Network pharmacology combined with molecular docking and molecular dynamics simulation to analyze the pharmacological mechanisms of Xueshuantong injection in the treatment of fracture

Background As a common clinical orthopedic injury, the healing process of fracture involves a complex molecular regulatory network. Xueshuantong injection (XI) containing (Ginsenoside Rb1, Ginsenoside Rg1, and Notoginsenoside R1), has a significant effect on promoting blood circulation, removing blood stasis, and improving microcirculation. However, the pharmacological mechanisms of XI in the treatment of fractures are not clear. Methods This study employed network pharmacology to screen for common targets between XI and fractures, and further analyzed to obtain core targets via protein-protein interaction (PPI) network. Subsequently, the expression characteristics of the targets were analyzed using the GSE17825 dataset. Molecular docking and molecular dynamics simulations were conducted to verify the binding capacity and stability of the active ingredients with the core targets. The mechanism of action was elucidated through MC3T3-E1 cell experiments and Akt1 knockdown experiments. Results Five key targets were identified, including AKT1, CASP3, IL1B, IL6, and STAT3. The three active ingredients had strong binding ability to Akt1 (the binding energy ranging from −7.7 to −8.0 kcal/mol) and stable binding. XI treatment significantly enhanced ALP activity, RUNX2 mRNA, and OCN protein expression, increased the number of calcified nodules, inhibited caspase 3 activity, and upregulated Akt1 expression in MC3T3 cells after osteogenic induction. Knockdown of Akt1 reversed the promotive effect of osteogenic differentiation. Conclusion XI synergistically binds to Akt1 through its active ingredients, thereby upregulating its expression, promoting osteoblast differentiation, inhibiting apoptosis, and accelerating fracture healing. This study systematically elucidates the molecular mechanism of its treatment of fractures, and provides a scientific basis for clinical application and related drug development.