Fibronectin enhances in vitro vascular calcification by promoting osteoblastic differentiation of vascular smooth muscle cells via ERK pathway
Fibronectin enhances in vitro vascular calcification by promoting osteoblastic differentiation of vascular smooth muscle cells via ERK pathway
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DOI：
10.1002/jcb.20999
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发表时间：
2006-12-01
影响因子：
4
通讯作者：
Wang, Kui
中科院分区：
文献类型：
--
作者：
Bing, Hai-Tao;Wang, Chen-Guang;Wang, Kui
血管钙化的过程呈现出与成骨细胞分化相似的特征，其中纤维连接蛋白（FN）作为成骨细胞分化的调节因子，ERK信号通路参与其中。为了探讨FN是否通过ERK信号通路促进血管平滑肌细胞（VSMCs）向成骨细胞分化，我们采用体外细胞模型研究了FN对VSMCs钙化的影响。用10 mM β-甘油磷酸钠（β-GP）诱导在FN（0-20 pg/cm（2））涂层平板中培养的VSMC钙化。FN以剂量和时间依赖性方式加重VSMC钙化，如每张载玻片上钙化结节的数量和沉积物中的钙含量所示。RT-PCR和免疫印迹分析结果显示，FN还增强了成骨细胞的几种表型标志物的表达，包括碱性磷酸酶（ALP）活性、骨钙素（OC）和成骨细胞分化中的关键转录因子Osf 2/Cbfa 1。此外，ERK的特异性抑制剂PD 98059（10 μ M），显着抑制FN对钙化和表型标记物表达的影响。FN通过ERK信号通路促进VSMCs向成骨细胞分化，从而促进血管钙化。
The process of vascular calcification presents several features similar to osteogenesis in which fibronectin (FN) acts as a regulator of osteoblastic differentiation and the ERK signal pathway is involved. In order to find whether FN promotes the osteoblastic differentiation of vascular smooth muscle cells (VSMCs) through the ERK signal pathway, we investigated the effect of FN on the calcification of VSMCs by using an in vitro cell model. VSMCs cultured in plates with FN (0-20 pg/cm(2)) coating were induced to calcify by 10 mM sodium beta-glycerophosphate (beta-GP). FN exacerbated VSMC calcification in a dose- and time-dependent manner, as indicated by the number of calcifying nodules per slide and the amount of calcium in, the deposition. Data from RT-PCR and immunoblotting assay revealed that FN also enhanced the expression of several phenotypic markers of osteoblasts, including alkaline phosphatase (ALP) activity, osteocalcin (OC), and Osf2/Cbfa1, a key transcription factor in osteoblastic differentiation. Furthermore, a specific inhibitor for ERK, PD98059 (10 mu M), significantly suppressed the effect of FN on calcification and phenotypic marker expression. These findings seem to suggest that FN enhanced vascular calcification by promoting the osteoblastic differentiation of VSMCs via ERK signal pathway.