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Blockade of vascular adhesion protein-1 attenuates choroidal neovascularization.

Yoshikawa N, Noda K, Ozawa Y, Tsubota K, Mashima Y, Ishida S - Mol. Vis. (2012)

Bottom Line: VAP-1 inhibition significantly suppressed CNV formation in a dose-dependent manner and reduced macrophage infiltration into CNV lesions.Furthermore, VAP-1 blockade decreased the expression of ICAM-1 and MCP-1, both of which play a pivotal role in macrophage recruitment.VAP-1 inhibition may be a novel and potent therapeutic strategy in treating CNV formation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Retinal Cell Biology, Keio University School of Medicine, Tokyo, Japan.

ABSTRACT

Purpose: Vascular adhesion protein (VAP)-1 is an adhesion molecule elucidated as a mediator of the leukocyte recruitment cascade. The purpose of this study was to investigate the role of VAP-1 in ocular inflammatory neovascularization using a mouse laser-induced choroidal neovascularization (CNV) model.

Methods: CNV was induced with 532 nm laser irradiation in C57BL/6 mice, and production of VAP-1 protein in the retinal pigment epithelium (RPE) choroid during CNV formation was examined. CNV animals were treated with the specific VAP-1 inhibitor U-V002 or vehicle solution, and the volume of CNV tissue was evaluated with volumetric measurements. Macrophage infiltration into the CNV lesions was evaluated using two different techniques, flatmount staining and real-time polymerase chain reaction (PCR) for F4/80. The protein levels of intercellular adhesion molecule (ICAM)-1, monocyte chemoattractant protein (MCP)-1, P-selectin, and vascular endothelial growth factor (VEGF) in the RPE-choroid were measured with enzyme-linked immunosorbent assay (ELISA).

Results: VAP-1 inhibition significantly suppressed CNV formation in a dose-dependent manner and reduced macrophage infiltration into CNV lesions. Furthermore, VAP-1 blockade decreased the expression of ICAM-1 and MCP-1, both of which play a pivotal role in macrophage recruitment.

Conclusions: Our data suggest VAP-1 has an important role during ocular inflammatory neovascularization through leukocyte recruitment. VAP-1 inhibition may be a novel and potent therapeutic strategy in treating CNV formation.

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Localization and expression of VAP-1 in the choroid and CNV. A and B: Representative micrographs of a laser-induced CNV lesion. (A) Phase contrast image. (B) Fluorescent micrograph of VAP-1 (red) and cell nuclei (blue). Arrowheads and arrows indicate the localization of VAP-1 in the CNV and choroidal vessels, respectively. C: Immunoblotting analysis of VAP-1 and α-tubulin expression in the RPE-choroidal complex after laser injury.
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f1: Localization and expression of VAP-1 in the choroid and CNV. A and B: Representative micrographs of a laser-induced CNV lesion. (A) Phase contrast image. (B) Fluorescent micrograph of VAP-1 (red) and cell nuclei (blue). Arrowheads and arrows indicate the localization of VAP-1 in the CNV and choroidal vessels, respectively. C: Immunoblotting analysis of VAP-1 and α-tubulin expression in the RPE-choroidal complex after laser injury.

Mentions: To determine whether VAP-1 expression alters during CNV formation, we examined the localization of VAP-1 in CNV lesions with immunofluorescence staining and the time course of the VAP-1 protein levels with western blotting. VAP-1 was detected in endothelial cells of CNV and the choroidal vessels (Figure 1A). However, immunoblotting showed no change in the protein level of VAP-1 during CNV formation (Figure 1B).


Blockade of vascular adhesion protein-1 attenuates choroidal neovascularization.

Yoshikawa N, Noda K, Ozawa Y, Tsubota K, Mashima Y, Ishida S - Mol. Vis. (2012)

Localization and expression of VAP-1 in the choroid and CNV. A and B: Representative micrographs of a laser-induced CNV lesion. (A) Phase contrast image. (B) Fluorescent micrograph of VAP-1 (red) and cell nuclei (blue). Arrowheads and arrows indicate the localization of VAP-1 in the CNV and choroidal vessels, respectively. C: Immunoblotting analysis of VAP-1 and α-tubulin expression in the RPE-choroidal complex after laser injury.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3298450&req=5

f1: Localization and expression of VAP-1 in the choroid and CNV. A and B: Representative micrographs of a laser-induced CNV lesion. (A) Phase contrast image. (B) Fluorescent micrograph of VAP-1 (red) and cell nuclei (blue). Arrowheads and arrows indicate the localization of VAP-1 in the CNV and choroidal vessels, respectively. C: Immunoblotting analysis of VAP-1 and α-tubulin expression in the RPE-choroidal complex after laser injury.
Mentions: To determine whether VAP-1 expression alters during CNV formation, we examined the localization of VAP-1 in CNV lesions with immunofluorescence staining and the time course of the VAP-1 protein levels with western blotting. VAP-1 was detected in endothelial cells of CNV and the choroidal vessels (Figure 1A). However, immunoblotting showed no change in the protein level of VAP-1 during CNV formation (Figure 1B).

Bottom Line: VAP-1 inhibition significantly suppressed CNV formation in a dose-dependent manner and reduced macrophage infiltration into CNV lesions.Furthermore, VAP-1 blockade decreased the expression of ICAM-1 and MCP-1, both of which play a pivotal role in macrophage recruitment.VAP-1 inhibition may be a novel and potent therapeutic strategy in treating CNV formation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Retinal Cell Biology, Keio University School of Medicine, Tokyo, Japan.

ABSTRACT

Purpose: Vascular adhesion protein (VAP)-1 is an adhesion molecule elucidated as a mediator of the leukocyte recruitment cascade. The purpose of this study was to investigate the role of VAP-1 in ocular inflammatory neovascularization using a mouse laser-induced choroidal neovascularization (CNV) model.

Methods: CNV was induced with 532 nm laser irradiation in C57BL/6 mice, and production of VAP-1 protein in the retinal pigment epithelium (RPE) choroid during CNV formation was examined. CNV animals were treated with the specific VAP-1 inhibitor U-V002 or vehicle solution, and the volume of CNV tissue was evaluated with volumetric measurements. Macrophage infiltration into the CNV lesions was evaluated using two different techniques, flatmount staining and real-time polymerase chain reaction (PCR) for F4/80. The protein levels of intercellular adhesion molecule (ICAM)-1, monocyte chemoattractant protein (MCP)-1, P-selectin, and vascular endothelial growth factor (VEGF) in the RPE-choroid were measured with enzyme-linked immunosorbent assay (ELISA).

Results: VAP-1 inhibition significantly suppressed CNV formation in a dose-dependent manner and reduced macrophage infiltration into CNV lesions. Furthermore, VAP-1 blockade decreased the expression of ICAM-1 and MCP-1, both of which play a pivotal role in macrophage recruitment.

Conclusions: Our data suggest VAP-1 has an important role during ocular inflammatory neovascularization through leukocyte recruitment. VAP-1 inhibition may be a novel and potent therapeutic strategy in treating CNV formation.

Show MeSH
Related in: MedlinePlus