Institute of Stomatology, Nanjing Medical University, Nanjing, Jiangsu, 210029, China; Endodontic Department, Suzhou Stomatological Hospital, Suzhou, Jiangsu, 215005, China.
Cell Prolif. 2013 Dec;46(6):677-84. doi: 10.1111/cpr.12071. Epub 2013 Oct 24.
Oestrogen has been proven to significantly enhance osteogenic potency, while oestrogen deficiency usually leads to impaired osteogenic differentiation of mesenchymal stem cells. However, little is known concerning direct effects of oestrogen on differentiation of human dental pulp stem cells (DPSCs).
In this study, human DPSCs were isolated and treated with 10(-7) m 17β-oestradiol (E2). Alkaline phosphatase (ALP) assay and alizarin red staining were performed.
Alkaline phosphatase and alizarin red showed that E2 treatment significantly enhanced ALP activity and mineralization ability of DPSCs, but had no effect on cell proliferation. Real-time RT-PCR and western blot assay demonstrated that odonto/osteogenic markers (ALP, RUNX2/RUNX2, OSX/OSX, OCN/OCN and DSPP/DSP) were significantly upregulated in the cells after E2 treatment. Moreover, phosphorylation of cytoplasmic IκBα/P65 and expression of nuclear P65 were enhanced in a time-dependent manner following E2 treatment, suggesting activation of NF-κB signaling. Conversely, inhibition of the NF-κB pathway suppressed E2-mediated upregulation of odonto/osteogenic markers, indicating that the NF-κB pathway was pivotal for E2-mediated differentiation.
These findings provide evidence that 10(-7) m 17β-oestradiol promoted odonto/osteogenic differentiation of human DPSCs via activation of the NF-κB signaling pathway.
雌激素已被证明能显著增强成骨潜能,而雌激素缺乏通常会导致间充质干细胞成骨分化受损。然而,关于雌激素对人牙髓干细胞(DPSCs)分化的直接影响知之甚少。
在这项研究中,分离并培养人牙髓干细胞,用 10(-7) m 17β-雌二醇(E2)处理。进行碱性磷酸酶(ALP)检测和茜素红染色。
碱性磷酸酶和茜素红显示,E2 处理显著增强了 DPSCs 的 ALP 活性和矿化能力,但对细胞增殖没有影响。实时 RT-PCR 和 Western blot 分析表明,E2 处理后牙/成骨标志物(ALP、RUNX2/RUNX2、OSX/OSX、OCN/OCN 和 DSPP/DSP)在细胞中显著上调。此外,E2 处理后,细胞质 IκBα/P65 的磷酸化和核 P65 的表达呈时间依赖性增强,提示 NF-κB 信号通路被激活。相反,NF-κB 通路的抑制抑制了 E2 介导的牙/成骨标志物的上调,表明 NF-κB 通路对于 E2 介导的分化至关重要。
这些发现为 10(-7) m 17β-雌二醇通过激活 NF-κB 信号通路促进人牙髓干细胞牙/成骨分化提供了证据。