Eugenol prevents fMLF-induced superoxide anion production in human neutrophils by inhibiting ERK1/2 signaling pathway and p47phox phosphorylation
Eugenol prevents fMLF-induced superoxide anion production in human neutrophils by inhibiting ERK1/2 signaling pathway and p47phox phosphorylation
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DOI：
10.1038/s41598-019-55043-8
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发表时间：
2019-12-06
影响因子：
4.6
通讯作者：
Bachoual, Rafik
中科院分区：
文献类型：
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作者：
Chniguir, Amina;Pintard, Coralie;Bachoual, Rafik
丁香酚是从丁香精油中提取的多酚。已知它具有抗炎和化学保护特性，以及由于其酚基的存在而具有有效的抗氧化活性。在这项研究中，我们研究了丁香酚对中性粒细胞超氧化物生成的影响，这是先天免疫和炎症的关键过程。用细胞色素c还原法测定人中性粒细胞超氧阴离子的产生。Western blotting分析NADPH氧化酶激活的关键蛋白p47 phox、MAPK激酶（p38和ERK 1/2）、MEK 1/2和Raf的磷酸化。通过增加浓度（2.5 μ g/mL-20 μ g/mL）的丁香酚预处理嗜中性粒细胞30分钟，显著抑制（p < 0.001）由趋化肽甲酰-Met-Leu-Phe（fMLF）诱导的超氧阴离子产生，IC 50为5 μ g/mL。佛波肉豆蔻酸酯乙酸酯（PMA）刺激的O2（-）生产的影响，只有在最高的丁香酚浓度（20微克/毫升）。结果表明，丁香酚可抑制p47 phox在Ser-345和Ser-328上的磷酸化，抑制p47 phox向细胞膜的转位，抑制Raf、MEK 1/2和ERK 1/2蛋白的磷酸化。综上所述，我们的研究结果表明，丁香酚通过抑制Raf/MEK/ERK 1/2/p47 phox磷酸化途径抑制中性粒细胞产生超氧阴离子。
Eugenol is a polyphenol extracted from Syzygium aromaticum essential oil. It is known to have anti-inflammatory and chemoprotective properties as well as a potent anti-oxidant activity due the presence of its phenolic group. In this study, we examined the effects of eugenol on neutrophil superoxide production, a key process involved in innate immunity and inflammation. Superoxide anion generationin human neutrophils was measured by cytochrome c reduction assay. Western blotting was used to analyze the phosphorylation of, p47phox, MAPKinases (p38 and ERK1/2), MEK1/2 and Raf, key proteins involved in the activation of NADPH oxidase. Pretreatment of neutrophils by increasing concentrations (2.5 mu g/mL-20 mu g/mL) of eugenol for 30 min, inhibited significantly (p < 0.001) superoxide anion generation induced by the chemotactic peptide formyl-Met-Leu-Phe (fMLF) with an IC50 of 5 mu g/mL. Phorbolmyristate acetate (PMA)-stimulated O2(-) production was affected only at the highest eugenol concentration (20 mu g/mL). Results showed that eugenol decreased the phosphorylation of p47phox onSer-345 and Ser-328, the translocation of p47phox to the membranesand the phosphorylation of Raf, MEK1/2 and ERK1/2 proteins. Taken together, our results suggest that eugenol inhibits the generation of superoxide anion by neutrophils via the inhibition of Raf/MEK/ERK1/2/p47phox-phosphorylation pathway.