Characterization of the changes in eicosanoid profiles of activated macrophages treated with 20(S)-ginsenoside Rg3

- UPLC-MS/MS was used to assess the levels of various eicosanoids in the activated macrophages treated with Rg3.
- A total of 69 eicosanoids were analyzed in activated macrophages treated with LPS.
- We identified 18 eicosanoids correlated with the anti-inflammatory activity of Rg3.
- Rg3-dependent regulation of eicosanoids are important to elucidate the anti-inflammatory mechanisms of Rg3.

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.