The mechanism of action and experimental Verification of Narenmandula in the treatment of postmenopausal osteoporosis

Background: Narenmandula is a classic ancient remedy in Inner Mongolia, historically used for gastrointestinal diseases. In recent decades, Inner Mongolia Medical University found that it has a significant effect in promoting fracture healing and increasing bone density, and has been used to treat postmenopausal osteoporosis (PMOP), but its mechanism is unclear. Objective: Identify the mechanism of action of Narenmandula for PMOP treatment. Methods: Network pharmacology, molecular docking and ovarian departing rat models were used to verify the relevant mechanism of Narenmandula in the treatment of PMOP. Results: We confirmed that NRMDL prescription can improve OVX-induced bone loss, improve trabecular density, and relieve osteoporosis. Upon screening of network pharmacology, we ob-tained 238 overlapping genes of Narenmandula and PMOP, and analyzed AKT, IL1B, and IL6 as key genes by network topology. Among the 1143 target genes that interact with PMOP, 107 NRMDL active compounds correspond to 345 target genes and 238 overlapping genes. Network topology analysis showed the top 8 active ingredients, such as quercetin and kaempferol, and the top 20 key genes, such as AKT, IL1B, IL6, INS, JUN, STAT3, TNF, TP53, etc. Enrichment analysis revealed involvement of PI3K-Akt, HIF-1, FoxO, MAPK, and TNF signaling pathways. In addition, we found the most important active compounds bind tightly to core proteins, which were verified by molecular docking analysis. The AKT-related pathway had good binding energy, and the pathway was verified by cell and animal experiments. Conclusion: The potential mechanism and efficacy of Narenmandula against PMOP may be related to the PI3K-AKT pathway.