当归补血汤调控TNF-α/PI3K-AKT/BMP信号通路改善糖尿病性骨质疏松的机制研究OA

Objective:To investigate the potential targets and mechanisms of action of Danggui Buxue Decoction(DGBXD)for the treatment of diabetic osteoporosis(DOP)based on network pharmacology,molecular docking,and animal experiments.Methods:The active chemical components of DGBXD were screened by a network pharmacology approach to retrieve the targets of DGBXD related to diabetes osteoporosis.Protein-protein interaction(PPI)networks of drug-disease targets were constructed.Validate the interactions between major targets and active chemical components by molecular docking.Using a db/db diabetic mouse model,the potential targets of DGBXD intervention in DOP were validated and the mechanism of action was explored by ELISA,Western blot and pathology.Results:A total of 413 targets for the action of Astragalus and 65 targets for the action of Angelica sinensis were obtained by network pharmacology,and 15 330 genes were retrieved as DOP-related disease targets.The total number of targets of DGBXD for DOP was 202,including 190 from Astragalus and 11 from Angelica sinensis.Network phar-macological enrichment analysis indicated that DGBXD may improve osteogenic abnormalities in DOP by regulating inflammatory factor expression through modulating TNF-α expression and PI3K-AKT signaling pathway.The results of animal experiments showed that compared to the control group,the serum IL-6,IL-8 and TNF-α levels were significantly increased in the model group(P<0.05);the intervention of DGBXD could inhibit the excessive IL-6(P<0.05),IL-8(P<0.05)and TNF-α(P<0.05)expressions in the model group.Pathological results showed that DGBXD could correct the disorders of bone formation caused by abnormal inflammatory response.Western blot results further indicated that compared to the model group,the interven-tion of DGBXD could significantly reduce the protein expression of TNF-α,AKT,phosphorylated AKT,and correct the abnor-mal protein expression of BMP2(P<0.05).Conclusion:DGBXD may inhibit the inflammatory response and enhance the expres-sion of BMP2 by regulating the TNF-α/PI3K-AKT/BMP signaling pathway,thus promoting the synthesis and mineralization of bone matrix,alleviating the high glucose-induced osteogenic inhibition,improving the balance of bone metabolism,and achieving the therapeutic effect on DOP.