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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.
Reactive oxygen species produced by respiratory cascades are signaling molecules for the expression of proinflammatory cytokines and chemokines in macrophages. However, the current lack of hydrogen peroxide (H2O2) stimulates macrophages to express pro-inflammatory cytokines and chemokines Of the direct evidence.In this study, ex vivo cultured mouse RAW264.7 macrophages as a research system to explore the exogenous H2O2 on RAW264.7 macrophages proinflammatory cytokines and chemokines gene expression and production.MTT method Combined with real-time quantitative PCR (qRT-PCR), the results of enzyme-linked immunosorbent assay (ELISA) showed that RAW264.7 cells did not affect the viability of RAW264.7 cells when H2O2 concentration was lower than 40μmol / L. / L H2O2 significantly increased the gene transcription and protein production of TNF-α, IL-1β, MCP-1 and MIP-2 in RAW264.7 cells in a dose-dependent manner. However, pretreatment with 200 U / m L catalase Attenuated H2O2-induced TNF-α, IL-1β, MCP-1 and MIP-2 gene expression and protein production.These results suggest that H2O2 is an important factor to stimulate the expression or production of proinflammatory cytokines and chemokines in macrophages, The body’s inflammatory response has an important role.