摘要
目的研究肿瘤坏死因子α(TNFα)对小鼠纤维母细胞(L929)中葡萄糖调节蛋白78(glucose regulatedprotein78,GRP78)、X盒结合蛋白1(X box binding protein 1,XBP-1)表达的影响,探讨TNFα信号传导通路与内质网应激的相关性。方法体外培养L929细胞,分别用TNFα、Tunicamycin作用0、24、、68、和10 h,以Western Blot、Northern Blot检测内质网应激的标志分子:GRP78蛋白的表达,以及非折叠蛋白反应中的关键分子:XBP-1蛋白及mRNA水平表达;用RT-PCR结合Pst1酶切法检测XBP-1 mRNA的剪接作用。结果TNFα作用于L929细胞即可以引起GRP78蛋白、XBP-1蛋白及XBP-1mRNA表达的升高,又可以诱导XBP-1mRNA的剪接作用;且这些分子表达的升高及mRNA的剪接作用均发生在TNFα作用的早期(开始于TNFα作用的2 h,4 h达到高峰,6 h后逐渐减弱)。结论TNFα作用于L929细胞可以诱导内质网应激,由此引起的活化转录因子6(activating transcription factor 6,ATF6)、ER转膜蛋白激酶1(inositol-requiring enzyme 1,IRE1)介导的未折叠蛋白反应对细胞具有促生存作用。
Objective To investigate the association of TNFα signal pathway with endoplasmic reticulum stress （ER Stress）, the expression of glucose regulated protein78 （GRP78）and X box binding protein 1 （XBP-1） were examined in tumor necrosis factor α （TNFα）-treated L929 cells. Methods L929 cells were stimulated with TNFα and Tunicamycin for 0 - 10 h. The expression of ER chaperone GRP78 protein,XBP-I protein and XBP-1 mRNA were detected by Western blotting and Northern blotting. The splicing of XBP-1 mRNA were analyzed by RT-PCR with Pstl digestion. Results TNFα up-regulated the expression of GRP78 protein and increased the expression of XBP-1 in protein and mRNA levels. TNFα further induced the splicing of XBP-1 mRNA. All these changes happened in the early phage of TNFα stimulation （ it began at 2 h, peaked at 4 h and decreased gradually after 6 h） . Conclusion TNFzt induced the ER stress in L929 cells and subsequently initiated the unfolded protein response mediated by ATF6 and IRE1, which contributes to protection of cell from TNFα stimulation.
出处
《毒理学杂志》
CAS
CSCD
北大核心
2008年第6期431-434,共4页
Journal of Toxicology
基金
人事部留学人员科技活动项目择优资助经费(2007LHR01)
中国中医科学院基本科研业务费自主选题项目(ZZ2006015)
作者简介
薛欣，博士，研究方向：免疫生物学。
通讯作者：赵京元，博士，研究方向：免疫病理学。
