Fig. 2: Inhibition of CPT1 increases secretion of 12/15-LOX-derived eicosanoids from RAW cells overexpressing Alox15, while RAW cells rapidly consume exogenous 12-HETE, secreting two tetranor metabolites that are themselves metabolized by mitochondrial β-oxidation.
From: Oxylipin metabolism is controlled by mitochondrial β-oxidation during bacterial inflammation
a Blockade of CPT1 leads to a significant elevation of 12/15-LOX-derived oxylipins in RAW cells. RAWmock and RAWAlox15 cells were cultured with etomoxir (25 μM) and/or LPS (100 ng/ml) for 24 h before harvest and SPE extraction of supernatant for oxylipin analysis using LC/MS/MS. Bar charts showing how oxylipins from 12/15-LOX are impacted by etomoxir (n = 6, separate wells of cells, mean ± SEM). b CPT1 inhibition prevents the metabolism of 12(S) or 12(R)HETE and their tetranor triene and diene metabolites. RAW cells were supplemented with 1.5 μg 12(S) or 12(R)-HETE/106 cells for 3 h with/without etomoxir (25 μM), then supernatants were analyzed for levels of 12-HETE and its triene and diene tetranor products using LC/MS/MS (n = 3 (LPS alone) or 4 (all other samples), mean ± SEM, separate wells of cells). For all panels, comparisons are with/without etomoxir, one-way ANOVA with Tukey post hoc test, stats are shown for the effect of etomoxir only, where significant.
