上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (3): 252-.doi: 10.3969/j.issn.1674-8115.2012.03.004

• 论著（基础研究） • 上一篇    下一篇

内质网应激对巨噬细胞类泛素折叠修饰蛋白Ufm1表达的影响

刘卉芳, 张惠洁, 刘晓燕, 胡其娴, 马晓文, 胡小磊, 陈凤玲   

  1. 上海交通大学 医学院附属第三人民医院内分泌科, 上海 201900
  • 出版日期:2012-03-28 发布日期:2012-03-28
  • 通讯作者: 陈凤玲, 电子信箱: CFL1993@126.com。
  • 作者简介:刘卉芳（1984—）, 女, 硕士生；电子信箱: lhf_404@163.com。
  • 基金资助:

    国家自然科学基金（30870954）；上海交通大学医学院基金（YZ1054）；上海交通大学医学院新百人计划基金（09XBR01）；上海交通大学医学院附属第三人民医院基金（syz2010-02）

Effect of endoplasmic reticulum stress on expression of ubiquitin-fold modifier 1 in macrophages

LIU Hui-fang, ZHANG Hui-jie, LIU Xiao-yan, HU Qi-xian, MA Xiao-wen, HU Xiao-lei, CHEN Feng-ling   

  1. Department of Endocrinology, the Third People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 201900, China
  • Online:2012-03-28 Published:2012-03-28
  • Supported by:

    National Natural Science Foundation of China, 30870954；Shanghai Jiaotong University School of Medicine Foundation, YZ1054, 09XBR01；Foundation of the Third People's Hospital, Shanghai Jiaotong University School of Medicine, syz2010-02

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摘要：

目的 探讨内质网应激（ERS）对巨噬细胞类泛素折叠修饰蛋白（Ufm1）表达的影响。方法 制备糖尿病模型小鼠（db/db鼠，糖尿病组，n=6）和同窝野生型(WT)C57小鼠（对照组，n=6）的原代腹腔巨噬细胞，采用Real-Time PCR技术检测ERS相关分子（GRP78、XBP1）及Ufm1 mRNA的表达。分别用8 μg/mL衣霉素(TM)、0.5 μmol毒胡萝卜素(TG)和0.8 μmol TG诱导小鼠-单核巨噬细胞系RAW264.7，采用Real-Time PCR技术检测GRP78、XBP1和Ufm1 mRNA的表达，采用Western blotting法检测ERS相关因子（p-eIf2α、eIf2α、CHOP）及Ufm1蛋白的表达。结果 糖尿病组小鼠腹腔巨噬细胞GRP78、XBP1和Ufm1 mRNA的表达量均显著高于对照组（均P<0.001）。经TM或TG诱导的RAW264.7细胞中Ufm1 mRNA和蛋白的表达量均明显高于阴性对照细胞（P<0.05）。结论 糖尿病小鼠腹腔巨噬细胞中ERS相关因子及Ufm1的表达同时升高。体外诱导巨噬细胞株ERS能导致Ufm1的表达升高；提示Ufm1可能部分通过ERS途径影响巨噬细胞功能而参与糖尿病动脉粥样硬化过程。

关键词: 内质网应激, 巨噬细胞, 类泛素折叠修饰蛋白

Abstract:

Objective To investigate the effect of endoplasmic reticulum stress (ERS) on expression of ubiquitin-fold modifier 1 (Ufm1) in macrophages. Methods Peritoneal macrophages were obtained from mice with diabetes (db/db mice, diabetes group, n=6) and wild type (WT) C57 mice （control group, n=6）, and the expression of ERS-related factors (GRP78 and XBP1) and Ufm1 mRNA was detected by Real-Time PCR. RAW264.7 cells were induced with 8 μg/mL tunicamycin (TM), 0.5 μmol thapsigargin (TG) and 0.8 μmol TG separately, the expression of GRP78, XBP1 and Ufm1 mRNA was determined by Real-Time PCR, and the expression of ERS-related factors (p-eIf2α, eIf2α and CHOP) and Ufm1 protein was detected by Western blotting. Results The expression of GRP78, XBP1 and Ufm1 mRNA in peritoneal macrophages of diabetes group was significantly higher than that of control group (P<0.001). After induction by TM or TG, the expression of Ufm1 mRNA and protein in RAW264.7 cells was significantly higher than that in negative controls (P<0.05). Conclusion The expression of both ERS-related factors and Ufm1 in peritoneal macrophages increases in mice with diabetes. ERS in macrophages induced in vitro may increase the expression of Ufm1, which indicates that Ufm1 may participate in diabetic atherosclerosis by influencing function of macrophages partially through ERS pathway.

Key words: endoplasmic reticulum stress, macrophage, ubiquitin-fold modifier 1