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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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Related in: MedlinePlus

Impact of VAP-1 blockade on inflammation-associated molecules. Bars indicate the average protein levels of (A) ICAM-1, (B) MCP-1, (C) P-selectin, and (D) VEGF in the RPE-choroidal complex obtained from laser-induced CNV animals treated with vehicle or VAP-1 inhibitor at 3 days after laser photocoagulation. Values are mean±SEM (n=9 to 10). *, p<0.05.
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f4: Impact of VAP-1 blockade on inflammation-associated molecules. Bars indicate the average protein levels of (A) ICAM-1, (B) MCP-1, (C) P-selectin, and (D) VEGF in the RPE-choroidal complex obtained from laser-induced CNV animals treated with vehicle or VAP-1 inhibitor at 3 days after laser photocoagulation. Values are mean±SEM (n=9 to 10). *, p<0.05.

Mentions: To further explore the mechanisms by which the VAP-1 blockade suppresses CNV formation, we measured the levels of the inflammation-associated molecules, ICAM-1, MCP-1, P-selectin, and VEGF in the RPE-choroid complex with or without VAP-1 inhibition (0.003%) at 3 days after laser treatment. The ICAM-1 (38.59±3.26 ng/mg, n=10) and MCP-1 (undetectable, n=10) protein levels in the RPE-choroidal complexes of mice treated with vehicle solution were significantly increased (ICAM-1, 134.05±9.28 pg/mg, n=10, p<0.05, Figure 4A; MCP-1, 28.41±3.97 pg/mg, n=9, p<0.05, Figure 4B) at 3 days after laser injury. The ICAM-1 (81.47±4.67 pg/mg) and MCP-1 (14.19±3.50 ng/mg) protein levels were significantly reduced in the RPE-choroidal complexes of the laser-treated animals that received the inhibitor compared with the vehicle controls (Figure 4A,B). In accord with our ELISA data, real-time PCR showed that the mRNA expression levels of Icam-1 and Mcp-1 were downregulated by 33.4% and 11.1%, respectively, in the animals treated with VAP-1 inhibitor (n=9) compared to that of the vehicle-treated animals (n=10).


Blockade of vascular adhesion protein-1 attenuates choroidal neovascularization.

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

Impact of VAP-1 blockade on inflammation-associated molecules. Bars indicate the average protein levels of (A) ICAM-1, (B) MCP-1, (C) P-selectin, and (D) VEGF in the RPE-choroidal complex obtained from laser-induced CNV animals treated with vehicle or VAP-1 inhibitor at 3 days after laser photocoagulation. Values are mean±SEM (n=9 to 10). *, p<0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Impact of VAP-1 blockade on inflammation-associated molecules. Bars indicate the average protein levels of (A) ICAM-1, (B) MCP-1, (C) P-selectin, and (D) VEGF in the RPE-choroidal complex obtained from laser-induced CNV animals treated with vehicle or VAP-1 inhibitor at 3 days after laser photocoagulation. Values are mean±SEM (n=9 to 10). *, p<0.05.
Mentions: To further explore the mechanisms by which the VAP-1 blockade suppresses CNV formation, we measured the levels of the inflammation-associated molecules, ICAM-1, MCP-1, P-selectin, and VEGF in the RPE-choroid complex with or without VAP-1 inhibition (0.003%) at 3 days after laser treatment. The ICAM-1 (38.59±3.26 ng/mg, n=10) and MCP-1 (undetectable, n=10) protein levels in the RPE-choroidal complexes of mice treated with vehicle solution were significantly increased (ICAM-1, 134.05±9.28 pg/mg, n=10, p<0.05, Figure 4A; MCP-1, 28.41±3.97 pg/mg, n=9, p<0.05, Figure 4B) at 3 days after laser injury. The ICAM-1 (81.47±4.67 pg/mg) and MCP-1 (14.19±3.50 ng/mg) protein levels were significantly reduced in the RPE-choroidal complexes of the laser-treated animals that received the inhibitor compared with the vehicle controls (Figure 4A,B). In accord with our ELISA data, real-time PCR showed that the mRNA expression levels of Icam-1 and Mcp-1 were downregulated by 33.4% and 11.1%, respectively, in the animals treated with VAP-1 inhibitor (n=9) compared to that of the vehicle-treated animals (n=10).

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