Daidzein phosphorylates and activates 5-lipoxygenase via the MEK/ERK pathway: a mechanism for inducing the production of 5-lipoxygenase metabolite that inhibit influenza virus intracellular replication
Daidzein phosphorylates and activates 5-lipoxygenase via the MEK/ERK pathway: a mechanism for inducing the production of 5-lipoxygenase metabolite that inhibit influenza virus intracellular replication
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DOI：
10.1016/j.jnutbio.2023.109276
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发表时间：
2023-02-08
影响因子：
5.6
通讯作者：
Shichiri, Mototada
中科院分区：
文献类型：
--
作者：
Horio, Yuka;Isegawa, Yuji;Shichiri, Mototada
我们以前报道过大豆异黄酮（Dz）抑制流感病毒的细胞内复制，花生四烯酸衍生的氧化产物通过脂质氧化酶5-脂氧合酶（5-LOX）参与其抗病毒作用。5-LOX被Dz激活触发抗流感活性;然而，5-LOX激活的机制仍不清楚。因此，在这项研究中，我们的目的是澄清激活机制，使用人单核细胞衍生的THP-1细胞分化佛波醇12-肉豆蔻酸酯13-乙酸酯。THP-1细胞内源性表达5-LOX，Dz不诱导5-LOX表达。然而，用Dz处理8小时后，5-羟基二十碳四烯酸（5-HETE）（通过5-LOX的花生四烯酸氧化产物）的量显著增加，表明酶被激活而不管5-LOX蛋白水平的变化。细胞内Ca2+含量、ATP浓度、5-LOX蛋白磷酸化和5-LOX细胞内定位是已知的5-LOX激活因子。细胞内Ca~（2+）和ATP浓度不受Dz处理的影响。5-LOX的酶活性受三个丝氨酸残基和四个酪氨酸残基的磷酸化调节。用每个激酶的抑制剂预处理显示Dz诱导的5-HETE产生被MEK/ERK抑制剂抑制。5-LOX中的Ser663残基被磷酸化，被发现在Dz处理的THP-1细胞的核组分中增加。免疫细胞化学结果显示，Dz处理后5-LOX易位到核膜。这些结果表明Dz通过MEK/ERK途径磷酸化Ser663激活5-LOX。因此，这些结果表明Dz通过MEK/ERK信号转导途径发挥抗流感病毒活性。(c)2023爱思唯尔公司All rights reserved.
We previously reported that the soy isoflavone daidzein (Dz) suppresses the intracellular replication of influenza virus and that arachidonic acid-derived oxidation product via lipid oxidase 5-lipoxygenase (5-LOX) is involved in its antiviral effect. The activation of 5-LOX by Dz triggers anti-influenza activity; however, the mechanism of activation of 5-LOX remains unclear. Therefore, in this study, we aimed to clarify the activation mechanism using human monocyte-derived THP-1 cells differentiated using phorbol 12-myristate 13-acetate. THP-1 cells expressed 5-LOX endogenously and Dz did not induce 5-LOX expression. However, 8 h after treatment with Dz, the amount of 5-hydroxyeicosatetraenoic acid (5-HETE), an arachidonic acid oxidation product via 5-LOX, increased significantly suggesting that the enzyme is activated regardless of changes in 5-LOX protein levels. Intracellular Ca2+ content, ATP concentration, 5-LOX protein phosphorylation, and 5-LOX intracellular localization are known 5-LOX activation factors. The intracellular Ca2+ and ATP concentrations were not affected by Dz treatment. The enzymatic activity of 5-LOX is regulated by the phosphorylation of three serine residues and four tyrosine residues. Pretreatment with inhibitors of each kinase revealed that Dz-induced 5-HETE production was suppressed by the MEK/ERK inhibitor. 5-LOX in which the Ser663 residue was phosphorylated was found to be increased in the nuclear fraction of Dz-treated THP-1 cells. Furthermore, immunocytochemistry showed that 5-LOX translocates to the nuclear envelope following Dz treatment. These results indicate that Dz activates 5-LOX by phosphorylating Ser663 via the MEK/ERK pathway. Thus, these results demonstrate that Dz exerts anti-influenza virus activity via the MEK/ERK signal transduction pathway. (c) 2023 Elsevier Inc. All rights reserved.