Corneal oxidative damage in keratoconus cells due to decreased oxidant elimination from modified expression levels of SOD enzymes, PRDX6, SCARA3, CPSF3, and FOXM1.
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 fibro...
| Publicado en: | Journal of Ophthalmic & Vision Research Vol. 14; no. 1; pp. 62 - 71 |
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| Autores principales: | , , , , , , |
| Formato: | pictorial research tables/charts Journal Article |
| Publicado: |
Knowledge E DMCC
Jan-Mar2019
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=134601118&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 134601118 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 20082010 AY5S jtl: Journal of Ophthalmic & Vision Research issn: 20082010 maglogo: N pubinfo: dt: Jan-Mar2019 vid: 14 iid: 1 pid: 35682 pub: Knowledge E DMCC artinfo: ui: 134601118 134601118 134601118 10.4103/jovr.jovr_80_18 134601118 ppf: 62 ppct: 9 formats: fmt: – @attributes: type: T – @attributes: type: P tig: atl: Corneal oxidative damage in keratoconus cells due to decreased oxidant elimination from modified expression levels of SOD enzymes, PRDX6, SCARA3, CPSF3, and FOXM1. aug: au: Atilano, Shari Lee, Daniel Fukuhara, Paula Chwa, Marilyn Nesburn, Anthony Udar, Nitin Kenney, Cristina affil: Department of Ophthalmology, Gavin Herbert Eye Institute, University of California Irvine, Irvine, CA sug: subj: Corneal Injuries Pathology Keratoconus Pathology Oxidative Stress Oxidants Adverse Effects Gene Expression Superoxide Dismutase Metabolism Human Comparative Studies In Vitro Studies Reactive Oxygen Species Reactive Nitrogen Species Polymerase Chain Reaction Methods Biological Assay Methods Immunohistochemistry Descriptive Statistics ab: 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 H2DCFDA 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. pubtype: Academic Journal doctype: pictorial research tables/charts Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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