Active component screening of ginseng for postmenopausal osteoporosis using ESR2 molecular docking and...

Ginseng exhibits pharmacological efficacy in the treatment of female postmenopausal osteoporosis (PMOP), although its specific active constituents remain unidentified. This study aims to identify the anti-PMOP active components of ginseng by screening against the ESR2 enzyme. Using osteoprotegerin (OPG) as a biomarker, the most effective ESR2-active components of ginseng were determined through experiments with MC3T3-E1 cells. The potential active components within ginseng were analyzed using ultra-high-performance liquid chromatography coupled with quadrupole-exactive orbitrap mass spectrometry (UHPLC-QE Orbitrap-MS) and further validated through molecular docking and molecular dynamics simulations. The single-target affinity ultrafiltration method was employed to assess the affinity rate (AR%) of the ESR2 enzyme by analyzing peak areas, leading to the identification of key active components of ginseng effective against PMOP through molecular docking analysis. Experimental findings indicated that ginseng extract exhibits compatibility with the ESR2 enzyme, with the 95 % ethanol extract demonstrating the highest activity. A total of 35 potential active components were identified through UHPLC-QE Orbitrap-MS analysis. Following a comprehensive evaluation of molecular docking binding energy and AR% results, Ginsenoside Rf, Ginsenoside Re, and Ginsenoside Rb1 were confirmed as the principal active components of ginseng effective against PMOP. The survival rate, OPG content, and alkaline phosphatase (ALP) activity index, a marker of osteogenic differentiation, were utilized to confirm the final key active components. In this study, the ESR2 enzyme was identified as the active target, and a combination of cell experiments, molecular docking, kinetic simulations, and affinity ultrafiltration was employed to effectively screen the active anti-PMOP constituents of ginseng.