The experimental study of HMGB1 expression in macrophage induced by LPS

FENG Hua-guo; et al

doi:10.13406/j.cnki.cyxb.2010.07.045

您当前的位置：

首页 >

文章列表页 >

The experimental study of HMGB1 expression in macrophage induced by LPS

更新时间：2024-01-17

- The experimental study of HMGB1 expression in macrophage induced by LPS
- Journal of Chongqing Medical University Vol. 35, Issue 7, Pages: 999-1003(2010)
- 作者机构：
  
  1. 重庆医科大学附属第二医院肝胆外科
  2. 重庆医科大学病毒性肝炎研究所
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.07.045
  CLC： R459.7
- Published：2010，
扫描看全文
[1]冯华国,孙航,吴传新,郭晖,龚建平,刘杞,李生伟.内毒素诱导巨噬细胞HMGB1表达的实验研究[J].重庆医科大学学报,2010,35(07):999-1003.

FENG Hua-guo, et al. The experimental study of HMGB1 expression in macrophage induced by LPS. [J]. Journal of Chongqing Medical University 35(7):999-1003(2010)
[1]冯华国,孙航,吴传新,郭晖,龚建平,刘杞,李生伟.内毒素诱导巨噬细胞HMGB1表达的实验研究[J].重庆医科大学学报,2010,35(07):999-1003. DOI： 10.13406/j.cnki.cyxb.2010.07.045.

FENG Hua-guo, et al. The experimental study of HMGB1 expression in macrophage induced by LPS. [J]. Journal of Chongqing Medical University 35(7):999-1003(2010) DOI： 10.13406/j.cnki.cyxb.2010.07.045.

摘要

目的:探讨内毒素（脂多糖

Lipoplysaccharide

LPS）刺激巨噬细胞后高迁移率族蛋白B1（High mobility group protein B1

HMGB1）的释放和表达水平。方法:用100ng/ml的LPS刺激RAW264.7细胞

分别于刺激后0、6、12、18、24、30、36h和48h

用Western blot检测细胞培养上清、细胞浆和细胞核内HMGB1蛋白的含量;用RT-PCR检测细胞中HMGB1的mRNA表达情况;用免疫细胞化学、激光共聚焦显微镜观察LPS刺激后细胞浆和胞核内HMGB1的含量变化。分别于LPS刺激后0、1、2、4h和6h

用酶联免疫吸附实验（Enzymelinked immunoassay

ELISA）测定细胞培养上清中TNF-α、IL-6的含量。结果:LPS刺激后0～12h细胞培养上清中无法检测到HMGB1蛋白

18h后细胞培养上清中HMGB1蛋白的含量开始增加

于24h达到高峰

以后稳定维持在较高水平;LPS刺激前

HMGB1主要分布于细胞核内

胞核中HMGB1蛋白含量较高

胞浆中HMGB1蛋白含量少;LPS刺激后12～48h细胞浆中HMGB1蛋白含量逐渐增加;而LPS刺激后12～24h细胞核内HMGB1含量逐渐减少

30h后细胞核内HMGB1含量又逐渐增多。RT-PCR结果显示

LPS刺激后0～18h

培养细胞中HMGB1的mRNA表达量无明显变化

24～36h培养细胞中HMGB1的 mRNA表达量明显增加。ELISA结果显示

LPS刺激后1h

细胞培养上清中TNF-α、IL-6的含量明显增高

2h达到高峰。结论:LPS可诱导单核-巨噬细胞内HMGB1、TNF-α、IL-6表达和释放增加

LPS诱导巨噬细胞释放HMGB1的时间明显晚于TNF-α、IL-6的释放时间;LPS诱导巨噬细胞内的HMGB1从细胞核向细胞浆转位

并释放到细胞外;LPS诱导巨噬细胞HMGB1 mRNA基因表达上调的时间和蛋白合成的时间出现较晚。

Abstract

Objective:To elucidate the release and expression of high mobility group protein B1(HMGB1)in macrophage induced by Lipoplysaccharide(LPS).Methods:The murine macrophage-like cell line RAW264.7 was stimulated by LPS.The contents of HMGB1 protein were detected by Western blot in cultured cells supernatant

cytoplasm and nucleus.The expression of HMGB1 mRNA in cultured cell was determined by RT-PCR.The HMGB1 contents in cytoplasm and nucelus were observed using immunocytochemistry and laser confocal scanning microscopy(LCSM).Results:The contents of HMGB1 protein in supernatant could not be detected at 0～12 h

but increased gradually from 18h

reached a peak at 24 h and maintained stably at high level after the stimulation of LPS.The level of HMGB1 protein in cytoplasm increased gradually from 0 h to 48 h.The HMGB1 protein level in nucleus was higher at 0 h and decreased gradually at 12～24 h

but increased gradually at 36～48h after LPS stimulated.The expression of HMGB1 mRNA was not significantly changed at 0～18 h but enhanced significantly at 24～36 h after LPS stimulated.Conclusion:The release of HMGB1

TNF-αand IL-6 increased in macrophages induced by LPS.HMGB1 was released later than that of TNF-αand IL-6.The expression of HMGB1 mRNA is delayed. HMGB1 released to outside of the cells was come from the prior synthetic reservation in nucleus at early time.Lately new HMGB1 protein was synthesized in nucleus and was transferred into cytoplasm

which led to the level of HMGB1 in nucleus and in cytoplasm increased.

关键词

高迁移率族蛋白B1脓毒症内毒素

Keywords

High mobilitygroup protein B1sepsisLPS

references

Views

332

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Establishment of a multiplex PCR system for rapid detection of sepsis

Role of T lymphocytes apoptosis in immunosuppression and prognosis of sepsis

Effects of obesity on liver injury and inflammatory response in septic mouse model

Clinical study of immunoregulation therapy in the patients with severe trauma,sepsis or mods

The relationship between gene expression of Toll-like receptor 2 and acute kidney lesion in mice with sepsis

Related Author

Wei Li

Bo Pang

Pengpeng Wang

Jian Wu

Yan Zeng

Yang Tao

Yuanyuan Zhang

Xun Li

Related Institution

School of Medicine，Chongqing University

Department of Laboratory and Critical Care Medicine，The Affiliated Central Hospital of Chongqing University/Chongqing Emergency Medical Center

MyGenostics Inc

Department of Nephrology,The First Affiliated Hospital of Chongqing Medical University

Department of Emergency,The First Affiliated Hospital of Chongqing Medical University

⁰