Identification of Anti‐Inflammatory Active Components in Morinda officinalis via Spectroscopic Effect Relationship Analysis and NF‐κB Pathway Inhibition

Morindae Officinalis Radix (MOR), the dried root of Morinda officinalis How, contains abundant saccharides and is used clinically after different processing procedures. However, the anti-inflammatory material basis of MOR and the chemical consequences of Lycium barbarum processing remain insufficiently defined. In this study, high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) was developed for fingerprint analysis of 15 batches of raw MOR and the corresponding L. barbarum-processed MOR (LB-MOR). Seventeen common peaks were resolved, and 11 saccharides were assigned by comparison with reference standards. Anti-inflammatory activity was evaluated in LPS-stimulated RAW 264.7 macrophages by quantifying nitric oxide (NO), TNF-α, IL-1β, and IL-6. Grey relational analysis and partial least squares regression linked the HPLC fingerprints with bioactivity and identified glucose, sucrose, kestose, and nystose as the principal activity-associated saccharides. Subsequent validation showed that these saccharides inhibited inflammatory mediator release, with kestose and nystose showing the strongest effects. Western blotting further indicated that kestose and nystose restored IκBα expression and decreased COX-2 expression and the p-p65/p65 ratio, consistent with suppression of sustained NF-κB signaling. This separation-based fingerprint and spectrum-effect workflow identify anti-inflammatory oligosaccharides in MOR and provides candidate quality markers for raw and processed MOR products.