Spectrum-effect relationship and network pharmacology reveal key anti-rheumatoid arthritis and attenuated toxicity components of processed curculigo rhizome.

ETHNOPHARMACOLOGICAL RELEVANCE: Curculigo rhizome (CR), a traditional Chinese medicine (TCM), has been used for centuries to treat rheumatic conditions and tonify kidney yang. Yellow rice wine processing is a classical preparation method in TCM practice believed to enhance therapeutic efficacy and reduce toxicity, yet its scientific basis remains insufficiently validated.

AIM OF THE STUDY: This study aimed to systematically compare the chemical profiles and anti-rheumatoid arthritis (RA) effects of raw CR (RCR) and wine-processed CR (WCR), establish spectrum-effect relationships to identify key bioactive components, and explore the potential molecular mechanisms through an integrated approach combining chromatographic fingerprinting, chemometric analysis, network pharmacology, and experimental validation.

MATERIALS AND METHODS: HPLC fingerprinting was employed to characterize the chemical constituents of RCR and WCR extracts. The anti-RA effects were evaluated in collagen-induced arthritis (CIA) mice and inflammatory cell models (TNF-α-stimulated MH7A synoviocytes and LPS-induced RAW264.7 macrophages). Orthogonal partial least squares (OPLS) analysis was applied to establish quantitative correlations between chromatographic peaks and pharmacological outcomes. Network pharmacology and molecular docking were conducted to predict potential targets and pathways, followed by experimental verification of glycolysis-related mRNA expression.

RESULTS: HPLC fingerprinting identified 13 common peaks in RCR and WCR, with significant quantitative differences between raw and processed forms. Both RCR and WCR alleviated arthritis severity in CIA mice, as evidenced by reduced paw swelling, arthritis scores, and synovial inflammation, along with decreased serum pro-inflammatory cytokines (TNF-α, IL-1β). Notably, WCR exhibited comparable or potentially improved efficacy to RCR while demonstrating a lower potential for nephrotoxicity. In vitro studies confirmed that both extracts inhibited pro-inflammatory cytokine release, nitric oxide production, and cell migration in activated synoviocytes and macrophages. Spectrum-effect relationship analysis identified peaks 3, 6, 7, 9, and 10 as key candidate markers associated with anti-RA activity. Network pharmacology revealed 157 potential therapeutic targets, with GAPDH, EGFR, CASP3, and SRC identified as hub nodes. Molecular docking predicted favorable binding affinities between active components (orcinol gentiobioside, chlorogenic acid) and core targets. Mechanistically, preliminary evidence indicated that both RCR and WCR suppressed the mRNA expression of glycolytic enzymes (HK2, PKM2, PFKFB3, GLUT-1), suggesting that inhibition of glycolytic reprogramming in inflammatory cells may contribute to their anti-RA effects.

CONCLUSIONS: This study provides scientific evidence supporting the traditional wine-processing method for CR and demonstrates that WCR maintains comparable anti-RA efficacy with potentially improved safety compared to RCR. The identified candidate bioactive components and the potential role of glycolysis inhibitory mechanism offer novel insights for quality control standardization and further investigation of CR-based interventions for rheumatoid arthritis.