ABSTRACT
Aim To investigate the effect and mechanism of Eucommiae ulmoides and Radix Dipsaci in the treatment of osteoporosis.Methods The active ingredients of Eucommiae ulmoides and Radix Dipsaci were obtained by TCMSP,BATMAN-TCM platform and the literature.Osteoporosis related genes were collected by GeneCards and OMIM.The compound target network was constructed with Cytoscape 3.7.2 software.The protein interaction network was constructed with STRING online website.DAVID was used to perform gene ontology(GO)enrichment analysis and kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.The synergistic effect of Eucommiae ulmoides and Radix Dipsaci was tested by osteo-blast proliferation,alkaline phosphatase expression and mineralized nodule formation experiments.The protein expressions of BMP2 and Wnt signaling pathway were investigated by Western blotting.Results A total of 44 active components of Eucommiae ulmoides and 17 active components of Radix Dipsaci were screened,and 80 targets were intersected by osteoporosis.GO analysis showed that the action mechanism of Eucommiae ulmoides and Radix Dipsaci in the treatment of osteoporosis was involved with 52 items of biological process,8 items of cell composition and 16 items of molecular function.The KEGG enrichment pathway was dominated by 6 major signaling pathways.Compared with the control group,the osteoblast proliferation,alkaline phosphatase expression and mineralized nodule formation significantly increased in Eucommiae ulmoides or Radix Dipsaci group(P<0.01).Western blotting showed that Eucommiae ulmoides mainly regulated the expression levels of Wnt signaling pathways(P<0.05 or P<0.01),and Radix Dipsaci up-regulated the expression levels of BMP2 signaling pathways(P<0.01).Meanwhile,Eucommiae ulmoides and Radix Dipsaci significantly enhanced these effects,respectively(P<0.01).Conclusions Although Eucommiae ulmoides and Radix Dipsaci have different active components,their main targets and pathways are the same in the treatment of osteoporosis.Eucommiae ulmoides and Radix Dipsaci can regulate the function of osteoblasts through BMP2 and Wnt signaling pathways,and the combination of the two drugs in the treatment of osteoporosis has synergistic effect.