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

发布时间：2018-02-26 07:03

本文关键词： 过氧化氢肿瘤坏死因子-α 白细胞介素-β 单核细胞趋化蛋白- 巨噬细胞炎症蛋白- 巨噬细胞　出处：《中国生物化学与分子生物学报》2015年07期 　论文类型：期刊论文

【摘要】：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.
[Abstract]:Reactive oxygen species (Ros) produced by immunoreactive cells induced by respiratory waterfall are signaling molecules of macrophage expression of pro-inflammatory cytokines and chemokines. However, there is no direct evidence that H _ 2O _ 2 can stimulate the expression of pro-inflammatory cytokines and chemokines in macrophages. In this study, mouse RAW264.7 macrophages cultured in vitro were used as the study system. Effects of exogenous H _ 2O _ 2 on expression and production of proinflammatory and chemokine genes in RAW264.7 macrophages. MTT assay combined with real-time fluorescence quantification of PCRQ RT-PCRN. Elisa results showed that RAW264.7 cells had no effect on RAW264.7 fineness when H _ 2O _ 2 concentration was below 40 渭 mol/L. Cell proliferation activity. 20 渭 mol/L and 40 渭 mol/L H2O2 significantly enhanced the transcription and protein production of MCP-1 and MIP-2 genes in RAW264.7 cells. And there was a dose-dependent effect. However, pretreatment with 200UmL catalase could attenuate the expression and protein production of MCP-1 and MIP-2 genes stimulated by H2O2. These results suggest that H2O2 is an important factor in stimulating the expression or production of inflammatory and chemokines in macrophages. It plays an important role in the occurrence of inflammatory reaction.
【作者单位】：河南师范大学生命科学学院;
【基金】：河南省重点科技攻关计划(No.122102310282) 河南省教育厅自然科学基础研究计划(No.2010A180011)~~
【分类号】：R363

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