The Research On The Effect And Mechanism Of Taxifolin In Osteoclastogenesis And Function Both In Vitro And In Vivo

Objective(1)To clarify the effects of taxifolin on osteoclastogenesis and osteoclast function in vitro.(2)To investigate the effects of taxifolin on bone loss and other aspects in ovariectomized mice in vivo.Methods(1)In vitro experiments: RANKL was used to induce the osteoclast differentiation of cell line RAW 264.7 and mouse primary bone marrow mononuclear cells(adding macrophage colony forming factor),and different concentrations of taxifolin were added for intervention.The effects of taxifolin on osteoclast formation and function were investigated through TRAP staining,TRAP enzyme activity assay,bone resorption assay,cytoskeletal fibrillar actin loop staining,osteoclast-specific gene,and protein detection.(2)In vivo experiments: The animal model of osteoporosis was established by resecting the ovaries of C57/BL6 female mice.The experimental group was given taxifolin intraperitoneally,while the control group was given the same dose of saline.At the endpoint of the experiment,the morphological and histological evaluation of bone tissue in each group were performed via section staining and μ-CT scanning.The levels of inflammatory factors in peripheral blood of each group were detected by enzyme-linked immunosorbent assay,in order to investigate the effects of taxifolin on bone loss and other aspects in ovariectomized mice in vivo.Results It was observed in vitro that taxifolin could inhibit osteoclast formation and bone resorption function,as well as the activation of nuclear factor-κB,C-Fos,and MAPK.Expression of osteoclast-specific genes,including TRAP,MMP-9,Cathepsin K,C-Fos,NFATc1 and RANK was also reduced.Inhibition of nuclear transcription of transcription factors NF-κB and AP-1,and prevention the production of reactive oxygen species were both detected.The mechanism of aforementioned effects might be the inhibition of NF-κB,MAPKs and AKT protein pathways.In addition,we found that taxifolin could inhibit the osteoclast activity and reduce the serum levels of TNF-α、IL-1β、IL-6 and RANKL in vivo,as well as the bone loss induced by ovariectomy.Conclusion(1)Taxifolin inhibited osteoclastogenesis and osteoclast function,and alleviated bone loss caused by ovariectomy in mice.(2)Taxifolin may be a new strategy for the treatment of osteoclast-related diseases.