title: Inhibition of choroidal neovascularization by topical application of angiogenesis inhibitor vasostatin.
abstract: OBJECTIVE
  Choroidal neovascularization (CNV) is the leading cause of blindness in patients with age-related macular degeneration (AMD). This study evaluated the inhibitory effect of vasostatin (VS), an endogenous angiogenesis inhibitor, on CNV.
  
  
  METHODS
  Anti-angiogenic activity of VS was evaluated in vitro by migration and tube formation assays in human umbilical vein endothelial cells (HUVECs). CNV lesions were induced in Brown Norway rats by fundus argon laser photocoagulation. Beginning one day after CNV induction, rats were treated with eye drops containing 1 microg/ml VS in PBS buffer for three times daily for 20 days. The extent of CNV was examined by flat mount analysis on day 24 or by fundus fluorescein angiography (FAG) on days 21, 28, 35, and 42, respectively. CNV lesions and choroidal vascularity were evaluated by histological analysis. The spatial distribution of topically applied VS in rat eyes was evaluated by immunoblot analysis.
  
  
  RESULTS
  VS inhibited migration and tube formation in HUVECs. Flat mount analysis revealed that, after laser-induced photocoagulation, topical VS application for 20 days significantly reduced CNV lesions. Moreover, serial FAG analysis indicated that a 20 day VS treatment significantly reduced CNV lesions on all subsequent days. Histological analysis revealed attenuated lesions, intact Bruch's membrane, and reduced choroidal vascularity in VS-treated eyes. Finally immunoblot analysis reveled VS expression in choroids.
  
  
  CONCLUSIONS
  Topical VS application suppresses the progression of laser-induced CNV via angiogenesis inhibition and may constitute a therapeutic alternative for excessive neovascularization occurring with ocular diseases.
authors.affiliation: Department of Ophthalmology, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan.
authors.forename: Shwu-Jiuan
authors.initials: SJ
authors.lastname: Sheu
authors.forename: Youn-Shen
authors.initials: YS
authors.lastname: Bee
authors.forename: Yi-Ling
authors.initials: YL
authors.lastname: Ma
authors.forename: Guei-Sheung
authors.initials: GS
authors.lastname: Liu
authors.forename: Hsiu-Chen
authors.initials: HC
authors.lastname: Lin
authors.forename: Tse-Liang
authors.initials: TL
authors.lastname: Yeh
authors.forename: Jau-Chen
authors.initials: JC
authors.lastname: Liou
authors.forename: Ming-Hong
authors.initials: MH
authors.lastname: Tai
mesh_terms: D000287:Administration, Topical; D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D037282:Calreticulin / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects; D020256:Choroidal Neovascularization / Q000517:prevention & control*; D042783:Endothelial Cells / Q000187:drug effects / Q000473:pathology; D005451:Fluorescein Angiography; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting; D007834:Lasers; D018919:Neovascularization, Physiologic / Q000187:drug effects; D010446:Peptide Fragments / Q000494:pharmacology*; D051381:Rats; D011914:Rats, Inbred BN; D012160:Retina / Q000187:drug effects
chemical_list: D020533:Angiogenesis Inhibitors; D037282:Calreticulin; D010446:Peptide Fragments; C121986:vasostatin