Zymosan induces NADPH oxidase activation in human neutrophils by inducing the phosphorylation of p47phox and the activation of Rac2: Involvement of protein tyrosine kinases, PI3Kinase, PKC, ERK1/2 and p38MAPkinase
Zymosan induces NADPH oxidase activation in human neutrophils by inducing the phosphorylation of p47phox and the activation of Rac2: Involvement of protein tyrosine kinases, PI3Kinase, PKC, ERK1/2 and p38MAPkinase
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DOI：
10.1016/j.bcp.2012.10.010
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发表时间：
2013-01-01
影响因子：
5.8
通讯作者：
El-Benna, Jamel
中科院分区：
文献类型：
--
作者：
Makni-Maalej, Karama;Chiandotto, Melanie;El-Benna, Jamel
由中性粒细胞NADPH氧化酶产生的活性氧（ROS）在宿主防御病原体（如细菌和真菌）中起着关键作用。酵母菌（Zymosan）是一种从酿酒酵母中提取的细胞壁制剂，主要用于激活中性粒细胞的调理形式。在这项研究中，我们发现非调理酶酶体单独诱导人中性粒细胞产生ROS。酵素诱导的ROS产量高于甲酰基蛋氨酸-亮氨酸-苯丙氨酸（fMLF）或肉豆酸酯（PMA）诱导的ROS产量，但低于琼脂化酵素诱导的ROS产量。大多数酵素诱导的ROS产生是在细胞内。有趣的是，zymosan诱导NADPH氧化酶胞质组分p47phox在Ser315， Ser328和Ser345位点磷酸化。Zymosan还诱导了小g蛋白Rac2的活化。染料木黄酮（染料木黄酮是一种广谱蛋白酪氨酸激酶抑制剂）和wortmannin（一种pi3k激酶抑制剂）抑制了p47phox的磷酸化和Rac2的激活。GF109203X PKC抑制剂抑制p47phox在Ser315和Ser328上的磷酸化。p38MAPK和erk1 /2通路抑制剂SB203580和UO126分别抑制p47phox在Ser345上的磷酸化。染料木素和wortmannin完全抑制zymosan诱导的ROS生成，SB203580、UO126和GF109203X部分抑制zymosan诱导的ROS生成。这些结果表明，单独的zymosan能够通过磷酸化p47phox和激活Rac2激活人中性粒细胞中的NADPH氧化酶，并且蛋白酪氨酸激酶、PI3Kinase、p38MAPK、ERK1/2和PKC参与了这一过程。这些途径可能是治疗酵母菌和酿酒葡萄球菌引起的炎症的潜在药理靶点。(c) 2012 Elsevier Inc.版权所有。
Reactive oxygen species (ROS) production by the neutrophil NADPH oxidase plays a key role in host defense against pathogens, such as bacteria and fungi. Zymosan a cell-wall preparation from Saccharomyces cerevisiae is largely used to activate neutrophils in its opsonized form. In this study, we show that non-opsonized zymosan alone induced ROS production by human neutrophils. Zymosan-induced ROS production is higher than the formyl-methionyl-leucyl-phenylalanine (fMLF)- or the phorbol myristate acetate (PMA)-induced ROS production but is lower than the one induced by opsonized zymosan. Most of the zymosan-induced ROS production is intracellular. Interestingly, zymosan induced the phosphorylation of the NADPH oxidase cytosolic component p47phox on several sites which are Ser315, Ser328 and Ser345. Zymosan induced also the activation of the small G-protein Rac2. Phosphorylation of the p47phox as well as Rac2 activation were inhibited by genistein a broad range protein tyrosine kinase inhibitor and by wortmannin a PI3Kinase inhibitor. GF109203X a PKC inhibitor inhibited phosphorylation of p47phox on Ser315 and Ser328. SB203580 and UO126, inhibitors of p38MAPK and ERK1/2-pathway, respectively, inhibited phosphorylation of p47phox on Ser345. Zymosan-induced ROS production was completely inhibited by genistein and wortmannin and partially inhibited by SB203580, UO126 and GF109203X. These results show that zymosan alone is able to activate NADPH oxidase in human neutrophils via the phosphorylation of p47phox and Rac2 activation and that a protein tyrosine kinase, PI3Kinase, p38MAPK, ERK1/2 and PKC are involved in this process. These pathways could be potential pharmacological targets to treat zymosan- and S. cerevisiae-induced inflammation. (c) 2012 Elsevier Inc. All rights reserved.