Department of Geriatrics, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430014, China.
Department of Geriatrics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
Curr Med Sci. 2019 Feb;39(1):28-36. doi: 10.1007/s11596-019-1996-4. Epub 2019 Mar 13.
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. The β-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 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 contents, 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.
在这项研究中，我们检测了 Wnt/β-连环蛋白通路通过调节护骨素（OPG）/核因子-κB 受体激活剂配体（RANKL）系统参与动脉钙化的假说。通过免疫组织化学和 Western blot 法测量了华法林诱导的钙化动脉和由平滑肌细胞（SMCs）分化而来的成骨样细胞中的β-连环蛋白表达。通过氯化锂（LiCl）或 Dickkopf 1（DKK1）在体外和体内激活或抑制 Wnt/β-连环蛋白通路。然后通过 von Kossa 染色、Ca 含量测定和碱性磷酸酶（ALP）活性测定来确定钙化水平。通过 Western blot 或实时 PCR 检测骨钙素、OPG 和 RANKL 的表达水平。结果表明，在钙化动脉和 OBL 细胞中，Wnt/β-连环蛋白通路的激活显著增强了钙化，表现为 von Kossa 染色、Ca 含量、ALP 活性和骨钙素表达水平增加（P<0.05），并促进了 RANKL 表达（P<0.05），但对 OPG 表达影响较小。这些结果表明，Wnt/β-连环蛋白通路的激活使动脉钙化恶化，可能是通过促进 RANKL 表达。
