Vascular Calcification in Chronic Kidney Disease is Induced by Bone Morphogenetic Protein-2 via a Mechanism Involving the Wnt/β-Catenin Pathway
Vascular Calcification in Chronic Kidney Disease is Induced by Bone Morphogenetic Protein-2 via a Mechanism Involving the Wnt/β-Catenin Pathway
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DOI：
10.1159/000366400
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发表时间：
2014-01-01
影响因子：
--
通讯作者：
Yuan, Weijie
中科院分区：
文献类型：
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作者：
Rong, Shu;Zhao, Xuezhi;Yuan, Weijie
背景资料：血管钙化（VC）是慢性肾脏病（CKD）动脉粥样硬化加速过程的危险因素之一，其中血管平滑肌细胞（VSMC）经历成骨样细胞的表型转化。磷酸盐是VC的重要调节剂。研究方法：通过qRT-PCR和蛋白质印迹法检测大鼠VSMCs中不同平滑肌细胞或成骨标志物对高浓度磷酸盐或外源性骨形态发生蛋白2（BMP-2）的表达。放射免疫法测定骨钙素分泌。碱性磷酸酶（ALP）活性测定和茜素染色检测VSMCs的分化和钙化。进行短发夹RNA介导的β-连环蛋白沉默以检查Wnt/β-连环蛋白信号传导在高磷酸盐或BMP-2诱导的VSMC钙化和成骨细胞分化中的参与。TUNEL法和免疫荧光法检测细胞凋亡。结果：CKD患者血清BMP-2水平显著高于对照组。高磷酸盐浓度和BMP-2诱导VSMC凋亡，并上调β-连环蛋白，Msx 2，Runx 2和磷酸盐协同转运蛋白Pitl的表达，而BMP-2中和抗体逆转了这些作用。敲低β-连环蛋白可消除高磷酸盐和BMP-2对VSMC凋亡和钙化的影响。结论：BMP-2通过Wnt/β-catenin通路在CKD患者VSMCs和VC钙沉积中发挥重要作用。版权所有（C）2014 S. Karger AG，巴塞尔
Background: Vascular calcification (VC), in which vascular smooth muscle cells (VSMCs) undergo a phenotypic transformation into osteoblast-like cells, is one of the emergent risk factors for the accelerated atherosclerosis process characteristic of chronic kidney disease (CKD). Phosphate is an important regulator of VC. Methods: The expression of different smooth muscle cell or osteogenesis markers in response to high concentrations of phosphate or exogenous bone morphogenetic protein 2 (BMP-2) was examined by qRT-PCR and western blotting in rat VSMCs. Osteocalcin secretion was measured by radioimmunoassay. Differentiation and calcification of VSMCs were examined by alkaline phosphatase (ALP) activity assay and Alizarin staining. Short hairpin RNA-mediated silencing of p-catenin was performed to examine the involvement of Wnt/beta-catenin signaling in VSMC calcification and osteoblastic differentiation induced by high phosphate or BMP-2. Apoptosis was determined by TUNEL assay and immunofluorescence imaging. Results: BMP-2 serum levels were significantly higher in CKD patients than in controls. High phosphate concentrations and BMP-2 induced VSMC apoptosis and upregulated the expression of beta-catenin, Msx2, Runx2 and the phosphate cotransporter Pitl, whereas a BMP-2 neutralization antibody reversed these effects. Knockdown of beta-catenin abolished the effect of high phosphate and BMP-2 on VSMC apoptosis and calcification. Conclusions: BMP-2 plays a crucial role in calcium deposition in VSMCs and VC in CKD patients via a mechanism involving the Wnt/beta-catenin pathway. Copyright (C) 2014 S. Karger AG, Basel