Integrated Network Pharmacology and Molecular Dynamics Simulation to Evaluate the Anti-Osteoporotic Efficacy of Radish (Raphanus sativus) Seeds

Abstract

Glucocorticoid-induced osteoporosis (GIO), a prevalent secondary bone disorder driven by suppressed bone
formation, heightened resorption, and oxidative stress, prompts the exploration of natural alternatives to
conventional treatments burdened by side effects; thus, this study investigates the anti-osteoporotic
potential and molecular mechanisms of Raphanus sativus seed extract against dexamethasone-induced bone
loss via an integrated computational and experimental approach. Bioactive compounds from R. sativus
seeds were sourced from literature and databases, with network pharmacology revealing 33
phytocompounds and 219 overlapping targets with GIO; hub proteins such as TNF, STAT3, NFKB1, and PIK3R1
emerged from PPI networks, linked to inflammatory signaling and bone remodeling, while KEGG enrichment
underscored PI3K-Akt and osteoclast differentiation pathways. Molecular docking and dynamics (MD)
simulations confirmed stable binding of lead compounds-Castasterone to NFKB1 and S-methyl-L-cysteine
sulfoxide to PIK3R1-while in vitro assays demonstrated potent antioxidant activity (DPPH IC₅₀: 58.24 μg/mL;
NO IC₅₀: 62.22 μg/mL) alongside high total phenolic content (TPC: 76.30 mg GAE/g) and flavonoid content
(TFC: 44.7 mg QE/g). These findings elucidate how R. sativus seeds protect against glucocorticoid-induced
bone damage by modulating inflammation, countering oxidative stress, and restoring bone homeostasis,
laying a mechanistic groundwork for its development as a natural therapeutic for osteoporosis management.