Notch信号及自噬在三氧化矿化聚合物促人牙髓细胞体外分化中的作用  

Effect of Notch signaling and autophagy on MTA induced differentiation of human dental pulp cells in vitro

作　　者：何飞[1] 郑雨燕[1] 仇伟[2] 张国权[1] 麦穗[3] HE Fei;ZHENG Yuyan;QIU Wei;ZHANG Guoquan;MAI Sui(Department of Stomatology, the Second Clinical Medical College & Shenzhen People..s Hospital, Ji..nan University, Shenzhen 518020, China;Key Lab in Healthy Science and Technology, Division of Life Science, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China;Department of Conservative Dentistry and Endodontics Guanghua School of Stomatology, Sun Yat.sen University, Guangzhou 510055, China)

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]清华大学深圳研究生院生命与健康学部健康科学与技术重点实验室,广东深圳518055 [3]中山大学光华口腔医学院.附属口腔医院牙体牙髓科,广东广州510055

出　　处：《口腔疾病防治》2017年第7期414-419,共6页Journal of Prevention and Treatment for Stomatological Diseases

基　　金：广东省自然科学基金项目(2014A030313068);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(mineral trioxide aggregate,MTA)促人牙髓细胞(human dental pulp cells,h DPCs)体外分化的作用。方法体外培养h DPCs,采用荧光定量PCR法检测MTA作用不同时间(24 h、3 d、7 d)对h DPCs碱性磷酸酶(alkaline phosphatase,ALP)、核心结合蛋白因子2(runt-related transcription factor 2,Runx2)及牙本质涎磷蛋白(dentin sialophoprotein,DSPP)表达的影响;Von Kossa染色观察细胞钙化结节形成的变化;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0.1 mg/m L MTA即可促进体外培养h DPCs的分化。与未处理h DPCs的空白对照组相比,MTA组h DPCs的Notch信号成分Notch1、Hes1、Jagged1及自噬相关蛋白p62表达均明显增高,差异均有统计学意义(P<0.05)。以自噬体与溶酶体融合抑制剂Bafilomycin A1(BAF)刺激细胞,自噬潮受抑制,Notch1表达升高,MTA具有类似作用。结论 MTA可显著促进h DPCs的体外分化,其作用机制可能与Notch1-Jagged1-Hes1信号转导途径激活及自噬抑制有关。Objective The aim of this study is to investigate the roles of Notch signaling and autophagy on mineraltrioxide aggregate(MTA)induced differentiation of human dental pulp cells(hDPCs).Methods Third molars fromhealthy human were collected and hDPCs were isolated by a combined digestion of collagenaseⅠand dispaseⅡ.Realtime PCR were used to test the mRNA expression levels of alkaline phosphatase(ALP),runt?related transcription factor2(Runx2)and dentin sialophoprotein(DSPP)in MTA treated hDPCs in different time(24h,3d and7d).The mineral?ization nodules formed by hDPCs with or without MTA treatment were detected by Von Kossa staining.Expressions ofNotch1,Jagged1,Hes1,LC3Ⅱ/LC3Ⅰand p62in wild type and MTA treated hDPCs were detected by western blotting.Results MTA extracted in a concentration of0.1mg/mL could promote the differentiation of hDPCs.Compared withthat of wild type hDPCs,the expressions of Notch1,Hes1,or Jagged1and p62(P<0.01)in MTA treated hDPCs weresignificantly increased.MTA treatment showed inhibition effects on autophagy flux similar to Bafilomycin A1,a specificinhibitor of fusion between autophagosomes and lysosomes.Conclusion MTA could promote hDPCs differentiationwith highly relevant in stimulating Notch1?Jagged1?Hes1signaling and inhibition of autophagy flux.

关 键 词：三氧化矿化聚合物 牙髓细胞 分化 NOTCH 自噬 

分 类 号：R780.2[医药卫生—口腔医学]