The role of sirtuin 1 in osteoblastic differentiation in human periodontal ligament cells
The role of sirtuin 1 in osteoblastic differentiation in human periodontal ligament cells
复制标题
DOI：
10.1111/j.1600-0765.2011.01394.x
复制
发表时间：
2011-12-01
影响因子：
3.5
通讯作者：
Kim, E. -C.
中科院分区：
文献类型：
--
作者：
Lee, Y. -M.;Shin, S. -I.;Kim, E. -C.
背景和目的：sirtuin 1（SIRT 1）的激活促进角质形成细胞和间充质干细胞的分化，但抑制肌肉和脂肪细胞的分化。然而，SIRT 1参与人牙周膜细胞向成骨细胞样细胞的分化仍不清楚。为了确定SIRT 1在人牙周膜细胞中的作用，我们检测了人牙周膜细胞成骨分化过程中SIRT 1 mRNA和SIRT 1蛋白的水平。此外，我们还研究了SIRT 1过表达和低表达对人牙周膜细胞分化的影响，以及所涉及的信号机制。材料和方法：通过RT-PCR、real-time PCR、茜素红染色和western blotting检测SIRT 1和成骨细胞分化标志物的表达。在成骨诱导培养基（OM）中生长3 d的细胞中观察到SIRT 1 mRNA和SIRT 1蛋白的显著上调。白藜芦醇和异烟酰胺激活SIRT 1以剂量依赖性方式刺激成骨细胞分化，通过表达编码碱性磷酸酶、骨桥蛋白、骨钙素、osterix和Runx 2的mRNA进行评估，并诱导钙沉积。相反，使用sirtinol，烟酰胺和RNA干扰的基因沉默抑制SIRT 1抑制矿化和成骨细胞标志物mRNA的表达。进一步的机制研究表明，白藜芦醇处理增加Akt，腺苷一磷酸激酶（AMPK），Smad 1/5/8和c-Jun N-末端激酶的磷酸化，但减少OM诱导的核因子-κ B的激活。相反，sirtinol的应用抑制Akt，AMPK，Smad 1/5/8，p38，ERK和c-Jun N-末端激酶的磷酸化，并增强核因子jB活性，在OM-刺激cells.Conclusion：这些数据表明，SIRT 1是一个有效的调节人牙周膜细胞的分化，并可能具有临床意义的牙周骨再生。
Background and Objective: Activation of sirtuin 1 (SIRT1) promotes the differentiation of keratinocytes and mesenchymal stem cells, but inhibits the differentiation of muscle and fat cells. However, the involvement of SIRT1 in the differentiation of human periodontal ligament cells into osteoblast-like cells remains unclear. To identify the role of SIRT1 in human periodontal ligament cells, we measured SIRT1 mRNA and SIRT1 protein levels during the osteoblastic differentiation of human periodontal ligament cells. Additionally, we investigated the effects of overexpressing and underexpressing SIRT1 on the differentiation of human periodontal ligament cells, and the signaling mechanisms involved.Material and Methods: Expression of SIRT1 and osteoblastic differentiation markers was assessed by RT-PCR, real-time PCR, Alizarin red staining and western blotting.Results: Marked upregulation of SIRT1 mRNA and SIRT1 protein was observed in cells grown for 3 d in osteogenic induction medium (OM). Activation of SIRT1 using resveratrol and isonicotinamide stimulated osteoblastic differentiation in a dose-dependent manner, as assessed by the expression of mRNAs encoding alkaline phosphatase, osteopontin, osteocalcin, osterix and Runx2, and induced calcium deposition. In contrast, inhibition of SIRT1 using sirtinol, nicotinamide and gene silencing by RNA interference suppressed mineralization and the expression of osteoblast marker mRNAs. Further mechanistic studies revealed that resveratrol treatment increased the phosphorylation of Akt, adenosine monophosphate kinase (AMPK), Smad 1/5/8 and c-Jun N-terminal kinase, but reduced OM-induced activation of nuclear factor-kappa B. Conversely, application of sirtinol suppressed the phosphorylation of Akt, AMPK, Smad 1/5/8, p38, ERK and c-Jun N-terminal kinase, and enhanced nuclear factor-jB activity, in OM-stimulated cells.Conclusion: These data suggest that SIRT1 is a potent regulator of differentiation of human periodontal ligament cells and may have clinical implications for periodontal bone regeneration.