Screening of Anti-inflammatory Active Components of Lepyrodiclis holosteoides Based on LC-MS and Molecular Docking Technology

Objective: To screen the anti-inflammatory active constituents in Lepyrodiclis holosteoides based on high performance liquid chromatography-mass spectrometry (HPLC-MS) and molecular docking technology. Methods: In order to explore the anti-inflammatory active fraction of Lepyrodiclis holosteoides, inhibitory effects of different polar fractions on nitric oxide (NO), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) production in lipopolysaccharide (LPS) stimulated RAW 264.7 were studied. The chemical components of active site were analyzed by HPLC-MS. Five important inflammatory protein receptors such as TNF-α, IL-6, interleukin-1β (IL-1β), prostaglandin E₂ (PGE₂) and nuclear transcription factor-κB (NF-κB) were selected, and the active ingredients were molecularly docked through Auto Dock Vina software. Results: Experiments in vitro showed that the ethyl acetate fraction could inhibit the production of NO, TNF-α and IL-6 in LPS induced RAW 264.7 cells. Compared with the model control group, inhibitory rates of NO, TNF-α and IL-6 were significantly lower at the highest concentration of 75 μg/mL (P<0.01), which were 63.53%, 34.23% and 34.58% respectively. Eleven chemical components were identified by HPLC-MS, including 8 flavonoids (rutin, vitexin, kaempferol, rhamnazin, quercetin, apigenin, apigenin-6-C-glucose-8-C-xyloside, quercetin) and 3 coumarins (umbelliferone, 7-methoxy coumarin, 5,7-dihydroxy coumarin). Molecular docking results showed that 5 core inflammatory targets had the strongest binding affinity to quercetin, kaempferol, rutin, quercetin and vitexin, with a binding energy of −8.5, −7.8, −8.0, −7.2 and −10.0 kcal/mol respectively, which were superior to standard drug dexamethasone. The results indicated that the ingredients could site bind to 5 target proteins. Conclusion: The ethyl acetate fraction of Lepyrodiclis holosteoides had anti-inflammatory activity. This study clarified the potential material basis of Lepyrodiclis holosteoides quickly and conveniently by using HPLC-MS combined with molecular docking technology.