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An Optimized Ex Vivo n–3 PUFA Supplementation Strategy for Primary Human Macrophages Shows That DHA Suppresses Prostaglandin E2 Formation
Molecular Nutrition & Food Research ( IF 4.5 ) Pub Date : 2024-12-16 , DOI: 10.1002/mnfr.202400716
Rebecca Kirchhoff, Nadja Kampschulte, Carina Rothweiler, Nadine Rohwer, Karsten‐Henrich Weylandt, Nils Helge Schebb

Evidence suggests beneficial effects of long‐chain n–3 polyunsaturated fatty acids (PUFAs) in inflammatory diseases. However, the underlying mechanisms are still subject of research. For this purpose, we developed an ex vivo n–3 PUFA supplementation strategy. M2‐like macrophages were supplemented for 2–3 days with 20–40 µM docosahexaenoic acid (DHA) during differentiation. Quality parameters include <3% oxylipins for PUFA‐preparation, total fatty acids (FAs) <10 mM, and low oxylipins in plasma, n–3 PUFA <0.25 mM for the selection of donors of plasma as well as %n–6 in highly unsaturated fatty acids (HUFAs) ≥70% for donors of cells. Following supplementation, PUFA pattern of cells was shifted toward one described for blood and tissue from subjects with higher n–3 and lower n–6 PUFAs. This was accompanied by a decrease of arachidonic acid‐derived oxylipins in a dose‐ and time‐dependent manner in favor of n–3 PUFA ones. Stimulation with LPS resulted in decreased levels of pro‐inflammatory prostaglandins in the DHA‐supplemented cells, but no changes in cytokines. In vitro supplementation studies with n–3 PUFA need rigorous controls to exclude the background formation of oxylipins. By accounting for these possible confounders the described approach allows the mechanistic investigation of n–3 PUFAs in primary human immune cells, offering an alternative for intervention studies.

中文翻译:


针对原代人巨噬细胞的优化体外 n-3 PUFA 补充策略显示,DHA 抑制前列腺素 E2 的形成



有证据表明,长链 n-3 多不饱和脂肪酸 (PUFA) 对炎症性疾病有益。然而,潜在的机制仍然是研究的主题。为此,我们开发了一种离体 n-3 PUFA 补充策略。在分化过程中,向 M2 样巨噬细胞补充 20-40 μM 二十二碳六烯酸 (DHA) 2-3 天。质量参数包括用于 PUFA 制备的 <3% 脂氧化物、总脂肪酸 (FA) <10 mM 和血浆中的低脂氧化物、用于选择血浆供体的 n-3 PUFA <0.25 mM,以及高度不饱和脂肪酸 (HUFA) 中的 %n-6 ≥70% 用于细胞供体。补充后,细胞的 PUFA 模式转变为 n-3 较高和 n-6 PUFA 较低的受试者的血液和组织模式。这伴随着花生四烯酸衍生的脂氧化物以剂量和时间依赖性方式减少,有利于 n-3 个 PUFA 的。用 LPS 刺激导致补充 DHA 的细胞中促炎前列腺素水平降低,但细胞因子没有变化。使用 n-3 PUFA 的体外补充剂研究需要严格的控制,以排除氧磷脂的背景形成。通过考虑这些可能的混杂因素,所描述的方法允许对原代人类免疫细胞中的 n-3 个 PUFA 进行机制研究,为干预研究提供了替代方案。
更新日期:2024-12-16
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