Background:Gushukang (GSK), a traditional Chinese medical prescription, has
made a great and extensive contribution to the treatment of different forms of osteoporosis, but
polypharmacology studies of its mechanism of action are lacking. This study investigates
the pharmacological mechanism of osteoporosis using network pharmacology and molecular
docking. Experimental verification was carried out to confirm the efficacy of GSK on RANKLinduced
osteoclast differentiation in RAW264.7 cells to verify the network pharmacology
studies.Methods:The effective chemical components and corresponding targets of osteoporosis
with oral bioavailability of more than 30% and drug-like properties greater than 0.18 were
searched in the TCMSP and TCM-ID databases. DrugBank, GeneCards, OMIM, TTD, and
other databases were examined for targets related to osteoporosis. Using Cytoscape software,
a network of possible TCM-active ingredient-osteoporosis targets was created. STRING
software was used to create the networks of protein-protein interactions. The DAVID
program was carried out to conduct GO and KEGG pathway enrichment analyses of the
targets. Molecular docking and pattern of action analysis were carried out using software
like AutoDock Vina and Discovery Studio Visualizer. The growth media for RAW264.7
cells contained varying doses of GSK serum and 50 ng/mL RANKL. The activity of TRAP
was altered. Additionally, genes related to osteoclasts were examined using an RT-PCR
assay.Results:Network pharmacological analysis revealed that the primary efficacy targets of osteoporosis
were PTGS2, PTGS1, HSP90AA1, NCOA2, ADRB2, ESR1, NCOA1, and AR. The
pharmacological targets of osteoporosis may be mediated by substances including quercetin,
kaempferol, luteolin, naringenin, icariin, anthocyanin, tanshinone IIA, and cryptotanshinone.
GSK markedly inhibited RANKL-induced TRAP activity. qRT-PCR results revealed decreased
expression of the PTGS2 and ADRB2 genes upon GSK treatment.Conclusion:The findings of network pharmacology, molecular docking, as well as experimental
verification provide a new further study for elucidating the pharmacodynamic substance
basis and polypharmacology mechanism of GSK in treating osteoporosis.