Abstract | Purpose: Methods: Male Wistar rats (n = 7, unless otherwise specified for specific experiments) were administered an oral dose of 30 mg of Nω-nitro- L-arginine methyl ester ( L-NAME) per kilogram of bodyweight and day for 3 weeks; chronic administration with L-NAME is a validated experimental approach resulting in severe hypertension secondary to nitric oxide (NO) depletion and subsequent vasoconstriction in the systemic circulation. Upon treatment completion, histomorphometric studies, NADPH oxidase activity, and ROS production were measured in eyecup homogenates and paraffin-embedded sections from control and L-NAME-treated animals. In addition, immunohistofluorescence, western blotting, and real-time PCR (RT-qPCR) analyses were performed in the eye and the retina to evaluate the expression of i) NADPH oxidase main isoforms (NOX1, NOX2, and NOX4) and subunits (p22phox and p47phox); ii) glial fibrillary acidic protein (GFAP), as a marker of microglial activation in the retina; iii) antioxidant enzymes; and iv) endothelial constitutive (eNOS) and inflammation inducible (iNOS) nitric oxide synthase isoforms, and nitrotyrosine as a versatile biomarker of oxidative stress. Results: Conclusions: The present study demonstrated structural changes in the retinas of the L-NAME-treated hypertensive animals and strengthens the importance of NADPH oxidase as a major ROS-generating enzyme system in the oxidative and inflammatory processes surrounding hypertensive eye diseases. These observations might contribute to unveiling pathogenic mechanisms responsible for developing ocular disturbances in the context of severe hypertension.
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Authors | Álvaro Santana-Garrido, Claudia Reyes-Goya, Carmen Fernández-Bobadilla, Antonio J Blanca, Helder André, Alfonso Mate, Carmen M Vázquez |
Journal | Molecular vision
(Mol Vis)
Vol. 27
Pg. 161-178
( 2021)
ISSN: 1090-0535 [Electronic] United States |
PMID | 33907371
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2021 Molecular Vision. |
Chemical References |
- Biomarkers
- Enzyme Inhibitors
- GFAP protein, rat
- Glial Fibrillary Acidic Protein
- RNA, Messenger
- Reactive Oxygen Species
- Superoxides
- Nitric Oxide
- Nitric Oxide Synthase Type II
- Nitric Oxide Synthase Type III
- Nos2 protein, rat
- Nos3 protein, rat
- NADPH Oxidases
- NG-Nitroarginine Methyl Ester
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Topics |
- Animals
- Biomarkers
(metabolism)
- Blood Pressure
(drug effects)
- Blotting, Western
- Enzyme Inhibitors
(toxicity)
- Glial Fibrillary Acidic Protein
(metabolism)
- Male
- NADPH Oxidases
(genetics, metabolism)
- NG-Nitroarginine Methyl Ester
(toxicity)
- Nitric Oxide
(metabolism)
- Nitric Oxide Synthase Type II
(metabolism)
- Nitric Oxide Synthase Type III
(metabolism)
- Ocular Hypertension
(chemically induced, enzymology)
- Oxidative Stress
(physiology)
- RNA, Messenger
(genetics)
- Rats
- Rats, Wistar
- Reactive Oxygen Species
(metabolism)
- Real-Time Polymerase Chain Reaction
- Retina
(drug effects, pathology)
- Superoxides
(metabolism)
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