Vaccinium myrtillus (Bilberry) Extracts Reduce Angiogenesis In Vitro and In Vivo

Department of Biofunctional Evaluation, Molecular Pharmacology, Gifu Pharmaceutical University, 5-6-1 Mitahora-higashi, Gifu 502-8585 and Wakasa Seikatsu Co. Ltd, 22 Naginataboko-cho, Shijo-Karasuma, Shimogyo-ku, Kyoto 600-8008, Japan.
Evidence-based Complementary and Alternative Medicine (Impact Factor: 1.88). 11/2007; 7(1):47-56. DOI: 10.1093/ecam/nem151
Source: PubMed


Vaccinium myrtillus (Bilberry) extracts (VME) were tested for effects on angiogenesis in vitro and in vivo. VME (0.3-30 µg ml(-1)) and GM6001 (0.1-100 µM; a matrix metalloproteinase inhibitor) concentration-dependently inhibited both tube formation and migration of human umbilical vein endothelial cells (HUVECs) induced by vascular endothelial growth factor-A (VEGF-A). In addition, VME inhibited VEGF-A-induced proliferation of HUVECs. VME inhibited VEGF-A-induced phosphorylations of extracellular signal-regulated kinase 1/2 (ERK 1/2) and serine/threonine protein kinase family protein kinase B (Akt), but not that of phospholipase Cγ (PLCγ). In an in vivo assay, intravitreal administration of VME inhibited the formation of neovascular tufts during oxygen-induced retinopathy in mice. Thus, VME inhibited angiogenesis both in vitro and in vivo, presumably by inhibiting the phosphorylations of ERK 1/2 and Akt. These findings indicate that VME may be effective against retinal diseases involving angiogenesis, providing it can reach the retina after its administration. Further investigations will be needed to clarify the major angiogenesis-modulating constituent(s) of VME.

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    • "Animal studies have demonstrated B-ext to be beneficial in preventing retinal inflammation and cataracts [15]. Our previous studies have shown a neuroprotective effect against retinal neuronal damage induced by N-methyl-d-aspartic acid in mice [16] and an inhibitory effect against angiogenesis in a mouse model of oxygen-induced retinopathy [17].Lingonberry is used in traditional medicine for the treatment of frequent urination, sore eyes, toothache, snow blindness, and thrush [18]. "
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    ABSTRACT: Blue light is a high-energy or short-wavelength visible light, which induces retinal diseases such as age-related macular degeneration and retinitis pigmentosa. Bilberry (Vaccinium myrtillus L.) and lingonberry (Vaccinium vitis-idaea) contain high amounts of polyphenols (anthocyanins, resveratrol, and proanthocyanidins) and thus confer health benefits. This study aimed to determine the protective effects and mechanism of action of bilberry extract (B-ext) and lingonberry extract (L-ext) and their active components against blue light-emitting diode (LED) light-induced retinal photoreceptor cell damage. Cultured murine photoreceptor (661 W) cells were exposed to blue LED light following treatment with B-ext, L-ext, or their constituents (cyanidin, delphinidin, malvidin, trans-resveratrol, and procyanidin B2). 661 W cell viability was assessed using a tetrazolium salt (WST-8) assay and Hoechst 33342 nuclear staining, and intracellular reactive oxygen species (ROS) production was determined using CM-H2DCFDA after blue LED light exposure. Activation of p38 mitogen-activated protein kinase (p38 MAPK), nuclear factor-kappa B (NF-κB), and LC3, an ubiquitin-like protein that is necessary for the formation of autophagosomes, were analyzed using Western blotting. Caspase-3/7 activation caused by blue LED light exposure in 661 W cells was determined using a caspase-3/7 assay kit. B-ext, L-ext, NAC, and their active components improved the viability of 661 W cells and inhibited the generation of intracellular ROS induced by blue LED light irradiation. Furthermore, B-ext and L-ext inhibited the activation of p38 MAPK and NF-κB induced by blue LED light exposure. Finally, B-ext, L-ext, and NAC inhibited caspase-3/7 activation and autophagy. These findings suggest that B-ext and L-ext containing high amounts of polyphenols exert protective effects against blue LED light-induced retinal photoreceptor cell damage mainly through inhibition of ROS production and activation of pro-apoptotic proteins.
    BMC Complementary and Alternative Medicine 04/2014; 14(1):120. DOI:10.1186/1472-6882-14-120 · 2.02 Impact Factor
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    • "Angiogenesis is the physiological process of forming new blood vessels from the preexisting vasculature, and it is a vital process during embryonic development. However, in adults angiogenesis is only observed in specific areas, such as the endometrium and ovarian follicle cells [1]. Angiogenesis also plays a key role in many diseases, including cancer, where it promotes tumor growth and metastasis [2]. "
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    ABSTRACT: Background Gleditsia sinensis thorns have been widely used in traditional Korean medicine for the treatment of several diseases, including obesity, thrombosis, and tumor-related diseases. The aim of the study is to determine the antiangiogenic effect of Gleditsia sinensis thorns in vitro and in vivo in a bid to evaluate its potential as an anticancer drug. Methods Ethanol extract of Gleditsia sinensis thorns (EEGS) were prepared and used for in vitro and in vivo assays. In vitro antiangiogenic effect of EEGS was determined in HUVEC primary cells by cell migration and tube formation assays. In vivo antiangiogenic effect of EEGS was determined by measuring vessel formation and vascular endothelial cells migrating into the implanted matrigels in nude mice. The angiogenesis-related proteins of which expression levels were altered by EEGS were identified by proteomic analysis. Results EEGS exerted a dose-dependent antiproliferative effect on HUVEC cells without significant cytotoxicity. Angiogenic properties, such as cell migration and tube formation, were significantly inhibited by EEGS in a dose-dependent manner. New vessel formation was also suppressed by EEGS, as determined by the directed in vivo angiogenesis assays in nude mice. EEGS reduced the expression of proangiogenic proteins, endothelin 1 and matrix metallopeptidase 2, in HUVEC cells. Conclusions Our findings suggest that EEGS can inhibit angiogenesis by down-regulating proangiogenic proteins, and therefore it should be considered as a potential anticancer drug targeting tumor-derived angiogenesis.
    BMC Complementary and Alternative Medicine 12/2012; 12(1):243. DOI:10.1186/1472-6882-12-243 · 2.02 Impact Factor
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    • "In the present study, VMA, delphinidin, cyanidin, malvidin and NAC all displayed both antiangiogenic and DPPH radical-scavenging effects (Figs 2–4; also see Matsunaga et al., 2007). Taken together, the above observations suggest that the inhibitory actions of VMA against angiogenesis may be due in part to the radical-scavenging effects of the constituent anthocyanins (including delphinidin, cyanidin and malvidin). "
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    ABSTRACT: The aim of this study was to examine the antiangiogenic properties and antioxidant activities (a) of the main anthocyanidins (delphinidin, cyanidin and malvidin) found as constituents in Vaccinium myrtillus (bilberry) anthocyanosides (VMA) and (b) of N-acetyl-L-cysteine (NAC). Each of these anthocyanidins concentration-dependently inhibited vascular endothelial growth factor (VEGF)-induced tube formation in a co-culture of human umbilical vein endothelial cells (HUVECs) and fibroblasts, the effect of each anthocyanidin being significant at 3 and/or 10 microM, while NAC significantly inhibited such tube formation at 1 microM (the only concentration tested). Moreover, each anthocyanidin (0.3-10 microM) and NAC (1-1000 microM) concentration-dependently scavenged the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. The inhibitory effects against angiogenesis were similar among the anthocyanidins, as were those against the DPPH radical. Moreover, their radical-scavenging effects were induced by concentrations that were at or below those that induced their antiangiogenic effects. These findings indicate that the inhibitory effect of VMA on angiogenesis may depend on those of its main constituent anthocyanidins (delphinidin, cyanidin and malvidin), presumably via antioxidant effects.
    Phytotherapy Research 01/2010; 24 Suppl 1(S1):S42-7. DOI:10.1002/ptr.2895 · 2.66 Impact Factor
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