甲基丁香酚：麦卢卡蜂蜜能够改善嗜中性粒细胞内活性氧自由基和 NETosis 的主要驱动因素。

Methyl Syringate: A Primary Driving Factor in Manuka Honeys Ability to Ameliorate Neutrophil Intracellular ROS Activity and NETosis.

机构信息

Department of Biomedical Engineering, University of Memphis, Memphis, TN 38152, USA.

出版信息

Front Biosci (Landmark Ed). 2024 Jul 19;29(7):255. doi: 10.31083/j.fbl2907255.

DOI:10.31083/j.fbl2907255

PMID:39082351

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11973827/

Abstract

BACKGROUND

Neutrophils use both the production of reactive oxygen species (ROS) and a specialized process called NETosis to defend the body from material deemed foreign. While these neutrophil behaviors are critical in preventing infection, a dysregulated response can lead to tissue damage and fibrosis at host-biomaterial interfaces. It was hypothesized that applying the flavonoids found in Manuka honey: chrysin, pinocembrin, and pinobanksin, and the phenolic compound methyl syringate to neutrophils exhibiting pro-inflammatory behavior will reduce ROS activity and prevent NETosis in primary human neutrophils.

METHODS

Using primary human neutrophils isolated from donor (n = 5) peripheral blood, concentrations between 1 nM and 10 µM of each flavonoid, 10 µM and 2 mM of methyl syringate, 0.1% v/v and 10% v/v Manuka honey, and combinations of both 1 nM-10 µM of each flavonoid and 10 µM-2 mM of methyl syringate were assayed for reductions in NETosis using Sytox orange extracellular DNA staining and reduction in intracellular ROS activity via standard dichloro-dihydro-fluorescein diacetate (DCFH-DA) oxidation assay.

RESULTS

Compared to positive control levels, individual flavonoids showed moderate effect sizes. Higher concentrations of flavonoids, especially in combination, stimulated ROS activity by up to 105%. Whole Manuka honey reduced neutrophil extracellular trap (NET) levels by up to 91% but only reduced ROS activity by 36%. However, methyl syringate reduced NET levels by up to 68% and ROS activity by 66%.

CONCLUSIONS

Methyl syringate and whole Manuka honey are potent inhibitors of neutrophil intracellular ROS activity and NET formation. Methyl syringate potentially drives the anti-inflammatory capabilities of Manuka honey demonstrated by previous studies.

摘要

背景

中性粒细胞利用活性氧物质（ROS）的产生和一种称为 NETosis 的专门过程来保护身体免受被视为外来的物质的侵害。虽然这些中性粒细胞的行为对于预防感染至关重要，但失调的反应会导致宿主-生物材料界面的组织损伤和纤维化。据推测，在表现出促炎行为的中性粒细胞上施加在麦卢卡蜂蜜中发现的类黄酮：白杨素、松属素和 pinobanksin 以及酚类化合物甲基丁香酯，将降低 ROS 活性并防止原代人中性粒细胞的 NETosis。

方法

使用从供体（n = 5）外周血分离的原代人中性粒细胞，在 1 nM 至 10 µM 的每种黄酮类化合物、10 µM 和 2 mM 的甲基丁香酯、0.1%v/v 和 10%v/v 的麦卢卡蜂蜜以及 1 nM-10 µM 的每种黄酮类化合物和 10 µM-2 mM 的甲基丁香酯的浓度下，使用 Sytox orange 细胞外 DNA 染色来测定 NETosis 的减少，并通过标准二氯二氢荧光素二乙酸酯（DCFH-DA）氧化测定来测定细胞内 ROS 活性的减少。

结果

与阳性对照水平相比，单个黄酮类化合物表现出中等大小的效应。黄酮类化合物的浓度较高，尤其是在组合中，ROS 活性最高增加 105%。全麦卢卡蜂蜜将中性粒细胞细胞外陷阱（NET）水平降低了高达 91%，但仅将 ROS 活性降低了 36%。然而，甲基丁香酯将 NET 水平降低了高达 68%，ROS 活性降低了 66%。