Molecular and Cellular PharmacologyInhibition of vascular endothelial growth factor (VEGF)-induced endothelial proliferation, arterial relaxation, vascular permeability and angiogenesis by dobesilate
Introduction
Angiogenesis is the formation of new blood vessels from the existing vasculature and is vital for reproduction, development and repair (Folkman, 1995). However, excessive angiogenesis is associated with a large number of pathological processes, including growth, invasion and metastasis in malignancies (Hanahan and Folkman, 1996). Angiogenesis is finely regulated by pro-angiogenic and anti-angiogenic stimuli. When this regulation is disbalanced, either favoring pro-angiogenic factors or limiting anti-angiogenic factors, pathological angiogenesis occurs. Vascular endothelial growth factor (VEGF) is, together with fibroblast growth factors (FGF), one of the most important promoters of angiogenesis. In addition to stimulating endothelial cell proliferation and migration (Byrne et al., 2005), VEGF is known to cause vasodilation (Liu et al., 2002) and to increase vascular permeability (Bates et al., 2002). All these actions contribute to the pathophysiological processes related to VEGF over-activity.
Increased production of VEGF has been observed in diseases characterized by excessive angiogenesis, namely age-related macular degeneration (Bhutto et al., 2006), diabetic retinopathy (Aiello et al., 1994), rheumatoid arthritis (Lee et al., 2001), psoriasis (Detmar et al., 1994), rosacea (Smith et al., 2007) and inflammatory bowel disease (Tolstanova et al., 2009). Elevated VEGF production by solid tumors is associated with increased tumor vascularization, metastasis, chemoresistance and poor prognosis in cancer processes (Foekens et al., 2001, Karayiannakis et al., 2002). Therapies directed to selectively neutralize VEGF have been proved to be effective in treating cancer (Shih and Lindley, 2006) and ocular neovascularization (Lynch and Cheng, 2007). However, the anti-angiogenic agents available nowadays, including antibodies or aptamers directed against VEGF or its receptor and tyrosine kinase inhibitors, present inconveniences for the administration, are expensive, and most importantly, are associated with serious side effects (Johnson et al., 2004).
2,5-dihydroxybenzene sulfonate (DHBS; dobesilate) is a small synthetic molecule that has been widely used in treating diabetic retinopathy and chronic venous insufficiency with a good safety profile (Allain et al., 2004, Ribeiro et al., 2006). However, the mechanism of action of this drug has not been completely elucidated. Recently, it has been demonstrated that DHBS binds to the heparin-binding domain of FGF-1 reducing its activity (Fernandez et al., 2010) and DHBS has been proved to be effective for the treatment of rosacea (Cuevas and Arrazola, 2005b) and psoriasis (Cuevas and Arrazola, 2005a), two clinical manifestations related to excessive angiogenesis and over-expression of VEGF.
The aim of the present study was to evaluate, using both in vitro and in vivo approaches, the ability of DHBS to antagonize the stimulatory actions of VEGF on endothelial cell proliferation, vasodilation, vascular permeability and angiogenesis.
Section snippets
Human endothelial cell culture
Freshly delivered human umbilical cord specimens were collected from the Hospital de Getafe in Madrid, as approved by the Hospital Ethics Committee. Human umbilical vein endothelial cells (HUVEC) were obtained by treatment with 0.1% type II collagenase and characterized by both morphology and positive staining for factor VIII, as previously described (Peiro et al., 2007). Cells were routinely cultured in M199 medium supplemented with 2% endothelial cell growth supplement (ECGS) and 20% fetal
Effects of DHBS on VEGF-induced proliferation of human endothelial cells
In the presence of 1% fetal calf serum, the treatment of cultured HUVEC for 72 h with VEGF (10 ng/ml) resulted in increased cell proliferation (Fig. 1A). The simultaneous addition of DHBS concentration-dependently antagonized the VEGF-induced proliferation of HUVEC. This effect was significant with DHBS from 50 μM concentration, while 100 μM DHBS completely abolished the proliferative stimulus driven by VEGF (Fig. 1A). In contrast, DHBS, at 50 or 100 μM, did not significantly modify the
Discussion
DHBS has been used for more than 30 years in the treatment of diabetic retinopathy, showing consistent efficacy (Ribeiro et al., 2006). Nevertheless, the mechanism responsible for this effect has been poorly elucidated. DHBS has been considered as an angioprotective agent and has been shown to decrease blood viscosity and vascular permeability in patients with diabetic retinopathy (Berthet et al., 1999). In addition, it has been reported to possess antioxidant capacity that could account for
Conclusion
The present study demonstrates that DHBS is an efficient in vitro and in vivo inhibitor of VEGF signaling pathway and VEGF-induced actions, including endothelial cell proliferation, arterial vasodilatation, vascular permeability and angiogenesis. These findings provide a rationale for explaining, at least in part, its therapeutic properties. The dual inhibitory effects of DHBS on VEGF and FGF activities makes this compound a promising candidate for treating a variety of deleterious conditions
Acknowledgments
This work was supported by grants from Action Medicines SL, Madrid, Spain, and from Ministerio de Ciencia e Innovación, Spain (SAF2008-01291; SAF2008-00942). Tania Romacho is the recipient of a fellowship from Ministerio de Educación y Ciencia.
References (57)
- et al.
Heparin regulates vascular endothelial growth factor165-dependent mitogenic activity, tube formation, and its receptor phosphorylation of human endothelial cells. Comparison of the effects of heparin and modified heparins
J. Biol. Chem.
(2005) - et al.
Pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor (VEGF) in aged human choroid and eyes with age-related macular degeneration
Exp. Eye Res.
(2006) - et al.
Neuropilins in neoplasms: expression, regulation, and function
Exp. Cell Res.
(2006) - et al.
Angioprotective action of calcium dobesilate against reactive oxygen species-induced capillary permeability in the rat
Eur. J. Pharmacol.
(1998) - et al.
Drug resistance by evasion of antiangiogenic targeting of VEGF signaling in late-stage pancreatic islet tumors
Cancer Cell
(2005) - et al.
VEGF-receptor signal transduction
Trends Biochem. Sci.
(2003) - et al.
Antiglioma effects of a new low molecular mass, inhibitor of fibroblast growth factor
Neurosci. Lett.
(2011) - et al.
Gentisic acid, a compound associated with plant defense and a metabolite of aspirin, heads a new class of in vivo fibroblast growth factor inhibitors
J. Biol. Chem.
(2010) - et al.
Leads for development of new naphthalenesulfonate derivatives with enhanced antiangiogenic activity: crystal structure of acidic fibroblast growth factor in complex with 5-amino-2-naphthalene sulfonate
J. Biol. Chem.
(2003) - et al.
Human brain-derived acidic and basic fibroblast growth factors: amino terminal sequences and specific mitogenic activities
Biochem. Biophys. Res. Commun.
(1986)
The binding of vascular endothelial growth factor to its receptors is dependent on cell surface-associated heparin-like molecules
J. Biol. Chem.
Patterns and emerging mechanisms of the angiogenic switch during tumorigenesis
Cell
Inhibition of choroidal angiogenesis by calcium dobesilate in normal Wistar and diabetic GK rats
Eur. J. Pharmacol.
Vascular endothelial growth factor-mediated, endothelium-dependent relaxation in human internal mammary artery
Ann. Thorac. Surg.
Nifedipine, losartan and captopril effects on hyperplasia of vascular smooth muscle from Ren-2 transgenic rats
Eur. J. Pharmacol.
1H-NMR assignment and solution structure of human acidic fibroblast growth factor activated by inositol hexasulfate
J. Mol. Biol.
Reduction of retinal albumin leakage by the antioxidant calcium dobesilate in streptozotocin-diabetic rats
Eur. J. Pharmacol.
Bevacizumab: an angiogenesis inhibitor for the treatment of solid malignancies
Clin. Ther.
Antiangiogenic and antitumor activities of peptide inhibiting the vascular endothelial growth factor binding to neuropilin-1
Life Sci.
Vasorelaxation induced by vascular endothelial growth factor in the human internal mammary artery and radial artery
Vascul. Pharmacol.
Antiangiogenic antithrombin blocks the heparan sulfate-dependent binding of proangiogenic growth factors to their endothelial cell receptors: evidence for differential binding of antiangiogenic and anticoagulant forms of antithrombin to proangiogenic heparan sulfate domains
J. Biol. Chem.
Vascular endothelial growth factor in ocular fluid of patients with diabetic retinopathy and other retinal disorders
N. Engl. J. Med.
Safety of calcium dobesilate in chronic venous disease, diabetic retinopathy and haemorrhoids
Drug Saf.
Impairment of endothelium-dependent relaxation by increasing percentages of glycosylated human hemoglobin. Possible mechanisms involved
Hypertension
Regulation of microvascular permeability by vascular endothelial growth factors
J. Anat.
Calcium dobesilate: pharmacological profile related to its use in diabetic retinopathy
Int. J. Clin. Pract.
Angiogenic and cell survival functions of vascular endothelial growth factor (VEGF)
J. Cell. Mol. Med.
Amino acid and cDNA sequences of a vascular endothelial cell mitogen that is homologous to platelet-derived growth factor
Proc. Natl. Acad. Sci. U. S. A.
Cited by (41)
Comprehensive strategy of conduit guidance combined with VEGF producing Schwann cells accelerates peripheral nerve repair
2021, Bioactive MaterialsCitation Excerpt :These growth factors may perform many functions: protecting the survive of motor and sensory neurons, and stimulating the neurons or glia cells to proliferate and migrate in an autocrine loop model [21]. On the other hand, VEGF-A secreted by the Schwann cells loaded in HSPS-SC(VEGF) conduits could bind to VEGF receptor (VEGFR2) to induce proliferation, adhesion and migration of both vascular endothelial cells and Schwann cells, thereby enhancing the activation of downstream angiogenic signaling pathway [42]. In our study, no significant difference of VEGFR2 protein expression was shown among the five groups, while the expression levels of p-VEGFR2 in HSPS-SC(VEGF) group was significantly higher than that in the HSPS-VEGF group, implying that VEGF-A proteins secreted by Schwann cells were bioactive and promoted the phosphorylation of VEGFR2 protein.
Calcium Dobesilate and Micro-vascular diseases
2019, Life SciencesCitation Excerpt :Reducing platelet aggregation by inhibiting prostaglandin and reduce erythrocyte aggregation and viscosity of suspension [19]. Down-regulating the expression of vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF) to inhibit vascular endothelial cell proliferation [20–22]. DR is an important cause of blindness in the working age population and is a major socio-economic problem [23,24].
Haemostatic agent etamsylate in vitro and in vivo antagonizes anti-coagulant activity of heparin
2018, European Journal of PharmacologyCitation Excerpt :This idea is supported by the reported ability of dobesilate (DHPS anion is identical for both dobesilate and etamsylate) to bind FGF at heparin-binding domain and avoiding the adequate interaction of FGF with heparin or heparin-like molecules (Fernández et al., 2010). In this sense, etamsylate could potentially interfere with any factor binding to heparin through a heparin-binding domain, as suggested by the ability of dobesilate to antagonize biological activities of an isoform of vascular endothelium growth factor (VEGF) possessing a heparin-binding domain (Angulo et al., 2011). On the other hand, the potentiation of heparin-induced vasodilation at high concentration of etamsylate is not surprising since heparin-induced vasodilation is know to be endothelium-dependent (Tangphao et al., 1999; Tasatargil et al., 2005; Paredes-Gamero et al., 2010) and dobesilate has been shown to enhance endothelium-dependent relaxation (Ruiz et al., 1997; Ruiz and Tejerina, 1998; Angulo et al., 2003a, 2003b) and improve endothelial cell function (Suschek et al., 1997; Leal et al., 2010).