Simvastatin promotes osteogenic differentiation of mouse embryonic stem cells via canonical Wnt/β-catenin signaling
Simvastatin promotes osteogenic differentiation of mouse embryonic stem cells via canonical Wnt/β-catenin signaling
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DOI：
10.1007/s10059-011-0107-6
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发表时间：
2011-11-01
影响因子：
3.8
通讯作者：
Heo, Jung Sun
中科院分区：
文献类型：
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作者：
Qiao, Ling Juan;Kang, Kyung Lhi;Heo, Jung Sun
辛伐他汀是3-羟基-3-甲基戊二酰辅酶A（HMG-CoA）还原酶的抑制剂，已知可减少胆固醇的生物合成。然而，最近的研究表明，辛伐他汀显示出多种胆固醇非依赖性功能，包括细胞分化。在这项研究中，我们研究了辛伐他汀对小鼠胚胎干细胞（ESCs）成骨分化的刺激作用。辛伐他汀的成骨作用在相对较低的剂量（范围从1 nM到200 nM）下观察到。在添加辛伐他汀的成骨培养基中孵育ESC在第7天显著增加碱性磷酸酶（ALP）活性。辛伐他汀孵育14天后，基质矿化也增加，并显示出关键水平。通过上调成骨转录因子Runx 2、OSX和OCN的mRNA表达来确定辛伐他汀诱导的ESCs成骨分化。此外，OCN、骨桥蛋白（OPN）和I型胶原（科尔I）的蛋白表达增加采用蛋白质印迹分析和免疫细胞化学进行评估。然而，通过DKK-1阻断经典Wnt信号转导下调辛伐他汀诱导的ALP活性和每个成骨转录因子的mRNA表达。此外，β-catenin特异性siRNA转染降低了OCN、OPN和科尔I的蛋白水平。总的来说，这些发现表明，辛伐他汀通过激活经典Wnt/β-连环蛋白信号传导增强了ESC向成骨谱系的分化。
Simvastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, has been known to reduce cholesterol biosynthesis. However, recent studies demonstrate that simvastatin shows diverse cholesterol-independent functions including cellular differentiation. In this study, we investigated the stimulatory effect of simvastatin on the osteogenic differentiation of mouse embryonic stem cells (ESCs). The osteogenic effect of simvastatin was observed at relatively low doses (ranging from 1 nM to 200 nM). Incubation of ESCs in simvastatin-supplemented osteogenic medium significantly increased alkaline phosphatase (ALP) activity at day 7. The matrix mineralization was also augmented and demonstrated pivotal levels after 14 days incubation of simvastatin. Osteogenic differentiation of ESCs by simvastatin was determined by upregulation of the mRNA expression of runtrelated gene 2 (Runx2), osterix (OSX), and osteocalcin (OCN) as osteogenic transcription factors. Moreover, the increased protein expression of OCN, osteopontin (OPN), and collagen type I (Coll I) was assessed using Western blot analysis and immunocytochemistry. However, the blockage of canonical Wnt signaling by DKK-1 downregulated simvastatin-induced ALP activity and the mRNA expression of each osteogenic transcription factor. Furthermore, the beta-catenin specific siRNA transfection decreased the protein levels of OCN, OPN, and Coll I. Collectively, these findings suggest that simvastatin enhances the differentiation of ESCs toward osteogenic lineage through activation of canonical Wnt/beta-catenin signaling.