Mitochondrial Oxygen Metabolism in Primary Human Lens Epithelial Cells: Association with Age, Diabetes and Glaucoma

Publication date: Available online 19 July 2016
Source:Free Radical Biology and Medicine
Author(s): M. Kubota, Y.B. Shui, M. Liu, F. Bai, A.J. Huang, N. Ma, D.C. Beebe, C.J. Siegfried
PurposeThe hypoxic environment around the lens is important for maintaining 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, as well as the impact of systemic and ocular diagnosis on these cells.MethodsBaseline cellular respiration was measured in rabbit LECs (NN1003A), canine kidney epithelial cells (MDCK), trabecular meshwork cells (TM-5), and bovine corneal endothelial cells (CCEE) using a XF96 Extracellular Flux Analyzer (Seahorse Bioscience, North Billerica, MA), which measures oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) in vitro. Following informed written consent, lens 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 as a standard. The effect of patient age, sex, race, and presence of diabetes or glaucoma on oxygen consumption was assessed by using the Mann-Whitney U test and multivariate regression analysis.ResultsPrimary LECs were obtained from 69 patients. The OCR from donors aged 70 and over was lower than that of those under 70 years (2.21 ± 1.037 vs. 2.86 ± 1.383 fmol/min/cell; p<0.05). Diabetic patients had lower OCR than non-diabetic patients (2.02 ± 0.911 vs. 2.79 ± 1.332 fmol/min/cell; p<0.05), and glaucoma patients had lower OCR than non-glaucoma patients (2.27 ± 1.19 vs. 2.83 ± 1.286 fmol/min/cell; p<0.05). Multivariate regression analysis confirmed that donors aged 70 and over (p<0.05), diabetic patients (p<0.01), and glaucoma patients (p<0.05) had significantly lower OCR, independent of other variables. Gender and race had no significant effect on OCR.ConclusionsThe lower oxygen consumption rate of human LECs in older donors and patients with diabetes or glaucoma could contribute to cataract development. Diabetes and glaucoma are particularly 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.The hypoxic environment around the lens is important for maintaining lens transparency. Lens epithelial cells (LECs) play a key role in lens metabolism. Using a XF96 Extracellular Flux Analyzer (Seahorse Bioscience), we measured oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of various cell types (rabbit LECs, canine kidney epithelial cells, trabecular meshwork cells, bovine corneal endothelial cells, and human LECs from 69 patients of cataract surgery) to assess the role of human LECs in maintaining hypoxia around the lens, as well as the impact of systemic and ocular diagnosis on human LECs. Multivariate regression analysis confirmed that donors aged 70 and over (p<0.05), diabetic patients (p<0.01), and glaucoma patients (p<0.05) had significantly lower OCR, independent of other variables. Gender and race had no significant effect on OCR. The lower OCR of human LECs in older donors and patients with diabetes or glaucoma could contribute to cataract development.



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