【摘要】 【目的】观察热休克反应 (HSR)以及热休克转录因子 1(HSF1)对LPS诱导的TNF α和IL 15表达的影响。【方法】用 2 0 0ng/mlLPS处理热休克或未热休克的小鼠RAW 2 6 4 .7巨噬细胞 ,抽提细胞的总RNA进行RT PCR实验 ,观察TNF α和IL 15mRNA表达的情况 ;采用HSF1基因敲除小鼠经腹腔内注射LPS复制内毒素血症模型 ,抽提HSF1基因敲除小鼠 (HSF1-/ -)和野生型小鼠 (HSF1+ / + )肺组织的总RNA进行RT PCR实验 ,观察TNF α和IL 15mRNA表达的情况。用TESS分析IL 15的启动子 ,寻找热休克元件(Heatshockelement,HSE)。【结果】小鼠RAW 2 6 4 .7巨噬细胞受LPS刺激后 ,TNF α和IL 15mRNA的水平明显增加 ,而热休克抑制了LPS诱导的TNF α和IL 15mRNA的表达 ;腹腔注射LPS后 ,HSF1-/ -小鼠和HSF1+ / + 小鼠的肺组织中TNF α和IL 15mRNA的水平明显增加 ,HSF1-/ -小鼠的肺组织中TNF α和IL 15mRNA水平的增加明显高于HSF1+ / + 小鼠。经TESS软件分析发现IL 15的启动子区含有 2个HSE。【结论】HSR、HSF1抑制了LPS诱导的TNF α和IL 15的表达 ;HSF1可能通过与IL 15启动子区的HSE结合抑制LPS诱导的IL 15的表达。更多还原

【Abstract】 [Objectives]To study the effect of heat shock response (HSR) and heat shock transcription factor-1 (HSF-1)on the LPS-induced expression of TNF-α and IL-15.Macrophages collected from heat-shocked and untreated RAW264.7 mice were stimulated with 200ng/ml lipopolysaccharide (LPS).Total RNA was extracted and RT-PCR assay was performed to detect the expression levels of TNF-α mRNA and IL-15mRNA. HSF-1 -/- mice (gene knocked mice) and HSF-1 +/+ mice (wild type mice) were injected with 15mg/kg LPS intraperitoneally(ip) and total RNA of lung tissues was extracted. The mRNA expressions of TNF-α and IL-15. were detected by RT-PCR. The promoter of IL-15 was analyzed by TESS program to search for heat shock element(HSE).LPS stimulation increased obviously the mRNA levels of TNF-α and IL-15 in macrophages of RAW264.7 mice, which was inhibited by heat shock pretreatment. Intraperitoneal injection of LPS induced the accumulation of TNF-α and IL-15 mRNA in the lungs of mice carrying both genotypes, but the increase of TNF-α and IL-15 mRNA level in HSF-1 -/- mice was more prominent than that in HSF-1 +/+ mice. Analysis with TESS software revealed that promoter of IL-15 gene contained two complete HSEs.[Conclusion]HSR or HSF-1 inhibits LPS-induced expression of TNF-α and IL-15 ; HSF-1 may inhibit LPS-induced expression of IL-15 by binding to HSEs in the promoter of IL-15 gene.更多还原