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J. Lee, S. Lee, H. Koh, K.-H. Chung; Inhibitary Effect of Saxatilin on Cultured Porcine Choriocapillary Endothelium and Rat Choroidal Neovascularization. Invest. Ophthalmol. Vis. Sci. 2010;51(13):3355. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
to investigate the suppressive effect of saxatilin, disintegrin from snake venom, on adhesion, migration, and proliferation of choriocapillary endothelium, which plays a critical role in CNV, and to evaluate inhibitory effect on experimental choroidal neovascularization in rats
Primary porcine choriocapillary endothelial cells (CECs) were obtained and cultured. We evaluated inhibitary effect of saxatilin on adehesion, migration, proliferation on CECs. Also, we investigated effect of saxatilin on viability of CECs. CNV were induced by laser photocoagulation (532 nm wavelength, 150 mW, 60 µm, 50 ms) in both eyes of 30 adult Brown Norway rats, followed by intraviteal administration of single dose of saxatilin of 0 µg (n=10), 3 µg (n=10), or 15 µg (n=10).
Saxatilin inhibited the adhesion of CECs to fibronectin-coated plates and the migration through fibronectin-coated transwell membrane in a dose-dependent manner. Also, it completely suppressed the proliferation of CECs in 100 µg/ml concentration. Through trypan blue dye exclusion test and CCK-8, it showed no effect on cell viability at the concentration up to 500 µg/ml. On fluorescein angiogram 14 days after CNV induction, there was a dose-dependent decrease in fluorescein leakage of saxatilin-treated groups compared with control group (1.06 ± 0.08 vs. 0.74 ± 0.02, vs. 0.68 ± 0.01; p<0.05). Histologically, the thickness of the CNV lesions was significantly (p<0.05) reduced in eyes that received saxatilin (82.85 ± 1.29 vs. 58.18±1.55 vs. 46.02±2.47 µm). There was no noticeable change in retinal morphology, thickness and cell morphology.
Saxatilin effectively inhibited adhesion, migration and proliferation of porcine CECs, and suppressed CNV progression in rat model of laser induced CNV, suggesting that this disintegrin may be beneficial agent in the prevention and treatment of CNV.
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