Integrating Metabolomics and Network Pharmacology: Investigating the Therapeutic Mechanism of Atractylodes Rhizome Against Rheumatoid Arthritis

<b>Purpose:</b> The purpose of this study is to investigate the bioactive constituents of Atractylodes Rhizome (AR) and to explore its mechanism of action in the treatment of rheumatoid arthritis (RA). <b>Methods:</b> The research mainly adopts the methods of tissue metabolomics and network pharmacology. Firstly, we employed a metabolomics strategy to obtain the metabolite profile and utilized PCA/OPLS-DA analyses to identify the differential metabolites involved in the treatment of RA by AR. Subsequently, we determined the key target metabolic pathways of AR in RA treatment. Next, a network pharmacology approach was employed to identify active compounds, potential targets, and signaling pathways for AR in RA treatment, with a PPI network constructed. These predictions were then validated through molecular docking simulations, followed by in vivo verification using a CFA-induced RA rat model. The anti-RA efficacy was evaluated through synovial histopathology and cytokine assays, with the key mechanistic insights being confirmed at the molecular level by RT-qPCR and WB. <b>Results:</b> The results of the metabolomics study showed that AR regulated 28 differential metabolites linked to glycerophospholipid, linoleic acid, and alpha-linolenic acid metabolism. Network pharmacology identified Wogonin, Atractyloyne, and Atractylenolide II as key active compounds, acting through pathways such as Pathways in cancer and PI3K-Akt signaling, combined with the metabolites to jointly analyze the metabolic pathways, and were verified by correlation analysis. Molecular docking confirmed the main active ingredients' strong binding to core targets. In AIA rats, AR treatment reduced synovial inflammation and lowered serum levels of IL-6 and MMP-9. At the molecular level, AR up-regulated Bcl-2, down-regulated Bax, and inhibited the SRC/JAK2-STAT3 pathway by decreasing EGFR, SRC, JAK2, and p-STAT3 expression. <b>Conclusion:</b> These findings may illuminate the mechanism by which Atractylodes Rhizome exerts its effects via the JAK2/SRC-STAT3 axis, thereby revealing its potential mechanism of action against rheumatoid arthritis.