HSF1调控的炎症相关基因的筛选及SOCS3基因的实验验证  

Screen of inflammatory genes regulated by heat shock factor 1 and corroboration with SOCS3 gene

作　　者：于凤秀[1] 张华莉[1] 陈广文[1] 王秋鹏[1] 石永忠[1] 刘瑛[1] 梁秋娟[1] 肖献忠[1] 

机构地区：[1]中南大学湘雅医学院病理生理学教研室,长沙410078

出　　处：《中南大学学报（医学版）》2006年第2期167-173,共7页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(30330280);国家自然科学基金(30371382);湖南省自然科学基金(03jjY3050)

摘　　要：目的:筛选热休克因子1(HSF1)调控的炎症相关基因,初步探讨HSF1对下游基因的调控作用。方法:采用HSF1基因敲除小鼠,制备内毒素腹腔注射以及热休克反应后再给予内毒素腹腔注射的内毒素血症模型,分别抽提HSF1缺失突变纯合子和HSF1野生型小鼠肺组织总RNA,用微阵列进行筛选;用转录元件搜索软件分析差异表达基因的启动子区;选取启动子区含有热休克元件的基因:细胞因子信号转导抑制子3(SOCS3),采用热休克反应(HSR)和HSF1过表达的RAW 2 6 4.7巨噬细胞给予内毒素刺激,抽提总RNA进行RT-PCR,Northern印迹实验,观察HSR和HSF1过表达对内毒素诱导的SOCS3基因表达的影响。结果:用微阵列的方法筛选到HSF1可能抑制表达的炎症介质基因1 5个,其中9个基因的启动子区含有热休克元件;HSF1可能促进表达的炎症介质基因1 1个,其中8个基因的启动子区含有热休克元件;HSR和HSF1过表达可明显抑制小鼠RAW 2 6 4.7巨噬细胞内毒素诱导的SOCS3mRNA的表达。结论:HSF1可抑制炎症介质SOCS3mRNA的表达。Objective To screen the inflammatory mediators genes regulated by HSF1, and explore the mechanism of downstream genes regulated by HSF 1. Methods HSF 1 ^-/- and HSF 1^＋/＋ mice were injected with 15 mg/kg LPS intraperitoneally （ip） , respectively, and were treated as previous after HSR. The total RNA of lung tissues were extracted and fittrated by SuperArray gene Microarry. The promoter of candidate genes were analyzed by transcription element search software to search for heat shock element （ HSE ）. Select the suppressor of cytokine signaling 3 （ SOCS3 ） with HSE. Macrophage cells were stimulated with 400 ng/mL LPS, and were treated as previous after HSR, then the total RNA was extracted respectively. RT-PCR and northern blot assay were performed to detect the expression levels of SOCS3 mRNA. Results Fifteen genes were repressed by HSF 1 , including 9 genes with complete HSE. Eleven genes were accelerated by HSF1 possibly, including 8 genes with complete HSE. The promoter of SOCS3 gene contained one complete HSE. LPS stimulation obviously increased the levels of SOCS3 mRNA in macrophages of RAW264. 7 mice, which was inhibited by HSR and over-expression of HSF1. Conclusion HSR or HSF1 inhibits LPS induced expression of SOCS3 mRNA; HSF1 might inhibit LPS-induced expression of SOCS3 mRNA by binding to HSE in the promoter of SOCS3 gene.

关 键 词：热休克反应 热休克因子1 细胞因子信号转导抑制子3 

分 类 号：R363[医药卫生—病理学]