Hydatid Antigen B Promote RANKL/NF-κB/TAK1-mediated Oste-oclastogenesis via Inhibition of TAZ
Objective To investigate the molecular mechanisms of hydatid antigen-B(Hyd-B)in osteoclastogenesis.Methods Cell experiment group-1:Bone marrow mesenchymal stem cells(BMSCs)were divided into 3 groups(Control group,MCSF+RANKL group,MCSF+RANKL+Hyd-B group),cells in the MCSF+RANKL group and MCSF+RANKL+Hyd-B group were in-duced to differentiate into osteoclasts by using macrophage colony-stimulating factor(MCSF)com-bined with soluble receptor activator of nuclear factor-Kb ligand(RANKL),and then treated with or without Hyd-B respectively.Cell experiment group-2:BMSCs were divided into Ctrl group and Hyd-B treatment group(Treat group),the effect of Hyd-B on the direct interaction between TAZ and TAK1 was determined by co-immunoprecipitation(co-IP).IP was performed by using anti-TAK1 antibody,and TAZ expression level was detected by IB.Cell experiment group-3:BMSCs were divided into 5 groups:Control group,MCSF+RANKL group,MCSF+RANKL+Hyd-B group,MCSF+RANKL+Hyd-B+TAZ-OE group and MCSF+RANKL+Hyd-B+OE-vector group.TAZ overexpression plasmid or vector plasmid was transfected to the BMSCs in the MCSF+RANKL+Hyd-B+TAZ-OE group and MCSF+RANKL+Hyd-B+OE-vector group respective-ly.qPCR was used to detect the mRNA expression levels of osteoclast differentiation markers of TRAP and Cathepsin K.Western blotting was used to detect the expression levels of nuclear p-P65,cytoplasmic P65,nuclear NFATc1,p-AKT,AKT,p-ERK 1/2,ERK 1/2,p-TAZ,TAZ and TAK1.Results Compared with those in the Control group,the TRAP and Cathepsin K mRNA ex-pression levels in the MCSF+RANKL and MCSF+RANKL+Hyd-B groups were increased(P＜0.05);the expression levels of p-P65,p-AKT and p-ERK 1/2 were increased(P＜0.05);the expression levels of p-P65 and NFATc1 in nucleus were increased(P＜0.05).The expression lev-els of the above indicators were further increased in the MCSF+RANKL+Hyd-B group when com-pared with those in the MCSF+RANKL group(P＜0.05).Compared with those in the MCSF+RANKL+Hyd-B group,the expression levels of the above indicators in MCSF+RANKL+Hyd-B+TAZ-OE group were all reduced(P＜0.05).Co-IP results showed that Hyd-B treatment enhanced the interaction between TAZ and TAK1(P＜0.05).Compared with those in the Ctrl group,the phosphorylated level of cytoplasmic TAZ in Treat group was increased,the expression level of TAZ was reduced(P＜0.05).Conclusion Hyd-B can promot the RANKL/NF-κB/TAK1-mediated os-teoclastogenesis through inhibition of TAZ.
bone cystic echinococcosishydatid antigen BRANKL/NF-κB/TAK1tran-scriptional coactivator with PDZ-binding motifosteoclastogenesis
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