School of Dentistry, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan.
Research Center of Tooth Bank and Dental Stem Cell Technology, Taipei Medical University, Taipei, Taiwan.
Int Endod J. 2018 Oct;51(10):1159-1170. doi: 10.1111/iej.12935. Epub 2018 Apr 26.
To evaluate the effect of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside (THSG) on cell proliferation and examine the mechanisms of THSG-enhanced proliferative potential in human dental pulp stem cells (hDPSC).
After treatment with THSG, hDPSC were collected. Cell viability was determined by MTS assay, while messenger RNA (mRNA) expressions of proliferation and stem cell markers were analyzed using real-time PCR. Flow cytometry was also conducted to analysis protein expression of stem cell markers. A colony-forming unit assay of hDPSC was carried out. Cellular telomerase activity was also identified using real-time PCR. In addition, proliferation-related proteins involved in the effects of THSG on hDPSC were analyzed by Western blotting. Data were analyzed using one-way analysis of variance and two-tailed Student's t-test.
Cell viability, colony-forming rates and telomerase activities of hDPSCs were enhanced after THSG treatment. mRNA expressions of proliferation markers (including expressions of NAD+-dependent histone deacetylase sirtuin 1 (SIRT1), proliferating cell nuclear antigen (PCNA), cyclin D1 and ribonucleotide reductase subunit M2 (RRM2)) increased significantly after THSG treatment (P < 0.05). Treatment with THSG for 3 h significantly augmented SIRT1 protein expression (P < 0.05). Furthermore, activities of proliferation-related proteins (including AMP-activated protein kinase (AMPK) and extracellular signal-regulated kinase (ERK) had also significantly increased at 3 h (P < 0.05). After THSG treatment, increased gene and protein expressions of pluripotent-like stem cell markers (including NANOG, OCT4, and SOX2) were observed.
2,3,5,4'-Tetrahydroxystilbene-2-O-β-glucoside treatment enhanced the renewal ability and proliferative potential of hDPSCs via the AMPK/ERK/SIRT1 axis, which may provide a novel autogenic cell-based therapeutic strategy in regenerative dentistry.
评估 2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷（THSG）对细胞增殖的影响，并研究 THSG 增强人牙髓干细胞（hDPSC）增殖潜能的机制。
用 THSG 处理 hDPSC 后收集细胞。通过 MTS 分析检测细胞活力，实时 PCR 分析增殖和干细胞标志物的信使 RNA（mRNA）表达。还通过流式细胞术分析干细胞标志物的蛋白表达。进行 hDPSC 集落形成单位分析。使用实时 PCR 还鉴定了细胞端粒酶活性。此外，通过 Western blot 分析了与 THSG 对 hDPSC 作用相关的增殖相关蛋白。数据采用单因素方差分析和双尾 Student's t 检验进行分析。
THSG 处理后，hDPSC 的细胞活力、集落形成率和端粒酶活性均增强。THSG 处理后，增殖标志物（包括 NAD+-依赖性组蛋白去乙酰化酶 SIRT1、增殖细胞核抗原（PCNA）、细胞周期蛋白 D1 和核糖核苷酸还原酶亚单位 M2（RRM2））的 mRNA 表达显著增加（P < 0.05）。THSG 处理 3 h 后 SIRT1 蛋白表达显著增加（P < 0.05）。此外，增殖相关蛋白（包括 AMP 激活的蛋白激酶（AMPK）和细胞外信号调节激酶（ERK））的活性在 3 h 时也显著增加（P < 0.05）。THSG 处理后，观察到多能样干细胞标志物（包括 NANOG、OCT4 和 SOX2）的基因和蛋白表达增加。
2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷处理通过 AMPK/ERK/SIRT1 轴增强了 hDPSC 的更新能力和增殖潜能，这可能为再生牙科提供一种新的自体细胞治疗策略。
