衣霉素诱导牙髓细胞内质网应激模型的建立

doi:10.19439/j.sjos.2018.02.005

上海口腔医学 ›› 2018, Vol. 27 ›› Issue (2): 135-138.doi: 10.19439/j.sjos.2018.02.005

衣霉素诱导牙髓细胞内质网应激模型的建立

李莉芬, 文扬, 江龙^*, 朱亚琴^*

上海交通大学医学院附属第九人民医院·口腔医学院口腔综合科,国家口腔医学疾病临床研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海 200011

收稿日期:2017-09-08 修回日期:2017-10-24 出版日期:2018-04-25 发布日期:2018-05-14
通讯作者: 朱亚琴,E-mail: zyq1590@163.com;江龙,E-mail:jianglong25a@163.com。*共同通信作者
作者简介:李莉芬（1986-）,女,博士,住院医师, E-mail:yuyo2013@126.com
基金资助:
国家自然科学基金（81271134,81300867）; 上海高校高峰高原学科建设项目; 上海交通大学医学院附属第九人民医院院级基金（2016-11）

Establishment of a model of endoplasmic reticulum stress response in dental pulp cells induced by tunicamycin

LI Li-fen, WEN Yang, JIANG Long, ZHU Ya-qin

Department of General Dentistry, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; National Clinical Research Center of Stomatology; Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology. Shanghai 200011, China

Received:2017-09-08 Revised:2017-10-24 Online:2018-04-25 Published:2018-05-14

摘要/Abstract

摘要： 目的:用衣霉素诱导人牙髓细胞（dental pulp cells, DPCs）,构建人DPCs内质网应激模型,为进一步探讨内质网应激介导人DPCs相关疾病的分子机制提供实验基础。方法:改良组织块法体外培养人DPCs;MTT法检测不同浓度衣霉素对人DPCs活性的影响;PCR检测人DPCs内质网应激关键分子的表达。采用SPSS17.0软件包对数据进行统计学分析。结果:随着衣霉素浓度的增加,人DPCs存活率逐渐下降。衣霉素诱导后,人DPCs内质网应激关键分子剪切x盒结合蛋白1(splicing x-box binding protein-1, sXBP1)、活化转录因子4（activating transcription factor 4,ATF4）、葡萄糖调节蛋白质78（glucose-regulated protein 78, GRP78）以及C/EBP同源蛋白(C/EBP homologous protein, CHOP) mRNA表达水平升高。结论:衣霉素诱导人DPCs内质网应激发生,模型成功建立。

关键词: 衣霉素, 牙髓细胞, 内质网应激

Abstract: PURPOSE: The aim of this study was to establish a model of endoplasmic reticulum (ER) stress in dental pulp cells(DPCs) induced by tunicamycin to better understand the molecular mechanism of DPCs related diseases mediated by ER stress. METHODS: DPCs were cultured using modified tissue explant technique in vitro and cultured in presence or absence of tunicamycin. DPCs' viability was measured by methylthiazol tetrazolium (MTT) assay. The mRNA level of ER stress markers was examined by RT-PCR. The data were analyzed with SPSS17.0 software package. RESULTS: The proliferative ability of DPCs decreased when exposed to tunicamycin in a dose-dependent manner. Treatment with tunicamycin resulted in up-regulation of ER stress genes, such as splicing x-box binding protein-1(sXBP1), activating transcription factor 4(ATF4), glucose-regulated protein 78(GRP78) and C/EBP homologous protein (CHOP). CONCLUSIONS: The results indicate that ER stress response is induced in DPCs by tunicamycin, and the ER stress model is successfully established.

Key words: Tunicamycin, Dental pulp cells, ER stress

中图分类号:

R781.4

李莉芬, 文扬, 江龙, 朱亚琴. 衣霉素诱导牙髓细胞内质网应激模型的建立[J]. 上海口腔医学, 2018, 27(2): 135-138.

LI Li-fen, WEN Yang, JIANG Long, ZHU Ya-qin. Establishment of a model of endoplasmic reticulum stress response in dental pulp cells induced by tunicamycin[J]. Shanghai Journal of Stomatology, 2018, 27(2): 135-138.

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衣霉素诱导牙髓细胞内质网应激模型的建立

Establishment of a model of endoplasmic reticulum stress response in dental pulp cells induced by tunicamycin

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