| 何 飞,仇 伟,张雅鸥,袁 理,陈金数,张国权.Notch信号及自噬在三氧化矿化聚合物促人牙髓细胞体外增殖中的作用[J].,2017,17(24):4635-4638 |
| Notch信号及自噬在三氧化矿化聚合物促人牙髓细胞体外增殖中的作用 |
| Effect of Notch Signal and Autophage on MTA induced Proliferation of Human Dental Pulp Cells in Vitro |
| 投稿时间：2017-02-18 修订日期：2017-03-07 |
| DOI：10.13241/j.cnki.pmb.2017.24.008 |
| 中文关键词: 三氧化矿化聚合物 牙髓细胞 增殖 Notch 自噬 |
| 英文关键词: Mineral trioxide aggregate Dental pulp cells (DPCs) Proliferation Notch Autophagy |
| 基金项目:深圳市科创委科技计划项目（JCYJ201416122811975） |
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| 中文摘要: |
| 摘要 目的：探讨Notch信号及自噬在三氧化矿化聚合物 (MTA) 促人牙髓细胞 (hDPCs) 体外增殖中的作用。方法：取临床上完整拔除的健康第三磨牙，通过酶消化法获得原代培养hDPCs。采用CCK-8比色法检测不同浓度 (0.5、1.0、2.0、5.0、10.0 mg/mL) MTA对hDPCs增殖的影响，筛选出最佳促增殖作用浓度和作用时间。通过Western Blot法检测MTA作用下hDPCs Notch信号及自噬相关蛋白表达的变化。结果：1.0 mg/mL MTA对体外培养hDPCs促增殖作用最显著，高浓度MTA (10 mg/mL) 则具有一定的细胞毒性。与未处理hDPCs相比，MTA处理组hDPCs的Notch1、Hes1表达 (P＜0.05) 及自噬相关蛋白P62及LC3II/I表达 (P＜0.01) 均显著增高。平衡盐溶液 (EBSS) 饥饿实验诱导自噬发生后Notch1表达水平显著下降。结论：MTA可显著促进hDPCs的体外增殖，其作用机制可能与Notch1-Hes1信号转导途径激活及自噬抑制有关。 |
| 英文摘要: |
| ABSTRACT Objective: Mineral trioxide aggregate (MTA), a wildly used pulp capping material, could affect the proliferation and differentiation of dental pulp cells. The aim of this study is to study the roles of Notch signalling and autophagy in MTA induced human dental pulp cells (hDPCs) proliferation promotions. Methods: Healthy human third molars were collected and hDPCs were isolated by a combined digestion of collagenase I and dispase II. MTA extracts of different concentrations (0.5, 1.0, 2.0, 5.0, 10.0 mg/mL) were used to test the cytotoxicity by cells counting kit (CCK-8) assays and to select the optimum concentration for hDPCs survival. Expressions of Notch1, Hes1, LC3II / I and p62 in wild type and MTA treated hDPCs were detected by western blotting. Results: MTA extracts in a concentration of 1.0 mg/mL exerted most profoundly promotion effects on the proliferation of hDPCs among all concentrations tested. MTA of high concentration (10 mg/mL) was toxically to cells. Compared with that of wild type hDPCs, the expressions of Notch1 and Hes1(P＜0.05), or p62 and LC3II/I (P＜0.01) in MTA treated hDPCs were significantly increased. Much lower expression of Notch1 was detected in hDPCs when autophagy was induced by Earle's balanced salt solution (EBSS) starvation for 24 h. Conclusion: MTA could up-regulated hDPCs proliferation with highly relevant in stimulating Notch1-Hes1 signalling and inhibition of autophagy. The study is supposed to provide new insight in unrevealing the mechanisms of MTA mediated dental pulp cells proliferation. |
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