热休克反应对内毒素所致小鼠巨噬细胞炎症因子表达的抑制作用与MAPK信号转导通路的关系研究  

The Inhibition of Heat Shock Response on the Expression of Cytokines Induced by Lipopolysaccharide(LPS) in Murine Macrophages and Its Relation to MAPK Pathways

作　　者：石永忠[1] 唐道林[1] 王慷慨[1] 于凤秀[1] 刘梅冬[1] 张华莉[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,湖南长沙410078

出　　处：《医学临床研究》2005年第2期145-148,共4页Journal of Clinical Research

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

摘　　要：【目的】探讨热休克反应(HSR)对内毒素(LPS)所致MAPK信号通路的影响。【方法】采用400 ng/mlLPS处理小鼠巨噬细胞(RAW264.7),通过免疫印迹,抗磷酸化抗体检测ERK、JNK、p38的磷酸化。 热休克反应是将巨噬细胞在42℃的条件下培养1h,随后在37℃,5%CO2条件下恢复12h。【结果】ERK、 JNK、p38在LPS处理15min后即被活化,30min达高峰并持续至45min,90min后开始恢复。RT PCR检 测发现TNF α、IL 15mRNA水平增高。细胞经HSR(42℃1h,并恢复12h)后,再经LPS刺激则TNF α、IL 15mRNA转录受抑制,但HSR对JNK、p38、ERK等三种MAPK的活化(磷酸化)无影响。【结论】HSR抑制 LPS所致的炎症因子表达不涉及MAPK信号通路。ObjectiveTo explore the effect of heat shock response(HSR) on the LPS-stimulated MAPK pathways.RAW264.7 murine macrophages were stimulated with lipopolysaccharide(LPS),400 ng/ml. Total RNA was extracted for RT-PCR to detect the expression levels of TNF-α and IL-15 mRNA. Cytoplasmic proteins were extracted and the activation of MAPK pathways was assayed by Western blot using phosphospecific MAPK antibodies.LPS stimulation induced rapid phosphorylatoin of p38, JNK, ERK at 15 min, with a maximal effect at approximately 30 min. These kinases remained phosphorylation at 45 min and almost returned to the baseline levels at 2 h. LPS stimulation increased obviously the mRNA level of TNF-α and IL-15 in macrophages. Heat shock response (42℃1h,recovery for 12 h) inhibited the expression of TNF-α and IL-15, but didn't affect the phosphorylation of MAPK induced by LPS.[Conclusion]The inhibition of heat shock response on the expression of cytokines induced by lipopolysaccharide in murine macrophages does not involve MAPK pathways.

关 键 词：内毒素类 巨噬细胞 炎症 热休克反应 信号传递 小鼠 

分 类 号：R363[医药卫生—病理学]