Corneal Oxidative Damage in Keratoconus Cells due to Decreased Oxidant Elimination from Modified Expression Levels of SOD Enzymes, PRDX6, SCARA3, CPSF3, and FOXM1

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Atilano Shari R.; Lee Daniel H.; Fukuhara Paula S.; Chwa Marilyn; Nesburn Anthony B.; Udar Nitin; Kenney M. Cristina

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Journal Paper

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Journal: Journal of Ophthalmic and Vision Research Year:2019 | Volume:14 | Issue:1 Page(s): 62-70

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Title

Corneal Oxidative Damage in Keratoconus Cells due to Decreased Oxidant Elimination from Modified Expression Levels of SOD Enzymes, PRDX6, SCARA3, CPSF3, and FOXM1

Author(s)

Keywords

CPSF3

FOXM1

Keratoconus

PRDX6

SCARA3

SOD

Abstract

Purpose: To compare the levels of gene expression for enzymes involved in production and elimination of reactive oxygen/nitrogen species (ROS/RNS) in normal human corneal cells (NL cells) with those in human corneal cells with Keratoconus (KC cells) in vitro. Methods: Primary NL and KC stromal fibroblast cultures were incubated with apocynin (an inhibitor of NADPH oxidase) or N‑ nitro‑ L‑ arginine (N‑ LLA; an inhibitor of nitric oxide synthase). ROS/RNS levels were measured using an H2 DCFDA fluorescent assay. The RT2 Profiler™ PCR Array for Oxidative Stress and Antioxidant Defense was used for initial screening of the NL and KC cultures. Transcription levels for genes related to production or elimination of ROS/RNS were analyzed using quantitative PCR. Immunohistochemistry was performed on 10 intact human corneas using antibodies against SCARA3 and CPSF3. Results: Array screening of 84 antioxidant-related genes identified 12 genes that were differentially expressed between NL and KC cultures. Compared with NL cells, quantitative PCR showed that KC cells had decreased expression of antioxidant genes SCARA3 isoform 2 (0. 59‑ fold, P = 0. 02) and FOXM1 isoform 1 (0. 61‑ fold, P = 0. 03). KC cells also had downregulation of the antioxidant genes SOD1 (0. 4‑ fold, P = 0. 0001) and SOD3 (0. 37‑ fold, P = 0. 02) but increased expression of SOD2 (3. 3‑ fold, P < 0. 0001), PRDX6 (1. 47‑ fold, P = 0. 01), and CPSF3 (1. 44‑ fold, P = 0. 02). Conclusion: The difference in expression of antioxidant enzymes between KC and NL suggests that the oxidative stress imbalances found in KC are caused by defects in ROS/RNS removal rather than increased ROS/RNS production.

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APA: Copy

Atilano, Shari R., Lee, Daniel H., Fukuhara, Paula S., Chwa, Marilyn, Nesburn, Anthony B., Udar, Nitin, & Kenney, M. Cristina. (2019). Corneal Oxidative Damage in Keratoconus Cells due to Decreased Oxidant Elimination from Modified Expression Levels of SOD Enzymes, PRDX6, SCARA3, CPSF3, and FOXM1. JOURNAL OF OPHTHALMIC AND VISION RESEARCH, 14(1), 62-70. SID. https://sid.ir/paper/311743/en

Vancouver: Copy

Atilano Shari R., Lee Daniel H., Fukuhara Paula S., Chwa Marilyn, Nesburn Anthony B., Udar Nitin, Kenney M. Cristina. Corneal Oxidative Damage in Keratoconus Cells due to Decreased Oxidant Elimination from Modified Expression Levels of SOD Enzymes, PRDX6, SCARA3, CPSF3, and FOXM1. JOURNAL OF OPHTHALMIC AND VISION RESEARCH[Internet]. 2019;14(1):62-70. Available from: https://sid.ir/paper/311743/en

IEEE: Copy

Shari R. Atilano, Daniel H. Lee, Paula S. Fukuhara, Marilyn Chwa, Anthony B. Nesburn, Nitin Udar, and M. Cristina Kenney, “Corneal Oxidative Damage in Keratoconus Cells due to Decreased Oxidant Elimination from Modified Expression Levels of SOD Enzymes, PRDX6, SCARA3, CPSF3, and FOXM1,” JOURNAL OF OPHTHALMIC AND VISION RESEARCH, vol. 14, no. 1, pp. 62–70, 2019, [Online]. Available: https://sid.ir/paper/311743/en

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