Background: The macrophage is one of the most important types of immune cells that protect against harmful stimuli. Macrophage activation plays a pivotal role in the progression and development of various inflammatory diseases. The neurokinin 1 receptor (NK-1R) is a G protein-coupled receptor that plays an important role in inflammatory diseases. Aprepitant is a kind of NK-1R antagonist. The purpose of this study is to determine the protective effect of aprepitant in lipopolysaccharide (LPS)-induced inflammatory responses in macrophages.
Methods: We examined the anti-inflammatory and anti-oxidant effects of aprepitant in LPS-treated RAW264.7 macrophages by using real-time PCR, ELISA, and Western blot analysis. We also assessed cellular oxidative stress signaling by measuring the levels of cellular MDA, total ROS, and NADPH oxidase expression. Cellular NO production was measured by DAF-FM DA staining. The inhibitory effect of aprepitant against NF-κB signaling was evaluated by luciferase assay and Western blot analysis.
Results: The expression of NK-1R is increased in LPS-induced macrophages, suggesting a potential role of the receptor in the inflammatory response. We show that aprepitant protects macrophages against oxidative stress by reducing the generation of ROS and the expression of NOX-4. Furthermore, aprepitant inhibits the secretion of pro-inflammatory cytokines and chemotactic factors by mediating the NF-κB signaling pathway.
Conclusion: The NK-1R receptor antagonist aprepitant acts as an anti-inflammatory agent, indicating that the blockage of the NK-1R pathway in macrophages has the potential to suppress inflammation.



中文翻译：

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NK-1R 拮抗剂 Aprepitant 可预防 LPS 诱导的 RAW264.7 巨噬细胞的氧化应激和炎症。

背景：巨噬细胞是抵御有害刺激的最重要的免疫细胞类型之一。巨噬细胞活化在各种炎症性疾病的进展和发展中起关键作用。神经激肽 1 受体 (NK-1R) 是一种 G 蛋白偶联受体，在炎症性疾病中起重要作用。阿瑞匹坦是一种NK-1R拮抗剂。本研究的目的是确定阿瑞匹坦在脂多糖 (LPS) 诱导的巨噬细胞炎症反应中的保护作用。
方法：我们使用实时 PCR、ELISA 和蛋白质印迹分析检测了阿瑞匹坦在 LPS 处理的 RAW264.7 巨噬细胞中的抗炎和抗氧化作用。我们还通过测量细胞 MDA、总 ROS 和 NADPH 氧化酶表达的水平来评估细胞氧化应激信号。通过 DAF-FM DA 染色测量细胞 NO 的产生。通过荧光素酶测定和蛋白质印迹分析评估阿瑞匹坦对NF-κB信号传导的抑制作用。
结果：NK-1R 的表达在 LPS 诱导的巨噬细胞中增加，表明该受体在炎症反应中的潜在作用。我们表明，阿瑞匹坦通过减少 ROS 的产生和 NOX-4 的表达来保护巨噬细胞免受氧化应激。此外，阿瑞匹坦通过介导 NF-κB 信号通路抑制促炎细胞因子和趋化因子的分泌。
结论： NK-1R受体拮抗剂阿瑞匹坦具有抗炎作用，表明阻断巨噬细胞NK-1R通路具有抑制炎症的作用。