根皮苷通过调节 LPS 刺激的 RAW264.7 巨噬细胞中的 AMPK/Nrf2/HO-1 信号通路发挥抗炎作用。
Phloroglucinol possesses anti-inflammatory activities by regulating AMPK/Nrf2/HO-1 signaling pathway in LPS-stimulated RAW264.7 murine macrophages.
机构信息
Department of Food and Life Science, Pukyong National University, Busan, Republic of Korea.
Major of Biomedical Engineering, Division of Smart Healthcare, Pukyong National University, Busan, Republic of Korea.
出版信息
Immunopharmacol Immunotoxicol. 2023 Oct;45(5):571-580. doi: 10.1080/08923973.2023.2196602. Epub 2023 Apr 11.
BACKGROUND
Inflammation is closely related to the pathogenesis of chronic illnesses. Secondary metabolites of marine seaweeds are recognized as reliable sources of bioactive compounds due to their health benefits besides their nutritional value. The objective of this study was to determine the potential anti-inflammatory effect of phloroglucinol (Phl) in RAW264.7 murine macrophages after lipopolysaccharides (LPS) stimulation.
METHODS
MTT, nitric oxide (NO), and DCFH-DA assays were conducted to determine cell viability, NO production, and reactive oxygen species (ROS) generation respectively. Pro-inflammatory cytokines and prostaglandin E (PGE) levels were measured using ELISA assay kits. Protein expression levels were determined by western blot analysis.
RESULTS
Phl treatment showed a promising anti-inflammatory effect by reducing NO production, secretion of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6), PGE production, protein expression levels of inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2), and ROS generation in LPS-stimulated RAW264.7 murine macrophages. Phl treatment upregulated heme oxygenase-1 (HO-1) expression by inducing nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) and activating AMPK. However, Zinc protoporphyrin (ZnPP), an inhibitor of HO-1, partially reversed these effects, including NO production, pro-inflammatory cytokine secretion, iNOS, COX-2 and HO-1 expression, and ROS generation.
CONCLUSION
Phl has potential anti-inflammatory activities by regulating AMPK/Nrf2/HO-1 pathway in LPS-stimulated RAW264.7 murine macrophages.
背景
炎症与慢性疾病的发病机制密切相关。海洋藻类的次生代谢产物因其营养价值外的健康益处，被认为是生物活性化合物的可靠来源。本研究旨在确定在脂多糖（LPS）刺激后，间苯三酚（Phl）在 RAW264.7 鼠巨噬细胞中的潜在抗炎作用。
方法
通过 MTT、一氧化氮（NO）和 DCFH-DA 测定法分别测定细胞活力、NO 产生和活性氧（ROS）生成。使用 ELISA 试剂盒测定促炎细胞因子和前列腺素 E（PGE）水平。通过 Western blot 分析测定蛋白表达水平。
结果
Phl 处理通过减少 LPS 刺激的 RAW264.7 鼠巨噬细胞中 NO 产生、促炎细胞因子（TNF-α、IL-1β 和 IL-6）、PGE 产生、诱导型一氧化氮合酶（iNOS）和环加氧酶-2（COX-2）的蛋白表达水平以及 ROS 生成，显示出有希望的抗炎作用。Phl 处理通过诱导核因子红细胞 2 相关因子 2（Nrf2）的核转位和激活 AMPK 而上调血红素加氧酶-1（HO-1）的表达。然而，HO-1 的抑制剂锌原卟啉（ZnPP）部分逆转了这些作用，包括 NO 产生、促炎细胞因子分泌、iNOS、COX-2 和 HO-1 表达以及 ROS 生成。
结论
Phl 通过调节 LPS 刺激的 RAW264.7 鼠巨噬细胞中的 AMPK/Nrf2/HO-1 途径具有潜在的抗炎活性。
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