基于网络药理学、分子对接及实验验证探究杜仲抗骨质疏松症的作用机制OA

Objective To explore the core targets and mechanisms of Eucommia ulmoides in the treatment of osteoporosis through network pharmacology,molecular docking,and in vitro cell experiments.Methods Active components and targets of E.ulmoides were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),and osteoporosis-related disease targets were retrieved from GeneCards,DisGeNET,and OMIM databases.Protein-protein interaction network and"active component-common target"network were constructed.Key targets were analyzed for biological functions and signaling pathways using the Metascape database.Molecular docking between core components and key targets was performed and visualized using AutoDock software.An in vitro osteoblast model was established,and cell proliferation and mineralization were assessed by CCK-8 assay and Alizarin Red staining.Western blot was used to detect the protein expression of collagen I(Col-I),runt-related transcription factor 2(Runx2),and protein kinase B1(AKT1).Results A total of 28 active components of E.ulmoides,221 targets,and 2 661 osteoporosis-related disease targets were identified.Target mapping screened 80 potential targets for osteoporosis,with core targets including AKT1,tumor necrosis factor(TNF),and hypoxia-inducible factor 1α(HIF1A).Enrichment analysis identified six signaling pathways closely related to osteoporosis.Molecular docking showed that chlorogenic acid and quercetin from E.ulmoides can bind to AKT1,HIF1A,and TNF proteins.In vitro experiments demonstrated that chlorogenic acid,an active component of E.ulmoides,increased cell viability and upregulated the protein expression of AKT1,Col-I,and Runx2.Conclusion E.ulmoides may exert therapeutic effects in osteoporosis by regulating AKT1 expression and promoting osteoblast proliferation,differentiation,and mineralization.