Purpose: To investigate whether vector-based vascular endothelial growth factor 165 (VEGF)(165) targeted siRNA expression system (pSilencer(siVEGF)) could inhibit VEGF(165) expression in vitro and suppresses retinal neovascularization in the murine model of oxygen-induced retinopathy.
Methods: pSilencer(siVEGF), from which siRNA targeting VEGF(165) could be generated, was constructed and transfected to human umbilical vein endothelial cells. Then the level of VEGF isoforms in cultured cells was measured by RT-PCR and ELISA. Intravitreal injection of pSilencer(siVEGF) was performed in mice with ischemic retinopathy. Retinal neovascularization was evaluated by angiography using fluorescein-labeled dextran and quantitated histologically. The levels of VEGF(164), which is equivalent to human VEGF(165) in murine retinas were determined by RT-PCR and western immunoblotting.
Results: Expression of VEGF(165) in cultured cells was greatly curtailed by pSilencer(siVEGF) under both normoxia and hypoxia conditions. However, the other isoforms, VEGF(189) and VEGF(121), were expressed to a similar degree regardless of whether pSilencer(siVEGF) was administered. Based on angiography and histological analysis, retinal neovascularization in the eyes treated with pSilencer(siVEGF) were significantly reduced compared to the control eyes. Furthermore, the VEGF(164) levels in the murine retinas were suppressed by pSilencer(siVEGF).
Conclusions: Retinal neovascularization in the murine model was significantly attenuated by pSilencer(siVEGF) through decreasing VEGF(164) levels in the retinas. pSilencer(siVEGF) seems to be a potential therapeutic tool for ischemic-induced retinal diseases.
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Surv Ophthalmol
January 2025
School of Medicine, Vita-Salute San Raffaele University, Milan, Italy; Division of head and neck, Ophthalmology Unit, IRCCS San Raffaele Scientific Institute, Milan, Italy. Electronic address:
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Eye Clinic, Department of Biomedical and Clinical Sciences, Ospedale Luigi Sacco, University of Milan, Milan, Italy.
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Surv Ophthalmol
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Department of Biomedical Sciences, Humanitas University, Via Rita Levi Montalcini 4, 20072 Pieve Emanuele-Milan, Italy; Department of Ophthalmology, Eye Unit Humanitas Gavazzeni-Castelli, via Mazzini 11, Bergamo, Italy.
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Ophthalmology Department, Peking University People's Hospital, Beijing, China.
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Mol Pharm
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Key Laboratory of Chemical Biology (Ministry of Education), NMPA Key Laboratory for Quality Research and Evaluation of Carbohydrate-based Medicine, Department of Pharmacology, School of Pharmaceutical Sciences, Shandong University, 44 Wenhua West Road, Jinan 250012, China.
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