June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
Prostaglandins alter expression of select long-chain cholesteryl esters produced by human meibomian gland epithelial cells in culture
Author Affiliations & Notes
  • Jillian F. Ziemanski
    The School of Optometry, The University of Alabama at Birmingham, Hoover, Alabama, United States
  • Landon Wilson
    Pharmacology & Toxicology, University of Alabama at Birmingham, Alabama, United States
  • Stephen Barnes
    Pharmacology & Toxicology, University of Alabama at Birmingham, Alabama, United States
  • Kelly K Nichols
    The School of Optometry, The University of Alabama at Birmingham, Hoover, Alabama, United States
  • Footnotes
    Commercial Relationships   Jillian Ziemanski, None; Landon Wilson, None; Stephen Barnes, None; Kelly Nichols, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 129. doi:
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      Jillian F. Ziemanski, Landon Wilson, Stephen Barnes, Kelly K Nichols; Prostaglandins alter expression of select long-chain cholesteryl esters produced by human meibomian gland epithelial cells in culture. Invest. Ophthalmol. Vis. Sci. 2020;61(7):129.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Ocular surface disease is a common comorbidity in glaucoma patients treated with prostaglandin analogs. To date, however, there is little understanding of the physiologic roles that prostaglandins have on the meibomian glands. Therefore, human meibomian gland epithelial cells (HMGECs) were treated with two common prostaglandins (PGE2 and PGF2α) to evaluate their effects on cell viability and lipid expression.

Methods : Differentiated HMGECs were stained with antibodies against PGE2 (EP1, EP2, EP3, EP4) and PGF2α (FP) receptors and imaged with the Zeiss Axioplan-2 Microscope (Jena, Germany). For cell viability and lipidomic analyses, differentiated HMGECs were cultured with PGE2 or PGF (1 to 1000 nM) for three hours. Following culture, cell viability was assessed with the Cell Titer Glo Assay (Promega, Madison, WI). Lipid extracts were concentrated two-fold and directly infused into a Triple TOF 5600 mass spectrometer (SCIEX, Framingham, MA) with electrospray ionization in the positive ion mode. Meibum-relevant lipids were defined to be wax esters (WE) with carbon number (nc) ≥ 32 and cholesteryl esters (CE) with nc ≥ 16. Lipids are labeled as nc: # double bonds.

Results : Differentiated HMGECs express three receptors for PGE2 (EP1, EP2, and EP4) and the only receptor for PGF (FP). None of the concentrations of PGE2 (p > 0.73 for all) or PGF (p > 0.99 for all) affected cell viability. When evaluating the lipidome, there were 26 CEs and 0 WEs that met the criteria for inclusion. Increasing concentrations of PGE2 were associated with a dose-dependent decrease in CE 22:1 (mean fold change [FC] = 0.89 per ten-fold change in dose, p = 0.03). Both the 1000 and 100 nM concentrations of PGE2 significantly reduced CE 24:0 relative to control (mean FC = 0.47, p < 0.03 for both). For PGF2α, the 10 nM concentration upregulated CE 16:0 relative to control (FC = 1.31, p = 0.01). All other CEs remained unchanged across all concentrations of prostaglandins.

Conclusions : Receptors for both PGE2 and PGF are expressed on HMGECs. Neither PGE2 or PGF affects cell viability; however, both molecules are capable of altering the expression of select cholesteryl esters. It is possible that prostaglandin analogs elicit molecular changes to human meibum and the tear film, which may represent one possible explanation for the concurrence of glaucoma and ocular surface disease.

This is a 2020 ARVO Annual Meeting abstract.

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