling by 740Y-P partially reversed the inhibitory effects of SF on NF-κB activation and inflammatory mediator production, supporting the functional involvement of the PI3K/Akt/NF-κB axis. In vivo, SF administration alleviated cartilage destruction, reduced Osteoarthritis Research Society International (OARSI) scores, decreased MMP-13 expression, restored Col II expression, and lowered the expression of pro-inflammatory cytokines in joint tissues. These findings suggest that SF attenuates OA-associated inflammatory activation and cartilage matrix degradation in experimental models. The protective effects of SF are at least partly associated with suppression of the PI3K/Akt/NF-κB signaling axis. Further studies are warranted to clarify direct target engagement, pharmacokinetics, long-term safety, optimized delivery strategies, and therapeutic efficacy in more comprehensive OA models.

PMID:
42399461
Bibliographic data and abstract were imported from PubMed on 04 Jul 2026.

Read full publication at:
Please sign in to see all details.

Stats

1-terrible, 9-excellent. How would you rate this publication? Sign in in to submit your rating.

Post a comment

You need to be signed in to post comments. You can sign in here.

Comments

There are no comments yet.