Extraction, Purification of C‑Glycosides from Arisaema decipiens for Studying its Anti-Inflammatory and Bioactivities

Arisaema decipiens-(A. decipiens), a traditional Chinese folk medicine, is recognized for its anti-inflammatory and analgesic properties. In this study, we report the extraction, purification, and anti-inflammatory mechanisms of C-glycosides from A. decipiens (ADCG). We developed an integrated strategy for the efficient enrichment and identification of polar C-glycosides from A. decipiens. Sample pretreatment via freeze-drying preserved thermolabile constituents, and reflux extraction with 60% aqueous ethanol optimized polar compound recovery. A key innovation was the implementation of a repeated-loading strategy using polyamide resin chromatography, which significantly enhanced the dynamic adsorption capacity and prevented leakage of target analytes. This approach achieved a 15.8-fold enrichment of target C-glycosides and enabled the identification of seven compounds, including chrysin- and apigenin-based derivatives. It effectively provides an efficient and cost-effective approach for the large-scale enrichment of this fraction. ADCG significantly suppressed granuloma formation in a cotton ball-induced model. In vitro, ADCG demonstrated potent anti-inflammatory effects in LPS-stimulated RAW264.7 macrophages by dose-dependently inhibiting the production of NO, TNF-α, and IL-6, without inducing cytotoxicity. In TNF-α-induced human rheumatoid arthritis synovial MH7A cells, ADCG inhibited proliferation, migration, invasion, and the secretion of pro-inflammatory mediators IL-6, IL-8, IL-1β, and MMP-1. Mechanistic investigations combining molecular docking and Western blot analysis revealed that ADCG directly inhibits NF-κB activation by suppressing IκBα degradation and p65 phosphorylation, thereby blocking nuclear translocation and the expression of downstream pro-inflammatory genes. Furthermore, constituents of ADCG exhibited high binding affinity for JAK2 and EGFR, suggesting a potential multitarget mechanism. These findings underscore the potential of ADCG as a complementary therapeutic agent for inflammatory diseases, particularly rheumatoid arthritis, and provide important insights into its molecular mechanisms of action.