5-脂氧合酶依赖性生物合成具有抗炎活性的新型 20:4n-3 代谢物。

5-lipoxygenase-dependent biosynthesis of novel 20:4 n-3 metabolites with anti-inflammatory activity.

机构信息

Départment de Chimie et Biochimie, Université de Moncton, 18 Antonine-Maillet Avenue, Moncton, NB E1A 3E9, Canada.

Départment de Chimie et Biochimie, Université de Moncton, 18 Antonine-Maillet Avenue, Moncton, NB E1A 3E9, Canada; Centre de recherche de l'Institut universitaire de cardiologie et de pneumologie de Québec, Université Laval, Québec, QC, Canada.

出版信息

Prostaglandins Leukot Essent Fatty Acids. 2018 Nov;138:38-44. doi: 10.1016/j.plefa.2018.10.005. Epub 2018 Oct 16.

DOI:10.1016/j.plefa.2018.10.005

PMID:30392579

Abstract

5-lipoxygenase (5-LO) catalyzes the conversion of arachidonic acid (AA) into pro-inflammatory leukotrienes. N-3 PUFA like eicosapentaenoic acid are subject to a similar metabolism and are precursors of pro-resolving mediators. Stearidonic acid (18:4 n-3, SDA) is a plant source of n-3 PUFA that is elongated to 20:4 n-3, an analogue of AA. However, no 5-LO metabolites of 20:4 n-3 have been reported. In this study, control and 5-LO-expressing HEK293 cells were stimulated in the presence of 20:4 n-3. Metabolites were characterized by LC-MS/MS and their anti-inflammatory properties assessed using AA-induced autocrine neutrophil stimulation and leukotriene B-mediated chemotaxis. 8‑hydroxy‑9,11,14,17-eicosatetraenoic acid (Δ-8-HETE) and 8,15-dihydroxy-9,11,13,17-eicosatetraenoic acid (Δ-8,15-diHETE) were identified as novel metabolites. Δ-8,15-diHETE production was inhibited by the leukotriene A hydrolase inhibitor SC 57461A. Autocrine neutrophil leukotriene stimulation and neutrophil chemotaxis, both BLT1-dependent processes, were inhibited by Δ-8,15-diHETE at low nM concentrations. These data support an anti-inflammatory role for Δ-8,15-diHETE, a novel 5-LO product.

摘要

5-脂氧合酶（5-LO）催化花生四烯酸（AA）转化为促炎白三烯。像二十碳五烯酸（EPA）这样的 n-3PUFA 也经历类似的代谢，并作为促分解介质的前体。硬脂酸（18:4n-3，SDA）是一种植物来源的 n-3PUFA，可被延伸为 20:4n-3，AA 的类似物。然而，尚未报道 20:4n-3 的 5-LO 代谢物。在这项研究中，在存在 20:4n-3 的情况下，用对照和 5-LO 表达的 HEK293 细胞进行刺激。通过 LC-MS/MS 对代谢产物进行了表征，并使用 AA 诱导的自分泌中性粒细胞刺激和白三烯 B 介导的趋化作用评估了它们的抗炎特性。鉴定出 8-羟基-9,11,14,17-二十碳四烯酸（Δ-8-HETE）和 8,15-二羟基-9,11,13,17-二十碳四烯酸（Δ-8,15-二 HETE）为新型代谢产物。白三烯 A 水解酶抑制剂 SC 57461A 抑制了 Δ-8,15-二 HETE 的产生。自分泌中性粒细胞白三烯刺激和中性粒细胞趋化作用均为 BLT1 依赖性过程，在低 nM 浓度下被 Δ-8,15-二 HETE 抑制。这些数据支持 Δ-8,15-二 HETE，一种新型 5-LO 产物的抗炎作用。