Add time:08/11/2019         Source:sciencedirect.com

This study examined the metabolism of dihomo-γ-linolenic acid (20:3(n−6)) in casein-elicited murine peritoneal macrophages. Cells were incubated with [14C]20:3(n−6) in the presence of 1% fetal bovine serum (FBS) or 0.025% bovine serum albumin (BSA), and the distribution and identity of membrane-bound and soluble products were determined. Approx. 70–80% of the [14C]20:3(n−6) was recovered in membrane phospholipids. The distribution of radiolabel in individual cellular phospholipids revealed a time-dependent (6 vs. 16 h) increase in the percentage of radiolabel esterified to phosphatidylethanolamine (PE). Analysis of cellular total lipids following transmethylation indicated that approx. 4, 2 and 9% of the incorporated 20:3(n−6), respectively, had been desaturated and elongated into 20:4(n−6), 22:4(n−6) and 22:3(n−6). When cells prelabeled for 16 h were incubated in the presence of the divalent cation ionophore, A23187, or zymosan for 30–60 min, two radiolabeled metabolites were isolated in the incubation supernatant. These metabolites were identified as 12-hydroxy-8,10,14- and 15-hydroxy-8,11,13-eicosatrienoic acids, as determined by reverse-phase and normal-phase high-performance liquid chromatography. The generation of monohydroxy fatty acids was notably absent in prelabeled quiescent cells and A23187-stimulated cells incubated with BW755C, a dual cyclooxygenase and lipoxygenase inhibitor. We conclude that casein-elicited murine peritoneal macrophages can extensively metabolize 20:3(n − 6) through Δ5desaturase, elongase and oxygenation reactions.

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