In this study,the hypothesis that Wnt/β-catenin pathway is involved in the arterial calcification by regulating the osteoprotegerin(OPG)/receptor activator of NF-κB ligand(RANKL)system was tested.Theβ-catenin expre...In this study,the hypothesis that Wnt/β-catenin pathway is involved in the arterial calcification by regulating the osteoprotegerin(OPG)/receptor activator of NF-κB ligand(RANKL)system was tested.Theβ-catenin expression was measured in the warfarin-induced calcified arteries and the osteoblast-like cells differentiating from smooth muscle cells(SMCs)by immunohistochemistry and Western blotting.The Wnt/β-catenin pathway was activated or inhibited by lithium chloride(LiCl)or dickkopf 1(DKK1)in vitro and in vivo.Then the calcification level was determined by von Kossa staining,Ca^2+content assay,and alkaline phosphatase(ALP)activity assay.The expression levels of osteocalcin,OPG and RANKL were detected by Western blotting or real-time PCR.The results showed that in calcified arteries and OBL cells,the activation of Wnt/β-catenin pathway significantly enhanced the calcification as evidenced by increased von Kossa stains,Ca^2+contcnts,ALP activities,and osteocalcin expression levels(P<0.05),and it promoted the RANKL expression(P<0.05),but slightly affected the OPG expression.These results indicated that the activation of Wnt/β-catenin pathway worsens the arterial calcification,probably by promoting the RANKL expression.展开更多
文摘In this study,the hypothesis that Wnt/β-catenin pathway is involved in the arterial calcification by regulating the osteoprotegerin(OPG)/receptor activator of NF-κB ligand(RANKL)system was tested.Theβ-catenin expression was measured in the warfarin-induced calcified arteries and the osteoblast-like cells differentiating from smooth muscle cells(SMCs)by immunohistochemistry and Western blotting.The Wnt/β-catenin pathway was activated or inhibited by lithium chloride(LiCl)or dickkopf 1(DKK1)in vitro and in vivo.Then the calcification level was determined by von Kossa staining,Ca^2+content assay,and alkaline phosphatase(ALP)activity assay.The expression levels of osteocalcin,OPG and RANKL were detected by Western blotting or real-time PCR.The results showed that in calcified arteries and OBL cells,the activation of Wnt/β-catenin pathway significantly enhanced the calcification as evidenced by increased von Kossa stains,Ca^2+contcnts,ALP activities,and osteocalcin expression levels(P<0.05),and it promoted the RANKL expression(P<0.05),but slightly affected the OPG expression.These results indicated that the activation of Wnt/β-catenin pathway worsens the arterial calcification,probably by promoting the RANKL expression.
