过氧化氢刺激RAW264.7巨噬细胞促炎细胞因子和趋化因子基因表达

免疫反应细胞经呼吸瀑布作用产生的活性氧是巨噬细胞促炎细胞因子和趋化因子表达的信号分子,但目前缺乏过氧化氢(H2O2)刺激巨噬细胞表达促炎细胞因子和趋化因子的直接证据.本研究以离体培养的小鼠RAW264.7巨噬细胞为研究体系,探讨外源H2O2对RAW264.7巨噬细胞促炎因子和趋化因子基因表达和生成的影响.MTT法结合实时荧光定量PCR(qRT-PCR)、酶联免疫吸附试验(ELISA)结果显示,RAW264.7细胞在H2O2浓度低于40μmol/L时不影响RAW264.7细胞的增殖活力.20μmol/L和40μmol/L H2O2显著增强RAW264.7细胞TNF-α、IL-1β、MCP-1和MIP-2基因转录和蛋白质生成,并存在剂量依赖效应;而200 U/m L过氧化氢酶预处理则可减弱由H2O2刺激的TNF-α、IL-1β、MCP-1和MIP-2基因表达和蛋白生成.这些结果提示,H2O2是刺激巨噬细胞促炎因子和趋化因子表达或生成的重要因子,对机体炎症反应的发生具有重要作用.

Hydrogen Peroxide Stimulates Expression of Cytokine and Chemokine Genes in Murine RAW264.7 Cells

Hydrogen peroxide (H2O2), as a major form of reactive oxygen species (ROS), produced from cell respiratory burst functions as signal molecules in promoting the expression of pro-inflammatory cytokine/chemokine genes in macrophage. We investigated H2O2 timulation on the expression of pro-inflammatory cytokine/chemokine genes in the murine RAW264.7 cells. From the results of MTT, with qRT-PCR and ELISA assays, H2O2 was showed to markedly increase Tnf-α, Il-1β, Mcp-1 and Mip-2 gene expression at 20 ～ 40 μmol/L without obvious cytotoxicity. Catalase treatment at 200 U/mL reduced H2O2-stimulated expression of the up-mentioned gene. These data suggested that H2O2 was an important stimulator regulating the expression of pro-inflammatory cytokines/chemokines in macrophages.