基于网络药理学及体外实验探究黄精筋骨方治疗老年性骨质疏松症的作用机制OA

Objective:To investigate the therapeutic mechanism of Huang-Jing-Jin-Gu-Fang(HJJGF)in senile osteoporosis based on network pharmacology and to validate its effects through cellular experiments.Methods:Network pharmacology was used to analyze the potential active components,core targets,and related signaling pathways of HJJGF in the treat-ment of senile osteoporosis.Molecular docking was performed to verify the binding activities between key active com-pounds and core targets.An H2O2-induced senescent bone marrow mesenchymal stem cell(BMSC)model was estab-lished to evaluate the effects of HJJGF on cellular senescence,osteogenic/adipogenic differentiation,mitochondrial func-tion,and lipid droplet-mitochondria interactions.The expression levels of AMPK-Mfn2-related proteins were also detec-ted.Results:A total of 182 potential targets of HJJGF for the treatment of senile osteoporosis were identified through network pharmacology.The core targets included AKT1,PPARG,and IGF1,and were mainly enriched in signaling pathways such as AMPK and PI3K-Akt.Molecular docking results showed that the key active components quercetin,kaempferol,naringenin,and nobiletin exhibited good binding activity with key proteins of the AMPK pathway,including AKT1,PPARG,and IGF1.In vitro experiments demonstra-ted that HJJGF significantly reduced BMSC senescence,promoted osteogenic differentiation,and inhibited adipogenic dif-ferentiation.Meanwhile,HJJGF upregulated the protein expression levels of p-AMPK/AMPK and Mfn2 in senescent BMSCs,enhanced lipid droplet-mitochondria interactions,improved mitochondrial energy metabolism,increased the NAD+/NADH ratio,and reduced ROS accumulation.Conclusion:HJJGF may exert anti-senile osteoporosis effects by reg-ulating the AMPK-Mfn2 signaling axis,improving lipid droplet-mitochondria interactions and mitochondrial function,thereby delaying BMSC senescence.