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

作　　者：何飞[1] 仇伟[2] 张雅鸥[2] 袁理[1] 陈金数 张国权[1] 

机构地区：[1]深圳市人民医院口腔科暨南大学第二附属医学院,广东深圳518020 [2]健康科学与技术重点实验室生命与健康学部清华大学深圳研究生院,广东深圳518055

出　　处：《现代生物医学进展》2017年第24期4635-4638,4718,共5页Progress in Modern Biomedicine

基　　金：深圳市科创委科技计划项目(JCYJ201416122811975)

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(h DPCs)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h DPCs。采用CCK-8比色法检测不同浓度(0.5、1.0、2.0、5.0、10.0 mg/m L)MTA对h DPCs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h DPCs Notch信号及自噬相关蛋白表达的变化。结果:1.0 mg/m L MTA对体外培养h DPCs促增殖作用最显著,高浓度MTA(10 mg/m L)则具有一定的细胞毒性。与未处理h DPCs相比,MTA处理组h DPCs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h DPCs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。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.Conclusions：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.

关 键 词：三氧化矿化聚合物 牙髓细胞 增殖 NOTCH 自噬 

分 类 号：R-33[医药卫生] R780.2