Estrogen receptor mediates simvastatin-stimulated osteogenic effects in bone marrow mesenchymal stem cells
Estrogen receptor mediates simvastatin-stimulated osteogenic effects in bone marrow mesenchymal stem cells
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DOI：
10.1016/j.bcp.2015.09.018
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发表时间：
2015-12-01
影响因子：
5.8
通讯作者：
Chang, Je-Ken
中科院分区：
文献类型：
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作者：
Chuang, Shu-Chun;Chen, Chung-Hwan;Chang, Je-Ken
辛伐他汀是一种HMG-CoA还原酶抑制剂，已知可促进成骨分化。然而，辛伐他汀诱导成骨的机制还不清楚。在这项研究中，我们假设雌激素受体（ER）介导辛伐他汀诱导的成骨分化。使用ER拮抗剂和siRNA来确定ER在辛伐他汀诱导的小鼠骨髓间充质干细胞（D1细胞）成骨中的参与。通过成骨标志物的mRNA表达、蛋白水平/活性和矿化来评价成骨。检测雌激素反应元件（ERE）启动子活性和ER-辛伐他汀结合亲和力。我们的研究结果表明，辛伐他汀诱导的成骨作用减少了治疗与ER α拮抗剂和ER α siRNA，但不通过G蛋白偶联雌激素受体（GPER-1）的拮抗剂特异性。辛伐他汀诱导的成骨作用通过E2处理进一步增加，并且通过ER拮抗剂或siRNA处理逆转。荧光素酶报告基因分析表明，辛伐他汀增加ER α依赖的转录活性，抑制ER α拮抗剂。此外，ER α-辛伐他汀结合试验显示，辛伐他汀的IC 50值为7.85 μ M，E-2的IC 50值为32.8 nM，表明辛伐他汀是ER α的弱配体。这些结果表明，辛伐他汀刺激的骨生成是由ER α介导的，而不是GPER-1。此外，这是第一份报告，以证明辛伐他汀作为ER配体和共激活剂，以增强ER α依赖的转录活性，从而促进骨生成。这些结果表明辛伐他汀诱导的骨生成是通过ER α依赖性途径介导的。(C)2015 Elsevier Inc. All rights reserved.
Simvastatin, an HMG-CoA reductase inhibitor, is known to promote osteogenic differentiation. However, the mechanism underlying simvastatin-induced osteogenesis is not well understood. In this study, we hypothesize that the estrogen receptor (ER) mediates simvastatin-induced osteogenic differentiation. ER antagonists and siRNA were used to determine the involvement of the ER in simvastatin-induced osteogenesis in mouse bone marrow mesenchymal stem cells (D1 cells). Osteogenesis was evaluated by mRNA expression, protein level/activity of osteogenic markers, and mineralization. The estrogen response element (ERE) promoter activity and the ER-simvastatin binding affinity were examined. Our results showed that the simvastatin-induced osteogenic effects were decreased by treatment with ER alpha antagonists and ER alpha siRNA but not by an antagonist specific for the G protein-coupled estrogen receptor (GPER-1). The simvastatin-induced osteogenic effects were further increased by E2 treatment and were reversed by ER antagonists or siRNA treatment. Luciferase reporter gene assays demonstrated that simvastatin increase ER alpha-dependent transcriptional activity that was suppressed by ER alpha antagonists. Furthermore, the ER alpha-simvastatin binding assay showed that IC50 value of simvastatin is 7.85 mu M and that of E-2 is 32.8 nM, indicating that simvastatin is a weak ligand for ER alpha. These results suggest that simvastatin-stimulated osteogenesis is mediated by ER alpha but not GPER-1. Moreover, this is the first report to demonstrate that simvastatin acts as an ER ligand and a co-activator to enhance ER alpha-dependent transcriptional activity and thus promotes osteogenesis. These results indicate that simvastatin-induced osteogenesis is mediated via an ER alpha-dependent pathway. (C) 2015 Elsevier Inc. All rights reserved.