Potential Mechanism of Gumibao Decoction in Treating Glucocorticoidinduced Osteoporosis Based on Network Pharmacology...

Gumibao decoction, a traditional Chinese herbal prescription, has demonstrated promising effects in treating osteoporosis; however, the underlying mechanism remains unclear. We investigated the regulatory mechanisms of Gumibao decoction in osteoporosis through network pharmacology analysis and validated its therapeutic effects using animal experiments.

The TCMSP database was used to screen the bioactive constituents of Gumibao decoction and identify their associated targets. Disease targets for osteoporosis were acquired through the Gene- Cards, PharmGKB, DrugBank, OMIM, and TTD databases. A Protein-Protein Interaction (PPI) network was generated. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were subsequently performed for the core targets identified in the PPI network. Moreover, a glucocorticoid-induced osteoporosis model was established in Sprague-Dawley (SD) rats by administering methylprednisolone and dexamethasone, and the regulatory effects of Gumibao decoction on osteoporosis were evaluated using bone mass detection and western blotting.

The network pharmacology analysis identified quercetin, (-)-Epigallocatechin-3-Gallate (EGCG), kaempferol, luteolin, and fisetin as the primary bioactive components of Gumibao decoction. Core target proteins included AKT1, TP53, JUN, CTNNB1, and IL1B. The KEGG pathway enrichment analysis revealed the significant involvement of the TGF-β signalling pathway, osteoclast differentiation, and the MAPK signalling pathway in mediating its anti-osteoporotic effects. In vivo validation demonstrated that Gumibao decoction significantly ameliorated glucocorticoid-induced reductions in Bone Mineral Density (BMD) and deterioration of bone microstructure. Furthermore, protein expression analysis revealed significantly reduced levels of Smad4, phospho-Smad2/3, and TGF-β1 in the model group compared with the blank control group. Notably, all Gumibao decoction treatment groups exhibited significant upregulation of Smad4, P-Smad2/3, and TGF-β1 expression compared with the model group, validating the network pharmacology predictions implicating the TGF-β pathway.

Research on the components of Gumibao decoction has shown that it can regulate homeostasis between osteoblasts and osteoclasts through multiple targeted pathways, thereby positively modulating bone density, bone microstructure, and bone markers. This ultimately inhibits osteoclast differentiation and stimulates osteoblast proliferation, effectively alleviating and preventing osteoporosis.

Gumibao decoction effectively improves glucocorticoid-induced osteoporosis, potentially by upregulating the expression of TGF-β1, P-Smad2/3, and Smad4 through the TGF-Β/Smad pathway, promoting bone formation, and restoring bone metabolic balance in glucocorticoid-induced osteoporosis rats.