April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
DHA metabolism by human meibomian gland epithelial cells
Author Affiliations & Notes
  • Ulrike Hampel
    Medical faculty, Institute of Anatomy II, Erlangen, Germany
    University of New South Wales, School of Optometry and Vision Science, Sydney, NSW, Australia
  • Todd W Mitchell
    University of Wollongong, Illawara Health and Medical Research Institute and School of Health Sciences, Wollongong, NSW, Australia
  • Simon H Brown
    University of Wollongong, Illawara Health and Medical Research Institute and School of Health Sciences, Wollongong, NSW, Australia
  • Peta Snikeris
    University of Wollongong, Illawara Health and Medical Research Institute and School of Health Sciences, Wollongong, NSW, Australia
  • Garreis Fabian
    Medical faculty, Institute of Anatomy II, Erlangen, Germany
  • Friedrich P Paulsen
    Medical faculty, Institute of Anatomy II, Erlangen, Germany
  • Mark D P Willcox
    University of New South Wales, School of Optometry and Vision Science, Sydney, NSW, Australia
  • Footnotes
    Commercial Relationships Ulrike Hampel, Optima Pharmazeutische GmbH (F); Todd Mitchell, None; Simon Brown, None; Peta Snikeris, None; Garreis Fabian, None; Friedrich Paulsen, None; Mark Willcox, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1302. doi:https://doi.org/
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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)

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Abstract

Purpose: 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).

Methods: 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.

Results: 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.

Conclusions: 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.

Keywords: 583 lipids • 526 eyelid • 592 metabolism  
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