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Decursin inhibits retinal neovascularization via suppression of VEGFR-2 activation.

Kim JH, Kim JH, Lee YM, Ahn EM, Kim KW, Yu YS - Mol. Vis. (2009)

Bottom Line: We evaluated the inhibitory effect of decursin on retinal neovascularization.Decursin significantly inhibited VEGF-induced proliferation, migration, and the formation of capillary-like networks of retinal endothelial cells in a dose-dependent manner.Moreover, the upregulation of glial fibrillary acidic protein expression was not detected in Mueller cells.

View Article: PubMed Central - PubMed

Affiliation: Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Department of Ophthalmology, College of Medicine, Seoul National University and Seoul Artificial Eye Center Clinical Research Institute, Seoul National University Hospital, Seoul 151-744, Korea.

ABSTRACT

Purpose: Pathologic angiogenesis in the retina leads to the catastrophic loss of vision. Retinopathy of prematurity (ROP), a vasoproliferative retinopathy, is a leading cause of blindness in children. We evaluated the inhibitory effect of decursin on retinal neovascularization.

Methods: Anti-angiogenic activity of decursin was evaluated by vascular endothelial growth factor (VEGF)-induced proliferation, migration, and in vitro tube formation assay of human retinal microvascular endothelial cells (HRMECs). We also used western blot analysis to assess inhibition of vascular endothelial growth factor receptor-2 (VEGFR-2) phosphorylation by decursin. After intravitreal injection of decursin in a mouse model of ROP, retinal neovascularization was examined by fluorescence angiography and vessel counting in cross-sections. The toxicity of decursin was evaluated through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in HRMECs as well as histologic and immunohistochemistry examination for glial fibrillary acidic protein in the retina.

Results: Decursin significantly inhibited VEGF-induced proliferation, migration, and the formation of capillary-like networks of retinal endothelial cells in a dose-dependent manner. Decursin inhibited VEGF-induced phosphorylation of VEGFR-2, blocking the VEGFR-2 signaling pathway. When intravitreously injected, decursin dramatically suppressed retinal neovascularization in a mouse model of ROP. Even in a high concentration, decursin never induced any structural or inflammatory changes to cells in retinal or vitreous layers. Moreover, the upregulation of glial fibrillary acidic protein expression was not detected in Mueller cells.

Conclusions: Our data suggest that decursin may be a potent anti-angiogenic agent targeting the VEGFR-2 signaling pathway, which significantly inhibits retinal neovascularization without retinal toxicity and may be applicable in various other vasoproliferative retinopathies as well.

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Decursin inhibits retinal neovascularization in oxygen-induced retinopathy. A, B: Retinal vasculature in control mice and decursin-treated mice with oxygen-induced retinopathy (OIR) was evaluated by fluorescence angiography using fluorescein-conjugated dextran. Whole-mount retinal preparation from postnatal day 17 (P17) control mice (A) and mice subjected to OIR and treated with 10 µM decursin (B) was performed after 1 h of perfusion with fluorescein-conjugated dextran. Arrows indicate neovascular tufts of intravitreous neovascularization. Figures were selected as representative data from three independent experiments with similar results. Scale bars equal 50 µm. Hematoxylin-stained cross-sections were prepared from P17 control mice (C) and mice subjected to OIR and treated with 10 μM of decursin (D). Arrows indicate the vascular lumens of new vessels growing into the vitreous. Figures were selected as representative data from three independent experiments with similar results. Scale bars equal 100 µm. E: Each value represents the mean (±SD) of three independent experiments. The asterisk indicates a p<0.05.
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f3: Decursin inhibits retinal neovascularization in oxygen-induced retinopathy. A, B: Retinal vasculature in control mice and decursin-treated mice with oxygen-induced retinopathy (OIR) was evaluated by fluorescence angiography using fluorescein-conjugated dextran. Whole-mount retinal preparation from postnatal day 17 (P17) control mice (A) and mice subjected to OIR and treated with 10 µM decursin (B) was performed after 1 h of perfusion with fluorescein-conjugated dextran. Arrows indicate neovascular tufts of intravitreous neovascularization. Figures were selected as representative data from three independent experiments with similar results. Scale bars equal 50 µm. Hematoxylin-stained cross-sections were prepared from P17 control mice (C) and mice subjected to OIR and treated with 10 μM of decursin (D). Arrows indicate the vascular lumens of new vessels growing into the vitreous. Figures were selected as representative data from three independent experiments with similar results. Scale bars equal 100 µm. E: Each value represents the mean (±SD) of three independent experiments. The asterisk indicates a p<0.05.

Mentions: To investigate whether decursin could reduce retinal neovascularization, we injected 10 µM decursin intravitreously on P14 in OIR, and qualitatively and quantitatively analyzed the anti-angiogenic activity of decursin on retinal neovascularization. First, to investigate the anti-angiogenic activity of decursin on retinal neovascularization in OIR, we performed fluorescence angiography. On P17 in OIR, many neovascular tufts at the border of vascularized and nonvascularized retina were easily detected (Figure 3A), whereas they were significantly reduced in decursin-treated mice (Figure 3B).


Decursin inhibits retinal neovascularization via suppression of VEGFR-2 activation.

Kim JH, Kim JH, Lee YM, Ahn EM, Kim KW, Yu YS - Mol. Vis. (2009)

Decursin inhibits retinal neovascularization in oxygen-induced retinopathy. A, B: Retinal vasculature in control mice and decursin-treated mice with oxygen-induced retinopathy (OIR) was evaluated by fluorescence angiography using fluorescein-conjugated dextran. Whole-mount retinal preparation from postnatal day 17 (P17) control mice (A) and mice subjected to OIR and treated with 10 µM decursin (B) was performed after 1 h of perfusion with fluorescein-conjugated dextran. Arrows indicate neovascular tufts of intravitreous neovascularization. Figures were selected as representative data from three independent experiments with similar results. Scale bars equal 50 µm. Hematoxylin-stained cross-sections were prepared from P17 control mice (C) and mice subjected to OIR and treated with 10 μM of decursin (D). Arrows indicate the vascular lumens of new vessels growing into the vitreous. Figures were selected as representative data from three independent experiments with similar results. Scale bars equal 100 µm. E: Each value represents the mean (±SD) of three independent experiments. The asterisk indicates a p<0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC2743803&req=5

f3: Decursin inhibits retinal neovascularization in oxygen-induced retinopathy. A, B: Retinal vasculature in control mice and decursin-treated mice with oxygen-induced retinopathy (OIR) was evaluated by fluorescence angiography using fluorescein-conjugated dextran. Whole-mount retinal preparation from postnatal day 17 (P17) control mice (A) and mice subjected to OIR and treated with 10 µM decursin (B) was performed after 1 h of perfusion with fluorescein-conjugated dextran. Arrows indicate neovascular tufts of intravitreous neovascularization. Figures were selected as representative data from three independent experiments with similar results. Scale bars equal 50 µm. Hematoxylin-stained cross-sections were prepared from P17 control mice (C) and mice subjected to OIR and treated with 10 μM of decursin (D). Arrows indicate the vascular lumens of new vessels growing into the vitreous. Figures were selected as representative data from three independent experiments with similar results. Scale bars equal 100 µm. E: Each value represents the mean (±SD) of three independent experiments. The asterisk indicates a p<0.05.
Mentions: To investigate whether decursin could reduce retinal neovascularization, we injected 10 µM decursin intravitreously on P14 in OIR, and qualitatively and quantitatively analyzed the anti-angiogenic activity of decursin on retinal neovascularization. First, to investigate the anti-angiogenic activity of decursin on retinal neovascularization in OIR, we performed fluorescence angiography. On P17 in OIR, many neovascular tufts at the border of vascularized and nonvascularized retina were easily detected (Figure 3A), whereas they were significantly reduced in decursin-treated mice (Figure 3B).

Bottom Line: We evaluated the inhibitory effect of decursin on retinal neovascularization.Decursin significantly inhibited VEGF-induced proliferation, migration, and the formation of capillary-like networks of retinal endothelial cells in a dose-dependent manner.Moreover, the upregulation of glial fibrillary acidic protein expression was not detected in Mueller cells.

View Article: PubMed Central - PubMed

Affiliation: Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Department of Ophthalmology, College of Medicine, Seoul National University and Seoul Artificial Eye Center Clinical Research Institute, Seoul National University Hospital, Seoul 151-744, Korea.

ABSTRACT

Purpose: Pathologic angiogenesis in the retina leads to the catastrophic loss of vision. Retinopathy of prematurity (ROP), a vasoproliferative retinopathy, is a leading cause of blindness in children. We evaluated the inhibitory effect of decursin on retinal neovascularization.

Methods: Anti-angiogenic activity of decursin was evaluated by vascular endothelial growth factor (VEGF)-induced proliferation, migration, and in vitro tube formation assay of human retinal microvascular endothelial cells (HRMECs). We also used western blot analysis to assess inhibition of vascular endothelial growth factor receptor-2 (VEGFR-2) phosphorylation by decursin. After intravitreal injection of decursin in a mouse model of ROP, retinal neovascularization was examined by fluorescence angiography and vessel counting in cross-sections. The toxicity of decursin was evaluated through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in HRMECs as well as histologic and immunohistochemistry examination for glial fibrillary acidic protein in the retina.

Results: Decursin significantly inhibited VEGF-induced proliferation, migration, and the formation of capillary-like networks of retinal endothelial cells in a dose-dependent manner. Decursin inhibited VEGF-induced phosphorylation of VEGFR-2, blocking the VEGFR-2 signaling pathway. When intravitreously injected, decursin dramatically suppressed retinal neovascularization in a mouse model of ROP. Even in a high concentration, decursin never induced any structural or inflammatory changes to cells in retinal or vitreous layers. Moreover, the upregulation of glial fibrillary acidic protein expression was not detected in Mueller cells.

Conclusions: Our data suggest that decursin may be a potent anti-angiogenic agent targeting the VEGFR-2 signaling pathway, which significantly inhibits retinal neovascularization without retinal toxicity and may be applicable in various other vasoproliferative retinopathies as well.

Show MeSH
Related in: MedlinePlus