July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Xanthohumol protects corneal epithelial cells against oxidative stress in vitro
Author Affiliations & Notes
  • Samatha Ankireddy
    Department of Molecular Pharmacology and Therapeutics, Loyola University Medical Center, Maywood, Illinois, United States
  • Harsh Nilesh Hariani
    Graduate Program in Neuroscience, Loyola University Medical Center, Maywood, Illinois, United States
  • Karoline Anna Orloff
    Department of Molecular Pharmacology and Therapeutics, Loyola University Medical Center, Maywood, Illinois, United States
  • Avinash Kolli
    Department of Ophthalmology, Loyola University Medical Center, Maywood, Illinois, United States
  • Jenni J Hakkarainen
    Research and Development, Experimentica Ltd., Kuopio, Finland
  • Anita K. Ghosh
    Graduate Program in Neuroscience, Loyola University Medical Center, Maywood, Illinois, United States
    Research and Development, eyeNOS Inc., Oak Park, Illinois, United States
  • Simon Kaja
    Department of Ophthalmology, Loyola University Medical Center, Maywood, Illinois, United States
    Research and Development, eyeNOS Inc., Oak Park, Illinois, United States
  • Footnotes
    Commercial Relationships   Samatha Ankireddy, None; Harsh Hariani, None; Karoline Orloff, None; Avinash Kolli, None; Jenni Hakkarainen, Experimentica Ltd. (F), Experimentica Ltd. (I), Experimentica Ltd. (E), Experimentica Ltd. (R), Experimentica Ltd. (S); Anita Ghosh, eyeNOS Inc. (F), eyeNOS Inc. (I), eyeNOS Inc. (P), eyeNOS Inc. (R), eyeNOS Inc. (S), K&P Scientific LLC (R); Simon Kaja, Experimentica Ltd. (F), Experimentica Ltd. (I), Experimentica Ltd. (R), Experimentica Ltd. (S), K&P Scientific LLC (F), K&P Scientific LLC (I), K&P Scientific LLC (P), K&P Scientific LLC (S)
  • Footnotes
    Support  Illinois Society for the Prevention of Blindness (AS, AKG), Department of Veterans Affairs (SK), Dr. John P. and Therese E. Mulcahy Endowed Professorship in Ophthalmology (SK) and the American Society for Pharmacology and Experimental Therapeutics Summer Undergraduate Research Fellowship (KAO). Richard A Perritt MD Charitable Foundation (SK)
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 5743. doi:
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    • Get Citation

      Samatha Ankireddy, Harsh Nilesh Hariani, Karoline Anna Orloff, Avinash Kolli, Jenni J Hakkarainen, Anita K. Ghosh, Simon Kaja; Xanthohumol protects corneal epithelial cells against oxidative stress in vitro. Invest. Ophthalmol. Vis. Sci. 2019;60(9):5743.

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

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Abstract

Purpose : Xanthohumol (Xn) is a natural compound found in the hops plant (Humulus Lupulus) and recognized as a potent antioxidant. The purpose of this study was to determine the cytoprotective effects of Xn against oxidative stress in human corneal epithelial cells in vitro.

Methods : Human Corneal Epithelial cells (HCE-T; Riken; Japan) were exposed to concentrations from 1 nM to 100 μM Xanthohumol and cell viability and proliferation were assessed using the lactate dehydrogenase (LDH) release and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) uptake assays in order to establish the in vitro toxicity profile. We next exposed HCE-T to chemically-induced exogenously-applied oxidative stress by treating cells with tert-butyl hydroperoxide (tBHP) for 6 hr. Cells were pretreated for 24 hr with Xanthohumol concentrations from 0.1 μM to 5 μM Xanthohumol, vehicle (0.01% DMSO) or remained untreated. Cytoprotective effects were assessed using MTT and LDH assays. Analysis of calcein-AM uptake and kinetics of the fluorescence were used to determine effects of Xn on P-glycoprotein 1, using cyclosporine A as a positive control.

Results : Xn did not show any cytotoxicity up to 10 μM as determined by LDH and MTT assays.
In cytoprotection experiments, pre-treatment with Xn resulted in a statistically significant dose-dependent protection of HCE-T cells from tBHP-induced oxidative stress compared with vehicle-treated cells. The EC50 of tBHP shifted from 15.2 ± 0.5 μM (untreated, n = 4) to 33.3 ± 3.4 μM (5 μM, n = 4, P < 0.01) in the MTT assay and, similarly, in the LDH assay (13.4 ± 0.4 μM vs.100.0 ± 11.7 μM; n = 4; P < 0.01). Changes in expression levels of components of the endogenous antioxidant system, including the intracellular target of Xn, Nrf2, in response to tBHP are currently being investigated.

Conclusions : Xn exerts potent dose-dependent cytoprotective effects against oxidative stress in HCE-T cells.
Our in vitro findings support testing Xn in preclinical models for DED, either as monotherapy or in combination with established anti-inflammatory treatment modalities.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

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