In this study, we used ultra-performance liquid chromatography coupled with tandem mass spectrometry to assess the levels of eicosanoids from RAW264.7 macrophages treated with lipopolysaccharides (LPS) and 20(S)-ginsenoside Rg3 (Rg3). The production of nitric oxide (NO) and the secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were increased in inflammatory macrophages treated with LPS. Rg3 treatment, however, decreased the levels of NO, TNF-α, and IL-6 in activated macrophages. Eicosanoids, known as major metabolites correlated with inflammation, have pro- or anti-inflammatory activities. For a detailed characterization of the eicosanoids altered by treatment with LPS and Rg3, the eicosanoids were profiled by multiple reaction monitoring. A total of 69 macrophage eicosanoids were analyzed and the profiling dataset was statistically analyzed. Principal component and hierarchical cluster analyses differentiated control cells from cells treated with LPS, Rg3, or LPS + Rg3 for 12 or 24 h. Furthermore, 18 differentially regulated eicosanoids were found between macrophages treated with LPS for 24 h and those treated with LPS + Rg3 for 24 h (fold change > 2, p value < 0.05). These results indicate that Rg3 alters eicosanoid metabolism in activated macrophages treated with LPS. Furthermore, we also identified several eicosanoids correlated with the anti-inflammatory activity of Rg3.

在这项研究中，我们使用超高效液相色谱与串联质谱联用来评估脂多糖（LPS）和20（S）-人参皂苷Rg3（Rg3）处理的RAW264.7巨噬细胞中类花生酸的水平。用LPS处理的炎性巨噬细胞中一氧化氮（NO）的产生和肿瘤坏死因子-α（TNF-α）和白细胞介素6（IL-6）的分泌增加。然而，Rg3处理降低了活化巨噬细胞中NO，TNF-α和IL-6的水平。类花生酸，被称为与炎症相关的主要代谢产物，具有促炎或抗炎活性。为了详细描述通过LPS和Rg3处理改变的类花生酸的特征，通过多个反应监测对类花生酸进行了分析。总共分析了69个巨噬细胞类二十烷酸，并对分析数据集进行了统计分析。主成分和层次聚类分析可将分化后的对照细胞与经LPS，Rg3或LPS + Rg3处理12小时或24小时的细胞区分开。此外，在LPS处理24 h的巨噬细胞与LPS + Rg3处理24 h的巨噬细胞之间发现了18种差异调节的类花生酸（倍数> 2，p值<0.05）。这些结果表明，Rg3改变了LPS处理的活化巨噬细胞中类花生酸的代谢。此外，我们还鉴定了与Rg3的抗炎活性相关的几种类花生酸。