title: Rosmarinic Acid Suppresses Subconjunctival Neovascularization in Experimental Glaucoma Surgery.
abstract: OBJECTIVE
  Excessive subconjunctival scarring is associated with increased angiogenesis and leads to filtration failure in glaucoma surgery. In this study, we describe that rosmarinic acid (RA) has anti-angiogenic activity during wound healing in a rabbit model of glaucoma surgery.
  
  
  METHODS
  Forty New Zealand rabbits underwent an experimental trabeculectomy and were randomly allocated into two treatment groups: RA group - treated with subconjunctival injections of 0.1 ml RA (15 mg/ml; n = 20) - and control group - treated with subconjunctival injections of 0.1 ml balanced salt solution (n = 20). The in vivo effect of RA was investigated after 5 and 15 d by measuring the intraocular pressure (IOP; with Tonopen) and bleb area and vascularity (using the Moorfields Bleb Grading System). Vascularization was also studied by counting histological blood vessels and by immunohistochemistry of vascular endothelial growth factor (VEGF) at the surgical site and by quantification of vessels in chicken's chorioallantoic membrane (CAM), treated with AR 500 μg/ml for 48 h.
  
  
  RESULTS
  On the fifth day, eyes of RA group displayed higher bleb area (3.6 ± 0.2 versus 1.8 ± 0.2; p = 0.004) and lower vascularity (3.0 ± 0.5 versus 4.0 ± 0.4; p = 0.009) than controls; however, difference in IOP reduction was not significant (-1.4 ± 0.3 versus -0.8 ± 0.3 mmHg; p = 0.226). Proportion of vessels/field (4.6 ± 0.5 versus 10.4 ± 0.9; p = 0.008) and VEGF immunostaining (15,347 ± 3788 versus 31,043 ± 3230; p = 0.019) also declined with RA treatment. However, at the 15th day, none of the parameters were different between the groups, except for vessels/field proportion (5.4 ± 1.0 versus 10.6 ± 1.6; p = 0.035). CAM exposed to AR inhibited vascularization (-45.67 ± 4.74%; p < 0.001).
  
  
  CONCLUSIONS
  These data indicate RA has a short-term anti-angiogenic effect and could be a potential modulator of neovascularization during subconjunctival healing at glaucoma filtration surgical sites.
authors.affiliation: a Department of Ophthalmology .
authors.forename: Juliana de Lucena Martins
authors.initials: Jde L
authors.lastname: Ferreira
authors.affiliation: b Department of Pathology , School of Medicine of Ribeirão Preto, University of São Paulo , Ribeirão Preto , Brazil and.
authors.forename: Fernando
authors.initials: F
authors.lastname: Chahud
authors.affiliation: b Department of Pathology , School of Medicine of Ribeirão Preto, University of São Paulo , Ribeirão Preto , Brazil and.
authors.forename: Leandra N
authors.initials: LN
authors.lastname: Ramalho
authors.affiliation: a Department of Ophthalmology .
authors.forename: Carolina Maria
authors.initials: CM
authors.lastname: Modulo
authors.affiliation: c Faculty of Pharmacy , Federal University of Minas Gerais , Belo Horizonte , Brazil.
authors.forename: Lorena Carla
authors.initials: LC
authors.lastname: Vieira
authors.affiliation: a Department of Ophthalmology .
authors.forename: Peter Sol
authors.initials: PS
authors.lastname: Reinach
authors.affiliation: a Department of Ophthalmology .
authors.forename: Maria de Lourdes Veronese
authors.initials: Mde L
authors.lastname: Rodrigues
authors.affiliation: c Faculty of Pharmacy , Federal University of Minas Gerais , Belo Horizonte , Brazil.
authors.forename: Armando Silva
authors.initials: AS
authors.lastname: Cunha
authors.affiliation: a Department of Ophthalmology .
authors.forename: Jayter Silva
authors.initials: JS
authors.lastname: Paula
mesh_terms: D020533:Angiogenesis Inhibitors / Q000627:therapeutic use*; D000818:Animals; D000975:Antioxidants / Q000627:therapeutic use*; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply; D002934:Cinnamates / Q000627:therapeutic use*; D003228:Conjunctiva / Q000098:blood supply* / Q000378:metabolism; D053630:Depsides / Q000627:therapeutic use*; D004195:Disease Models, Animal; D005260:Female; D005901:Glaucoma / Q000378:metabolism / Q000503:physiopathology / Q000601:surgery*; D007150:Immunohistochemistry; D056965:Injections, Intraocular; D007429:Intraocular Pressure / Q000502:physiology; D009389:Neovascularization, Pathologic / Q000188:drug therapy*; D011817:Rabbits; D014130:Trabeculectomy; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D000097123:Rosmarinic Acid
chemical_list: D020533:Angiogenesis Inhibitors; D000975:Antioxidants; D002934:Cinnamates; D053630:Depsides; D042461:Vascular Endothelial Growth Factor A; D000097123:Rosmarinic Acid
keywords: Filtration surgery; glaucoma; neovascularization; rosmarinic acid; wound healing