Network Pharmacology and Experimental Validation of a Securidaca inappendiculata Xanthone Fraction Targeting Rheumatoid Arthritis via the PI3K/AKT/mTOR/HIF-1α/GSK-3β Axis

Background: Securidaca inappendiculata is a traditional medicinal plant used for the treatment of Rheumatoid Arthritis (RA) and related inflammatory conditions, and it possesses significant anti-RA activity. However, the precise molecular mechanisms underlying its therapeutic effects remain to be fully elucidated. Aim: This study aimed to elucidate the therapeutic mechanisms of a Xanthone-enriched Fraction (XRF) from S. inappendiculata against RA by integrating network pharmacology with experimental validation. Methods: Using both an Adjuvant-Induced Arthritis (AIA) rat model and an LPS/IFN-γ- stimulated THP-1 macrophage model, an integrated strategy was implemented. Potential targets and pathways for the xanthones were predicted via network pharmacology and molecular docking. Thereafter, the anti-arthritic efficacy of the xanthone fraction was evaluated in vivo, while its pro-apoptotic effect and regulation of key signaling pathways were investigated in vitro. Results: XRF significantly alleviated joint swelling and synovial inflammation in AIA rats. Mechanistically, it inhibited the PI3K/AKT/mTOR/HIF-1α signaling cascade and reduced phosphorylation of GSK-3β at Ser9, thereby promoting the apoptosis of pro-inflammatory M1 macrophages. Discussion: This work elucidated the anti-RA mechanism of S. inappendiculata: eight xanthones acted on the PI3K/AKT/mTOR/HIF-1α/GSK-3β axis to induce M1 macrophage apoptosis, validating the integrated approach and laying groundwork for RA treatment and natural product research. Conclusion: The present study provides a scientific foundation for further elucidating the mechanisms of S. inappendiculata against RA.