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.

参考文献

[1] Ron D, Walter P.Signal integration in the endoplamic reticulum unfolded protein response[J]. Nat Rev Mol Cell Biol, 2007, 8(7): 519-529.
[2] Wei W, Gao Y, Wang C, et al.Excessive fluoride induces endoplasmic reticulum stress and interferes enamel proteinases secretion[J]. Environ Toxicol, 2013, 28(6): 332-341.
[3] Sharma R, Tsuchiya M, Bartlett JD.Fluoride induces endoplasmic reticulum stress and inhibits protein synthesis and secretion[J]. Environ Health Perspect, 2008, 116(9): 1142-1146.
[4] Kubota K, Lee DH, Tsuchiya M, et al.Fluoride induces endoplasmic reticulum stress in ameloblasts responsible for dental enamel formation[J]. J Biol Chem, 2005, 280(24): 23194-23202.
[5] Kim JW, Choi H, Jeong BC, et al.Transcriptional factor ATF6 is involved in odontoblastic differentiation[J]. J Dent Res, 2014, 93(5): 483-489.
[6] Diamanti E, Mathieu S, Jeanneau C, et al.Endoplasmic reticulum stress and mineralization inhibition mechanism by the resinous monomer HEMA[J]. Int Endod J, 2013, 46(2): 160-168.
[7] Oslowski CM, Urano F.Measuring ER stress and the unfolded protein response using mammalian tissue culture system[J]. Methods Enzymol, 2011, 490: 71-92.
[8] 江龙, 朱亚琴. 改良组织块法体外培养人牙周膜细胞和牙髓细胞[J]. 牙体牙髓牙周病学杂志, 2008, 18(4): 195-199.
[9] Jang WG, Kim EJ, Koh JT.Tunicamycin negatively regulates BMP2-induced osteoblast differentiation through CREBH expression in MC3T3E1 cells[J]. BMB Rep, 2011, 44(11): 735-740.
[10] 吴浩, 孟志超, 曹永平, 等. 内质网应激对膝关节炎大鼠关节软骨细胞的影响[J]. 中国组织工程研究, 2017, 21(16):2502-2508.
[11] Badiola N, Penas C, Minano-Molina A, et al.Induction of ER stress in response to oxygen-glucose deprivation of cortical cultures involves the activation of the PERK and IRE-1 pathway and of caspase-12[J]. Cell Death Dis, 2011, 2(4): e149.
[12] Oyadomari S, Araki E, Mori M.Endoplasmic reticulum stress mediated apoptosis in pancreatic beta-cells[J]. Apoptosis, 2002, 7(4): 335-345.
[13] Watson LM, Chan AK, Berry LR, et al.Overexpression of the 78-kDa glucose-regulated protein/immunoglobulin-binding protein (GRP78/Bip) inhibits tissue factor procoagulant activity[J]. J Biol Chem, 2003, 278(19): 17438-17447.
[14] Harding HP, Zhang Y, Bertolotti A, et al.Perk is essential for translational regulation and cell survival during the unfolded protein response[J]. Mol Cell, 2000, 5(5): 897-904.
[15] Guo HL, Hassa HM, Ding PP, et al.Pyrazinamide-induced hepatotoxicity is alleviated b 4-PBA via inhibition of the PERK-eIF2-ATF4-CHOP pathway[J]. Toxicology, 2017, 378: 65-75.

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

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	张安妮, 丁灿灿, 黄丽苹, 李适廷. 抑制连接蛋白43介导半通道活性促进脂多糖诱导的人牙髓细胞成牙本质分化[J]. 上海口腔医学, 2024, 33(1): 22-29.
[2]	王雪纯, 吴艳棋, 梅鹏, 张博钧, 朱敏. 低强度牵张应力对异丙肾上腺素诱导下牙周膜成纤维细胞炎症反应的影响[J]. 上海口腔医学, 2022, 31(3): 225-231.
[3]	徐方芳, 蒋备战. DKK1对脂多糖感染人牙髓细胞生物学行为的影响[J]. 上海口腔医学, 2021, 30(4): 355-359.
[4]	曹颖, 彭伟伟, 王璇, 伍甜甜, 杜嵘, 朱亚琴. 重组人结缔组织生长因子对牙髓细胞增殖与成牙本质分化的影响[J]. 上海口腔医学, 2020, 29(1): 7-12.
[5]	嵇国平, 田一宏, 刘美希, 沈莹, 沈刚. 内质网应激关键因子PREK在应力加载下成肌细胞凋亡中的作用及机制[J]. 上海口腔医学, 2019, 28(3): 259-263.
[6]	彭伟伟, 戴兆威, 曹颖, 韩俊力, 朱亚琴. 血清饥饿及融合培养对人牙髓细胞周期同步化和矿化活性的影响[J]. 上海口腔医学, 2019, 28(1): 1-5.
[7]	林田, 赵文青, 陆彦玲, 刘玉莹, 包丽荣, 吴煜. 人脐带间充质干细胞与牙髓细胞体外共培养对细胞生物学的影响[J]. 上海口腔医学, 2018, 27(4): 365-369.
[8]	李莉芬, 文扬, 江龙, 朱亚琴. 内质网应激在牙周膜细胞成骨分化中的表达[J]. 上海口腔医学, 2017, 26(6): 577-581.
[9]	包丽荣, 赵文青, 林田, 陆彦玲, 吴煜. BMP-2诱导人牙髓细胞分化过程中miRNA表达谱的变化[J]. 上海口腔医学, 2017, 26(5): 476-483.
[10]	何璇,韦维,陈文霞. 富血小板纤维蛋白凝胶三维结构及其对人牙髓细胞体外增殖的影响[J]. 上海口腔医学, 2015, 24(3): 263-268.
[11]	李健,张颖,马林,张凯强,雷双,顾何锋. 氟对大鼠成釉细胞GRP-78和caspase-12表达的影响[J]. 上海口腔医学, 2015, 24(1): 1-5.
[12]	王艳丽, 潘克清, 孙艳, 邓婧. 脂多糖对大鼠牙髓细胞ALP、BSP、DSPP表达的影响[J]. 上海口腔医学, 2014, 23(4): 431-435.
[13]	张凯强, 张颖, 刘璐, 顾何锋, 马林. 氟对大鼠切牙成釉细胞内质网伴侣分子表达的影响[J]. 上海口腔医学, 2013, 22(5): 481-486.
[14]	伍甜甜;杜嵘;李莉芬;朱亚琴. 应用流式细胞术检测人牙髓细胞内活性氧的方法探讨 [J]. 上海口腔医学, 2012, 21(6): 617-620.
[15]	魏福兰;耿杰;王春玲;王惠;张本君;张凡 . 牵张力作用下人牙髓细胞HIF-1α和VEGF的表达 [J]. 上海口腔医学, 2012, 21(5): 501-505.