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Ulrike Hampel, Todd W Mitchell, Simon H Brown, Peta Snikeris, Garreis Fabian, Friedrich P Paulsen, Mark D P Willcox; DHA metabolism by human meibomian gland epithelial cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1302. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the metabolism of the omega 3 fatty acid docosahexaenoic acid (DHA) by meibomian gland epithelial cells (MG cells) and production of one of its metabolites Resolvin D1 (RvD1).
MG cells were differentiated in DMEM containing EGF and DHA for up to 3 days. Optimal DHA concentration was assessed by measuring lipid droplet accumulation with Sudan III staining. Gene expression of cyclooxygenase-2 (COX-2) and 15-lipoxygenase (15-LOX) after DHA incubation was analyzed by real-time PCR. RvD1 concentrations of MG cell extracts were measured by enzyme immune assay. Lipid cell extracts were analyzed by automated, chip-based nanoelectrospray ionization tandem mass spectrometry.
Lipid droplet accumulation was increased by 100 µM DHA supplementation. COX-2 mRNA expression decreased to 31% compared to control during DHA stimulation after 3 days (p < 0.05). 15-LOX mRNA expression was reduced by 28% after three days of DHA incubation (p < 0.05). Both mRNA levels were not significantly changed after 1 day incubation with DHA. The concentration of RvD1 was elevated 2-fold after DHA incubation (20.7 vs. 10.4 pg/mg total protein; p < 0.05). Total triglyceride (TAG) concentration was elevated during DHA supplementation.
DHA is esterified to TAG by MG cells. Furthermore, DHA supplementation supports anti-inflammatory effects by decreasing COX-2 expression and increasing the concentration of RvD1.
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