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

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,s XBP1)、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)m RNA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。

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 m RNA 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.