April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Oxygen Metabolism in Primary Human Lens Epithelial Cells; Association with Age, Diabetes and Glaucoma
Author Affiliations & Notes
  • Miyuki Kubota
    Ophthalmology & Visual Sciences, Washington Univ School of Medicine, St Louis, MO
    Ophthalmology, Keio University School of Medicine, Tokyo, Japan
  • Carla J Siegfried
    Ophthalmology & Visual Sciences, Washington Univ School of Medicine, St Louis, MO
  • Ying-Bo Shui
    Ophthalmology & Visual Sciences, Washington Univ School of Medicine, St Louis, MO
  • Fang Bai
    Ophthalmology & Visual Sciences, Washington Univ School of Medicine, St Louis, MO
  • Andrew J W Huang
    Ophthalmology & Visual Sciences, Washington Univ School of Medicine, St Louis, MO
  • David C Beebe
    Ophthalmology & Visual Sciences, Washington Univ School of Medicine, St Louis, MO
    Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO
  • Footnotes
    Commercial Relationships Miyuki Kubota, None; Carla Siegfried, None; Ying-Bo Shui, None; Fang Bai, None; Andrew Huang, None; David Beebe, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 6025. doi:
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      Miyuki Kubota, Carla J Siegfried, Ying-Bo Shui, Fang Bai, Andrew J W Huang, David C Beebe, ; Oxygen Metabolism in Primary Human Lens Epithelial Cells; Association with Age, Diabetes and Glaucoma. Invest. Ophthalmol. Vis. Sci. 2014;55(13):6025.

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

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Abstract

Purpose: The hypoxic environment around the lens is important for lens transparency. Lens epithelial cells (LECs) play a key role in lens metabolism. We measured oxygen consumption to assess the role of human LECs in maintaining hypoxia around the lens.

Methods: Baseline cellular respiration was measured in human primary LECs using a XF96 Analyzer (Seahorse Bioscience), which measures oxygen consumption rate (OCR) in 96-well plates. Following informed written consent, capsule epithelial cells were obtained from patients during cataract surgery and were divided into small explants in 96-well plates. Capsules were removed when LECs became confluent. OCR was normalized to the number of cells per well using rabbit LECs (NN1003A) as a standard. The effect on oxygen consumption of patient age, sex, race, and the presence of diabetes or glaucoma was assessed by using the Mann-Whitney U-test and multiple logistic regression analysis.

Results: Primary LECs were obtained from 65 patients of mean age 70 (± 10.8) years (44 women, 21 men). The OCR from donors aged 70 and over was lower than in those under 70 years (2.25 ± 0.03 vs. 2.96± 0.03 fMol/min/cell; ρ<0.05). Diabetic patients had lower OCR than non-diabetic patients (2.07 ± 0.05 vs. 2.83 ± 0.03 fMol/min/cell; ρ<0.05), and glaucoma patients had lower OCR than non-glaucoma patients (2.31 ± 0.04 vs. 2.94 ± 0.04fMol/min/cell; ρ<0.05). Further analysis using multiple logistic regression analysis confirmed that donors aged 70 and over (ρ<0.05), diabetic patients (ρ<0.01), and glaucoma patients (ρ<0.05) had significantly lower OCR, independent of other variables. Gender and race had no significant effect on OCR.

Conclusions: The lower O2 consumption rate of human LECs in older donors and patients with diabetes or glaucoma could contribute to cataract. Diabetes and glaucoma are especially important factors associated with decreased OCR, independent of age. Ongoing studies are examining pO2 at the anterior surface of the lens in vivo and oxygen consumption in the patient’s LECs.

Keywords: 467 clinical laboratory testing • 592 metabolism • 600 mitochondria  
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