PGC-1α Induces Human RPE Oxidative Metabolism and Antioxidant Capacity
| Autor: | Magali Saint-Geniez, Zoltan Arany, Arogya Khadka, Daniel Diaz-Aguilar, Glenn C. Rowe, Jared Iacovelli, Carrie Spencer |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Cellular respiration Blotting Western retinal pigment epithelium Oxidative phosphorylation Mitochondrion Biology medicine.disease_cause Polymerase Chain Reaction Oxidative Phosphorylation Cell Line 03 medical and health sciences PGC-1 Macular Degeneration Mediator Coactivator medicine Humans age-related macular degeneration Heat-Shock Proteins chemistry.chemical_classification Regulation of gene expression Reactive oxygen species Cell Death Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha eye diseases Cell biology Mitochondria Oxidative Stress 030104 developmental biology chemistry Retinal Cell Biology Gene Expression Regulation RNA sense organs Reactive Oxygen Species metabolism Oxidative stress Transcription Factors |
| Zdroj: | Investigative Ophthalmology & Visual Science |
| ISSN: | 1552-5783 0146-0404 |
| Popis: | Purpose Oxidative stress and metabolic dysregulation of the RPE have been implicated in AMD; however, the molecular regulation of RPE metabolism remains unclear. The transcriptional coactivator, peroxisome proliferator-activated receptor-gamma coactivator 1α (PGC-1α) is a powerful mediator of mitochondrial function. This study examines the ability of PGC-1α to regulate RPE metabolic program and oxidative stress response. Methods Primary human fetal RPE (hfRPE) and ARPE-19 were matured in vitro using standard culture conditions. Mitochondrial mass of RPE was measured using MitoTracker staining and citrate synthase activity. Expression of PGC-1 isoforms, RPE-specific genes, oxidative metabolism proteins, and antioxidant enzymes was analyzed by quantitative PCR and Western blot. Mitochondrial respiration and fatty-acid oxidation were monitored using the Seahorse extracellular flux analyzer. Expression of PGC-1α was increased using adenoviral delivery. ARPE-19 were exposed to hydrogen peroxide to induce oxidative stress. Reactive oxygen species were measured by CM-H2DCFDA fluorescence. Cell death was analyzed by LDH release. Results Maturation of ARPE-19 and hfRPE was associated with significant increase in mitochondrial mass, expression of oxidative phosphorylation (OXPHOS) genes, and PGC-1α gene expression. Overexpression of PGC-1α increased expression of OXPHOS and fatty-acid β-oxidation genes, ultimately leading to the potent induction of mitochondrial respiration and fatty-acid oxidation. PGC-1α gain of function also strongly induced numerous antioxidant genes and, importantly, protected RPE from oxidant-mediated cell death without altering RPE functions. Conclusions This study provides important insights into the metabolic changes associated with RPE functional maturation and identifies PGC-1α as a potent driver of RPE mitochondrial function and antioxidant capacity. |
| Databáze: | OpenAIRE |
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