2-甲氧基-4-乙烯基苯酚的抗炎活性涉及通过血红素加氧酶-1 抑制脂多糖诱导的诱导型一氧化氮合酶。

Anti-inflammatory activity of 2-methoxy-4-vinylphenol involves inhibition of lipopolysaccharide-induced inducible nitric oxidase synthase by heme oxygenase-1.

机构信息

Division of Dental Pharmacology, Faculty of Dentistry and Graduate School of Medical and Dental Science, Niigata University, Niigata, Japan.

Division of Reconstructive Surgery for Oral and Maxillofacial Region, Faculty of Dentistry and Graduate School of Medical and Dental Science, Niigata University, Niigata, Japan.

出版信息

Immunopharmacol Immunotoxicol. 2023 Oct;45(5):589-596. doi: 10.1080/08923973.2023.2197141. Epub 2023 Apr 5.

DOI:10.1080/08923973.2023.2197141

PMID:36995736

Abstract

BACKGROUND

2-Methoxy-4-vinylphenol (2M4VP) is a natural anti-inflammatory compound derived from red wine, but its underlying mechanism remains unclear. Heme oxygenase-1 (HO-1), an anti-inflammatory enzyme, inhibits gene expression, while nuclear factor erythroid 2-related factor 2 (Nrf2), a transcription factor involved in HO-1 production, binds to the antioxidant response element (ARE) in the nucleus and promotes HO-1 transcription. Based on the hypothesis that the inhibitory effect of 2M4VP on NO production is mediated by HO-1, we examined the possible mechanism of the anti-inflammatory activity of 2M4VP in this study.

MATERIALS AND METHODS

The anti-inflammatory activity of 2M4VP was analyzed by Griess method, ELISA, qPCR, and Western blotting using LPS-treated macrophage lineage RAW264.7 cells. The impact of 2M4VP on the Nrf2/ARE pathway was also analyzed using immunocytochemistry and an ARE luciferase reporter using HEK293 cells.

RESULTS

The results showed that 2M4VP reduced the production of LPS-induced NO and inducible nitric oxidase synthase (iNOS). In addition, 2M4VP increased the expression of HO-1, while pretreatment with the Nrf2 inhibitor ML385 downregulated HO-1 expression. 2M4VP induced Kelch-like ECH-associated protein 1 (Keap1) degradation. Furthermore, it promoted Nrf2 nuclear translocation and increased luciferase activity by binding to the ARE.

CONCLUSIONS

2M4VP induces Keap1 degradation and promotes Nrf2 nuclear translocation. Activation of Nrf2/ARE pathway enhances HO-1 expression and leads to iNOS inhibition for anti-inflammatory function.

摘要

背景

2-甲氧基-4-乙烯基苯酚（2M4VP）是一种天然抗炎化合物，来源于红酒，但它的作用机制尚不清楚。血红素加氧酶-1（HO-1）是一种抗炎酶，可抑制基因表达，而核因子红细胞 2 相关因子 2（Nrf2）作为一种参与 HO-1 生成的转录因子，可与核内抗氧化反应元件（ARE）结合，并促进 HO-1 转录。基于 2M4VP 抑制 NO 产生的作用是通过 HO-1 介导的假说，我们在本研究中检测了 2M4VP 抗炎活性的可能机制。

材料和方法

采用 Griess 法、ELISA、qPCR 和 Western blot 分析 LPS 处理的巨噬细胞系 RAW264.7 细胞中 2M4VP 的抗炎活性。还采用免疫细胞化学和 HEK293 细胞中的 ARE 荧光素酶报告基因分析 2M4VP 对 Nrf2/ARE 通路的影响。

结果

结果表明，2M4VP 降低了 LPS 诱导的 NO 和诱导型一氧化氮合酶（iNOS）的产生。此外，2M4VP 增加了 HO-1 的表达，而 Nrf2 抑制剂 ML385 的预处理下调了 HO-1 的表达。2M4VP 诱导 Kelch-like ECH-associated protein 1（Keap1）降解。此外，它通过与 ARE 结合促进 Nrf2 核易位并增加荧光素酶活性。