The spin trap 5,5-dimethyl-1-pyrroline N-oxide inhibits lipopolysaccharide-induced inflammatory response in RAW 264.7 cells
The spin trap 5,5-dimethyl-1-pyrroline N-oxide inhibits lipopolysaccharide-induced inflammatory response in RAW 264.7 cells
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DOI：
10.1016/j.lfs.2011.12.018
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发表时间：
2012-03-10
期刊：
影响因子：
6.1
通讯作者：
Ramirez, Dario C.
中科院分区：
文献类型：
--
作者：
Zhai, Zili;Gomez-Mejiba, Sandra E.;Ramirez, Dario C.
目的：巨噬细胞暴露于脂多糖（LPS）诱导氧化和炎症应激，导致细胞损伤。抗氧化和抗炎特性已归因于硝酮自旋陷阱5，5-二甲基-1-吡咯啉N-氧化物（DMPO），通常用于自由基分析，但DMPO的这些方面很少探索。在这项研究中，我们试图建立DMPO的抗炎活性，大概是通过清除自由基，否则有助于激活炎症反应和损伤细胞。主要方法：用LPS和/或DMPO处理RAW 264.7巨噬细胞不同的时间点，细胞损伤，炎症介质的产生，诱导型一氧化氮合酶（iNOS）表达，NF-κ B p65活化，关键发现：在细胞用LPS和/或DMPO处理24小时后，DMPO减少LPS诱导的炎症反应，如通过下调iNOS表达和炎症介质的产生所指示的。因此，DMPO保护细胞免受LPS诱导的细胞毒性。为了理解这些DMPO作用的机制基础，检测了NF-κ B p65活化以及MAPK和Akt的磷酸化。我们发现，通过测定用LPS和/或DMPO处理15-60分钟的细胞，DMPO抑制MAPK的磷酸化。Akt和I κ B α，并减少NF-κ B p65易位。意义：DMPO具有抗炎活性，减轻LPS诱导的细胞损伤，其机制可能是通过减少ROS的产生，从而阻止随后的炎症激活和损伤。(C)2012 Elsevier Inc. All rights reserved.
Aim: Exposure of macrophages to lipopolysaccharide (LPS) induces oxidative and inflammatory stresses, which cause cell damage. Antioxidant and anti-inflammatory properties have been attributed to the nitrone spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO), commonly used in free radical analysis, but these aspects of DMPO have been little explored. In this study, we sought to establish the anti-inflammatory activity of DMPO, presumably by removing free radicals which otherwise help activate inflammatory response and damage cells.Main methods: RAW 264.7 macrophages were treated with LPS and/or DMPO for different time points, cell damage, production of inflammatory mediators, inducible nitric oxide synthase (iNOS) expression, NF-kappa B p65 activation, phosphorylation of MAPKs and Akt, and intracellular reactive oxygen species (ROS) were determined.Key findings: After cells were treated with LPS and/or DMPO for 24 h, DMPO reduced the LPS-induced inflammatory response as indicated by downregulated iNOS expression and production of inflammatory mediators. Accordingly, DMPO protected cells from LPS-induced cytotoxicity. In order to understand the mechanistic basis of these DMPO effects, the NF-kappa B p65 activation and the phosphorylation of MAPKs and Akt were examined. We found, by assaying cells treated with LPS and/or DMPO for 15-60 min, that DMPO inhibited the phosphorylation of MAPKs. Akt, and I kappa B alpha, and reduced the NF-kappa B p65 translocation. Furthermore, we demonstrated that DMPO inhibited LPS-induced ROS production.Significance: DMPO showed the anti-inflammatory activity and attenuated LPS-induced cell damage, most likely by reducing ROS production and thus preventing the subsequent inflammatory activation and damage. (C) 2012 Elsevier Inc. All rights reserved.