Fig. 5.

6BIGOE-loaded NPs downregulate prostanoid biosynthesis by inhibition of COX-2 expression in monocytes. A Schematic overview of LM biosynthetic pathways. B-D Human monocytes (10⁶/mL) were pretreated with vehicle (0.1% DMSO for 6BIGOE, or equivalent amount of blank NP dispersions), 6BIGOE or 6BIGOE-loaded NPs prepared by EDE, Cyrene, or PEG 400 method for 3 h before stimulation with 100 ng/mL LPS for 18 h. Formed LM were extracted from the supernatant by SPE and analyzed by UPLC-MS–MS. B Relative changes in the formation of single LMs from monocytes treated with 6BIGOE or 6BIGOE-loaded NPs at 0.3 µM versus vehicle-treated cells are shown in a heatmap, n = 5 separate donors, except 6BIGOE n = 4. Corresponding values ± SEM are included in Additional file 1: Figs. S5-S7. C Relative changes of LM groups in monocytes treated with 6BIGOE or 6BIGOE-loaded NPs versus vehicle-treated cells, n = 5 separate donors, except 6BIGOE n = 4. Statistical analysis was performed to compare 6BIGOE-treated cells with vehicle-treated cells (DMSO 0.1%) by one-way ANOVA with Dunnett’s correction on log-transformed data, and unpaired, two-sided Student’s t-test to compare 6BIGOE-loaded NPs with respective blank NP controls. D PGE₂ formation in 6BIGOE- or 6BIGOE-loaded NPs-treated monocytes vs. vehicle-treated monocytes; n = 5 separate donors, except 6BIGOE n = 4. Statistical analysis was performed on log-transformed data using one-way ANOVA with Dunnett’s correction for multiple comparisons of 6BIGOE with control (DMSO 0.1%). Unpaired, two-tailed Student’s t-test was applied for comparison of 6BIGOE-NPs with respective NP blank controls. E COX-2 protein expression was assessed by Western Blot and densitometric analysis thereof, normalized to β-actin. Results are individual values represented in a box and whisker plot, n = 4 separate donors. Statistical analysis displayed was performed using one-way ANOVA for multiple comparisons with Holm-Šídák post-hoc test