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

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8μg/mL衣霉素（TM）、0.5μmol毒胡萝卜素（TG）和0.8μmol 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、e...

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Published in上海交通大学学报（医学版） Vol. 32; no. 3; pp. 252 - 256
Main Author 刘卉芳 张惠洁 刘晓燕 胡其娴 马晓文 胡小磊 陈凤玲
Format Journal Article
LanguageChinese
Published 上海交通大学医学院附属第三人民医院内分泌科,上海,201900 2012
Subjects
内质网应激
巨噬细胞
类泛素折叠修饰蛋白
巨噬细胞
内质网应激
类泛素折叠修饰蛋白
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ISSN1674-8115
DOI10.3969/j.issn.1674-8115.2012.03.004

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Summary: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8μg/mL衣霉素（TM）、0.5μmol毒胡萝卜素（TG）和0.8μmol 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。
Bibliography:endoplasmic reticulum stress; macrophage; ubiquitin-fold modifier 1
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（P0.001）.After induction by TM or TG,the expression of Ufm1 mRNA and protein in RAW264.7 cells was significantly higher than
ISSN:1674-8115
DOI:10.3969/j.issn.1674-8115.2012.03.004