Stimulus (polyphenol, IFN-γ, LPS)-dependent nitric oxide production and antileishmanial effects in RAW 264.7 macrophages
Stimulus (polyphenol, IFN-γ, LPS)-dependent nitric oxide production and antileishmanial effects in RAW 264.7 macrophages
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DOI：
10.1016/j.phytochem.2007.11.012
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发表时间：
2008-12-01
期刊：
影响因子：
3.8
通讯作者：
Kiderlen, Albrecht F.
中科院分区：
文献类型：
--
作者：
Kolodziej, Herbert;Radtke, Oliver A.;Kiderlen, Albrecht F.
本文研究了干扰素、脂多糖和一些多酚作为单独刺激以及不同组合对未感染和感染巨噬细胞样RAW-264.7细胞产生NO的影响，并着重探讨了NO/寄生虫的杀伤关系。在未感染和寄生利什曼原虫的细胞中，没食子酸显著抑制培养上清液中检测到的干扰素-γ和脂多糖诱导的NO。这一效应在干扰素-γ中没有在脂多糖刺激的细胞中那么明显。有趣的是，与未感染的细胞相比，没食子酸只有在干扰素-伽马+脂多糖后3小时内加入才能抑制NO的产生。加入没食子酸不再抑制，有时甚至促进NO的释放。值得注意的是，从所有实验中都明显看出了良好的NO/寄生虫杀灭关系。这项研究扩展到一系列具有免疫刺激活性的多酚(3-O-莽草酸、其3，5-二烯丙基类似物、儿茶素、EGCG和原花青素六聚体)。虽然这些化合物本身被发现是微弱的NO诱导剂，但细胞内利什曼原虫的生存能力大大降低。此外，在干扰素-γ和/或内毒素存在的情况下，测定了它们对巨噬细胞释放NO的剂量依赖效应。同样，未感染细胞和感染细胞对NO的反应有显著差异，而抑制干扰素-γ和/或脂多糖诱导的NO产生强烈地依赖于给定的暴露时间和免疫刺激的顺序。NO水平和利什曼原虫细胞内存活率之间的强烈负相关支持了这样的假设，即观察到的NO抑制不仅仅是由于干扰了用于检测的Griess试验。(C)2007爱思唯尔有限公司。保留所有权利。
The effects of interferon (IFN-gamma), lipopolysaccha ride (LPS), and some polyphenols as individual Stimuli, as well as in various combinations on NO production in non-infected and infected macrophage-like RAW 264.7 cells were investigated, with emphasis on the NO/parasite kill relationship. In non-infected and in Leishmania parasitized cells, gallic acid significantly inhibited the IFN-gamma and LPS-induced NO detected in the supernatant. This effect was less prominent in IFN-gamma- than in LPS-stiniulated cells. Interestingly, and in contrast to non-infected cells, gallic acid inhibited NO production only when added within 3 h after IFN-gamma + LPS. Addition of gallic acid following prolonged incubation with IFN-gamma + LPS periods (24 h) no longer inhibited, sometimes even enhanced NO release. Notably, an excellent NO/parasite kill relationship was evident from all the experiments. This study was extended to a series of polyphenols (3-O-shikimic acid, its 3,5-digalloylated analogue, catechin, EGCG, and a procyanidin hexamer) with proven immunostimulatory activities. Although these compounds themselves were found to be weak NO-inducers, the viability of intracellular Leishmania parasites was considerably reduced. Furthermore, their dose-dependent effects on macrophage NO release was determined in the presence of IFN-gamma and/or LPS. Again, non-infected and infected cells differed significantly in the NO response, while inhibition of IFN-gamma and/or LPS-induced NO production by the tested polyphenols strongly depended on the given time Of exposure and the sequence of immunological stimuli. A strong inverse correlation between NO levels and intracellular survival rates of Leishmania parasites Supported the assumption that the observed inhibition of NO was not simply due to interference with the Griess assay used for detection. (c) 2007 Elsevier Ltd. All rights reserved.