Capture the superior anti-osteoporotic innovative multi-component complex from Ligustri Lucidi Fructus based on...

Postmenopausal osteoporosis (PMOP) was accompanied by iron overload and low levels of calcium (Ca), phosphorus (Pi) and magnesium (Mg), which required downregulating iron and upregulating Ca/Pi/Mg simultaneously. It was confirmed that Ligustri Lucidi Fructus (FLL) had the ability to repair above minerals metabolism bidirectionally, but its pharmacological substance and mechanism were obscure. Previous research revealed the different ratio of Phenylethanols, Iridoids and Triterpenoids in FLL lead to its efficacy changes in anti-PMOP.

To capture the superior anti-osteoporotic mixture from raw FLL based on Component Recombination strategy, and explore its potential mechanisms from the perspective of bidirectionally repair mineral metabolism.

The small molecule compounds (Phenylethanols, iridoids, triterpenes) and polysaccharide were determined by HPLC-DAD and sulfuric acid-anthrone method respectively. The zebrafish model was not only used to study the pharmacological interactions between multiple components, but also for Orthogonal experimental screening of the superior composition ratio among multiple components. Subsequently, PMOP rat model combined with transcriptomics research was carried out to disclose the potential mechanism of FLL and obtained composition (FLLC) in bidirectional repair of mineral metabolism and treating PMOP. Molecular docking analysis was used to predict the molecular docking, calculate binding energies, visualize proteins and small molecule ligands. Eventually, the PMOP rat model was applied again to systematically compare the efficacy between optimized out FLLC and FLL according to bone mineral density (BMD), Micro-CT analysis, mineral metabolism indicators, bone metabolism indicators and Western Blot (WB) analysis.

When 95 % and 70 % ethanol extracts at a mass ratio 1: 12 (FLLC), which possessed the optimal ability to resist PMOP. The combinatorial index (CI) analysis indicated that multi-components in FLL synergistically promoted bone mineralization (CI <1). RNA sequencing combined with molecular docking revealed that PTH1R/PKA pathway was one of the related mechanisms of FLL in bidirectionally repair mineral metabolism. Rats model verified that FLLC and FLL increased the expression of PTH1R, cAMP, p-PKA, BMP2 and reduced the expression of SOST, FLLC was more superior than FLL (p < 0.05). Both of FLL and FLLC significantly improved BMD and bone microstructure, repaired bone metabolism balance, reversed mineral metabolism disorder. Moreover, FLLC significantly strengthened the beneficial effect in anti-PMOP compared to FLL (p < 0.05).

The optimized FLLC bidirectionally reversed mineral metabolism disorders through PTH1R/PKA pathway, and its enhanced anti-PMOP activity was achieved by components recombination. The component recombination strategy continuously boosted the pharmacodynamics of natural herbs, originating from nature but surpassing nature.