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Updated: May 12, 2026
Assessment of Vascular Regeneration in the CNS Using the Mouse Retina
Published on: June 23, 2014
Small interference RNA targeting vascular endothelial growth factor gene effectively attenuates retinal
Yi-chun Kong1, Bei Sun, Kan-xing Zhao
1Tianjin Eye Hospital; Tianjin Key Lab of Ophthalmology and Visual Science; Tianjin Eye Institute; Clinical College of Ophthalmology, Tianjin Medical University, Tianjin 300020, China (Email: kongyc1942@hotmail.com).
Background:
The mechanism of retinal neovascularization is not understood completely. Many growth factors are involved in the process of retinal neovascularization, such as vascular endothelial growth factor (VEGF) and pigment epithelium-deprived factor (PEDF), which are the representatives of angiogenic and antiangiogenic molecules respectively. Oxygen induced retinopathy (OIR) is a useful model to investigate retinal neovascularization. The present study was conducted to investigate the feasibility of small interference RNA (siRNA) targeting VEGF gene in attenuating oxygen induced retinopathy (OIR) by regulating VEGF to PEDF ratio (VEGF/PEDF).
Methods:
In vitro, cultured EOMA cells were transfected with VEGF-siRNA (psi-HI(TM)/EGFP/VEGF siRNA) and Lipofectamine(TM) 2000 for 24, 48, and 72 hours, respectively. Expression of VEGF mRNA was evaluated by real time polymerase chain reaction (PCR) and the level of VEGF protein was analyzed by Western blotting. In vivo, OIR model mice were established, the mice (C57BL/6J) received an intra-vitreal injection of 1 µl of mixture of psi-HI(TM)/EGFP/VEGF siRNA and Lipofectamine 2000. Expressions of retinal VEGF and PEDF protein were measured by Western blotting, retinal neovascularization was observed by fluorescein angiography, and quantified.
Results:
In vitro psi-HI(TM)/EGFP/VEGF siRNA treatment significantly reduced VEGF mRNA and protein expression. In vivo, with decreased VEGF and VEGF-PEDF ratio, significant attenuation of neovascular tufts, avascular regions, tortuous, and dilated blood vessels were observed in the interfered animals.
Conclusions:
VEGF plays an important role in OIR, and the transfection of VEGF-siRNA can effectively downregulate VEGF expression in vivo, accompanied by the downregulation of VEGF-PEDF ratio, and simultaneous attenuation of retinal neovascularization was also observed. These findings suggest that VEGF/PEDF may serve as a potential target in the treatment of retinal neovascularization and RNA interference targeting VEGF expression, which represents a possible therapeutic strategy.
Insights
Small interference RNA (siRNA) targeting vascular endothelial growth factor (VEGF) effectively reduced VEGF expression in oxygen-induced retinopathy (OIR) models. This therapeutic strategy attenuated retinal neovascularization by modulating the VEGF to PEDF ratio.
Area of Science:
- Ophthalmology
- Molecular Biology
- Genetics
Background:
- Retinal neovascularization mechanisms remain incompletely understood.
- Vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) are key regulators of angiogenesis.
- Oxygen-induced retinopathy (OIR) serves as a model for studying retinal neovascularization.
Purpose of the Study:
- To investigate the efficacy of VEGF-targeting small interference RNA (siRNA) in attenuating OIR.
- To evaluate the impact of VEGF-siRNA on the VEGF to PEDF ratio in OIR models.
Main Methods:
- In vitro: EOMA cells were transfected with VEGF-siRNA; VEGF mRNA and protein levels were assessed via real-time PCR and Western blotting.
- In vivo: OIR mice received intra-vitreal injections of VEGF-siRNA; retinal VEGF and PEDF protein levels were measured by Western blotting.
- Retinal neovascularization was visualized and quantified using fluorescein angiography.
Main Results:
- VEGF-siRNA significantly reduced VEGF mRNA and protein expression in vitro.
- In vivo, VEGF-siRNA treatment led to decreased VEGF levels and an altered VEGF/PEDF ratio.
- Significant attenuation of retinal neovascularization, including reduced neovascular tufts and improved vascular integrity, was observed.
Conclusions:
- VEGF plays a critical role in oxygen-induced retinopathy.
- VEGF-siRNA effectively downregulates VEGF expression in vivo, impacting the VEGF/PEDF ratio.
- Targeting VEGF with RNA interference presents a potential therapeutic strategy for retinal neovascularization.
