Selection of the Anti-Osteoporosis Active Ingredients of Fructus Psoraleae-Eucommia-Drynariae Rhizoma Based on...

Osteoporosis (OP) is a prevalent metabolic bone disease globally. Currently, the development of Traditional Chinese Medicine (TCM) resources to unblock joints, strengthen bones, and enhance muscle function to regulate anti-osteogenic and anabolic metabolism and thus reshape intraosseous homeostasis was an effective way to alleviate OP. The F-E-D formula, comprising Fructus Psoraleae, Eucommia, and Drynariae Rhizoma, has shown efficacy in treating OP. However, its complex natural components necessitate the screening and simplification of bioactive compounds to further elucidate their therapeutic mechanisms and enhance therapeutic efficacy. In this study, we first used drug-target binding to produce different effects, which in turn exhibited different retention characteristics on the stationary phase. Using osteoblasts and osteoclasts as stationary phases, a chromatographic system (Solid-phase Bio-cell Chromatography, SBC) had been constructed to mimic the drug-target interaction, and the separation, analysis, and bioactivity screening of the chemical components of F-E-D had been performed. Then, the above collected eluates were analyzed by fine metabolomics, and 95 effective metabolites were initially screened and combined with database screening to finally select betaine, L-fucose, and itaconic acid as potentially active candidate compound monomers for the interaction with osteoblast-osteoclast in F-E-D. In terms of cell validation experiments, we found that the screened active monomers significantly inhibited the formation of osteoclasts, and the itaconic acid-treated group played a significant inhibitory effect on the expression of inflammatory factors TNF-α and IL-6. The above experimental data showed that the monomeric active ingredients in TCM could be effectively screened by solid-phase bio-chromatography and HPLC-MS, and the in vitro cellular experiments verified that the active monomers of TCM slowed down the progression of OP by inhibiting osteoclast production and alleviating the expression of inflammation.