The effect of triamcinolone acetonide on laser-induced choroidal neovascularization in mice using a hypoxia visualization bio-imaging probe.

Abstract

Hypoxic stress is a risk factor of ocular neovascularization. Hypoxia visualization may provide clues regarding the underlying cause of angiogenesis. Recently, we developed a hypoxia-specific probe, protein transduction domain-oxygen-dependent degradation domain-HaloTag-Rhodamine (POH-Rhodamine). In this study, we observed the localization of HIF-1α proteins by immunohistochemistry and the fluorescence of POH-Rhodamine on RPE-choroid flat mounts. Moreover, we compared the localization of POH-Rhodamine with pimonidazole which is a standard reagent for detecting hypoxia. Next, we investigated the effects of triamcinolone acetonide (TAAC) against visual function that was evaluated by recording electroretinogram (ERG) and choroidal neovascularization (CNV) development. Mice were given laser-induced CNV using a diode laser and treated with intravitreal injection of TAAC. Finally, we investigated POH-Rhodamine on CNV treated with TAAC. In this study, the fluorescence of POH-Rhodamine and HIF-1α were co-localized in laser-irradiated sites, and both the POH-Rhodamine and pimonidazole fluorescent areas were almost the same. Intravitreal injection of TAAC restored the reduced ERG b-wave but not the a-wave and decreased the mean CNV area. Furthermore, the area of the POH-Rhodamine-positive cells decreased. These findings indicate that POH-Rhodamine is useful for evaluating tissue hypoxia in a laser-induced CNV model, suggesting that TAAC suppressed CNV through tissue hypoxia improvement.

• Authors: Shinsuke Takata, Tomomi Masuda, Shinsuke Nakamura, Takahiro Kuchimaru, Kazuhiro Tsuruma, Masamitsu Shimazawa, Hideko Nagasawa, Shinae Kizaka-Kondoh, Hideaki Hara
• Journal: Scientific reports (Sci Rep) Vol. 5 Pg. 9898 (Apr 30 2015) ISSN: 2045-2322 [Electronic] England
• PMID: 25927172 (Publication Type: Journal Article)

Chemical References

• Hypoxia-Inducible Factor 1, alpha Subunit
• Molecular Probes
• Rhodamines
• Triamcinolone Acetonide
• Oxygen

Topics

• Animals
• Choroidal Neovascularization (drug therapy, metabolism, pathology)
• Disease Models, Animal
• Electroretinography (methods)
• Hypoxia (metabolism, pathology)
• Hypoxia-Inducible Factor 1, alpha Subunit (metabolism)
• Lasers (adverse effects)
• Male
• Mice
• Mice, Inbred C57BL
• Molecular Probes (metabolism)
• Oxygen (metabolism)
• Protein Structure, Tertiary (drug effects)
• Proteolysis (drug effects)
• Rhodamines (metabolism)
• Triamcinolone Acetonide (pharmacology)