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Lu Liu, Bin Mo, Jian Jiao, Yankun Yue, Qingjun Lu, Wu Liu; Experimental study on transfecting Lentivirus-mediated pigment epithelium derived growth factor-green and fluorescent protein into normal rat retina and laser-induced choroidal neovascularization model. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4978.
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1.To observe the expression and location of the pigment epithelium derived growth factor-green fluorescent protein after transfecting Lentivirus-mediated fluorescent protein (PEDF-GFP-LV) into the retina of the normal BN rat.2.To further investigate the influence to choroidal neovascularization (CNV) model after the silencing of PEDF, and disscuss the relationship between PEDF and CNV. And to explore the appropriate administration of gene therapy.
1.Normal BN rats were given intravitreous injections of PEDF-GFP-LV or control lentivirus (NC-GFP-LV). Localization of GFP was evaluated by fluorescence microscopy, and the expression of PEDF was assessed by real-time polymerase chain reaction and western blot. 2.The experimental CNV models were randomly assigned to be intravitreally injected with NC-GFP-LV and PEDF-GFP-LV immediately after photocoagulation. On day 3, 7, 14 and 28 after laser photocoagulation, the CNV tissue was assessed qualitatively and quantitatively by FFA, OCT, histopathological sections and choroidal flatmounts.
1.Three days after transfecting PEDF-GFP-LV and NC-GFP-LV into BN rat eyes, green fluorescence was observed in the RPE layer under fluorescence microscopy and lasted 4 weeks. RT-PCR and western blot results showed that the expression of PEDF of the rat retina was significantly enhanced compared with the control group.2.Injection of PEDF-GFP-LV into rat eyes with CNV results in much lower fluorescein leakage than in eyes injected with NV-GFP-LV. The results of FFA, OCT, histopathological sections and choroidal flatmounts showed that the maximum thickness and areas of the CNV tissue was much smaller than the control group.
1.PEDF-GFP gene can be transfected into the retina and RPE cells effectively mediated by Lentivirus and their expression can maintain 4 weeks after transfection. 2. This study suggest that intraocular expression of PEDF-GFP-LV may provide a new treatment approach for ocular neovascularization. It can be expected to be used in the gene therapy for retinal and choroidal diseases.
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