Hydrogen peroxide induces murine macrophage chemokine gene transcription via extracellular signal-regulated kinaseand cyclic adenosine 5′-monophosphate (cAMP)-dependent pathways:: Involvement of NF-κB, activator protein 1, and cAMP response element binding protein
Hydrogen peroxide induces murine macrophage chemokine gene transcription via extracellular signal-regulated kinaseand cyclic adenosine 5′-monophosphate (cAMP)-dependent pathways:: Involvement of NF-κB, activator protein 1, and cAMP response element binding protein
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DOI：
10.4049/jimmunol.169.12.7026
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发表时间：
2002-12-15
影响因子：
4.4
通讯作者：
Olivier, M
中科院分区：
文献类型：
--
作者：
Jaramillo, M;Olivier, M
过氧化氢 (H2O2) 已被证明可作为激活趋化因子表达的第二信使。在本研究中，我们研究了小鼠巨噬细胞系 B10R 中这种细胞调节的机制。我们报告，H2O2 通过激活细胞外信号调节激酶 (ERK) 途径和 转录因子 NF-kappaB、AP-1 和 CREB 的核转位。使用丝裂原激活蛋白激酶激酶 1/2 和 ERK1/ERK2 的特异性抑制剂阻断 ERK 通路，完全消除了 H2O2 介导的趋化因子上调和所有研究的 NF 的激活。同样，选择性抑制 cAMP 和 NF-kappaB 分别强烈下调所有趋化因子转录物的诱导以及 CREB 和 NF-kappaB 的激活。有趣的是，当添加特定的冷寡核苷酸（NF-kappaB、AP-1 和 CREB）或针对各种转录因子亚基（p50、p65、c-Fos、Jun B、c-Jun 和 CREB-1）的抗体时，我们检测到 NF-kappaB、AP-1 和 CREB DNA 结合活性显着降低，因为这些结合位点相互竞争。这些发现表明，ERK 和 cAMP 依赖性途径之间的合作似乎是实现最大 H2O2 依赖性趋化因子调节的必需转录因子复合物的形成所必需的。最后，用放线菌素 D 进行的实验表明，H2O2 介导的 MIP-1beta mRNA 上调是转录控制的结果，而 MIP-1α、MIP-2 和单核细胞趋化蛋白-1 的上调是由于基因转录激活和 mRNA 转录后稳定所致。
Hydrogen peroxide (H2O2) has been shown to act as a second messenger that activates chemokine expression. In the present study, we investigated the mechanisms underlying this cellular regulation in the murine macrophage cell line B10R. We report that H2O2 increases mRNA expression of various chemokines, macrophage-inflammatory protein (MIP)-1alpha/CC chemokine ligand (CCL)3, MIP-1beta/CCL4, MIP-2/CXC chemokine ligand 2, and monocyte chemoattractant protein-1/CCL2, by activating the extracellular signal-regulated kinase (ERK) pathway and the nuclear translocation of the transcription factors NF-kappaB, AP-1, and CREB. Blockage of the ERK pathway with specific inhibitors against mitogen-activated protein kinase kinase 1/2 and ERK1/ERK2 completely abolished both the H2O2-mediated chemokine up-regulation and the activation of all NF studied. Similarly, selective inhibition of cAMP and NF-kappaB strongly down-regulated the induction of all chemokine transcripts as well as CREB and NF-kappaB activation, respectively. Of interest, we detected a significant decrease of NF-kappaB, AP-1, and CREB DNA binding activities by reciprocal competition for these binding sites when either specific cold oligonucleotides (NF-kappaB, AP-1, and CREB) or Abs against various transcription factor subunits (p50, p65, c-Fos, Jun B, c-Jun, and CREB-1) were added. These findings indicate that cooperation between ERK- and cAMP-dependent pathways seems to be required to achieve the formation of an essential transcriptional factor complex for maximal H2O2-dependent chemokine modulation. Finally, experiments performed with actinomycin D suggest that H2O2-mediated MIP-1beta mRNA up-regulation results from transcriptional control, whereas that of MIP-1alpha, MIP-2, and monocyte chemoattractant protein-1 is due to both gene transcription activation and mRNA posttranscriptional stabilization.