Inhibitory effects of rAAV2-pigment epithelial derived factor on oxygen-induced retinal neovascularization in mice

2010 
Objective To investigate the effects of recombinant adeno-associated virus type-2 (rAAV2) mediated delivery of pigment epithelium-derived factor (PEDF) on oxygen-induced retinal neovascularization (OIRNV) in mice. Methods A total of 22 C57/BL6 mice at the age of 3 days received intravitreal injections of 1 μl rAAV2-PEDF and rAAV2-EGFP into the left eyes (experimental group) and the right eyes (control group). All mice were put into the oxygen box right after the injection to induce the OIRNV model. 4 mice were sacrificed and PEDF protein in retina was measured by western blot at postnatal days 13 (P13). Twelve mice underwent retinal angiography with high molecular weight fluorescein-dextran,and another 6 mice were sacrificed for retinal lectin immunohistochemistry staining at P17. Absolute and relative non-perfusion areas of retinal neovascularization were analyzed by Image-Pro Plus 5.1 software.Results The expression level of PEDF protein was higher in the experimental group than that in the control group. The absolute non-perfusion area was (0. 96 + 0.22) mm2 in the experimental group and (1.96±0. 34) mm2 in the control group; the difference between the two groups was significant (t = -8. 554, P<0.01). The relative non-perfusion area was (8. 64 ± 1.52) % in the experimental group and (17. 27 ± 2. 98)% in the control group with a significant difference between the two groups (t = -8. 97, P<0. 01). The absolute area of retinal neovascularization was (0. 37 ± 0. 11) mm2 in the experimental group which was obviously higher than (1.26±0. 38) mm2 in the control group (t=-7. 8, P<0. 01); the relative areas in experimental and control groups was (3. 96 ± 0. 66) % and ( 11.45 ± 2. 06) %, respectively, whose difference is apparently (t=-8. 51, P<0. 01). The areas of retina neovascularization were (0. 11±0. 003)mm2 and (0.41±0.02)mm2 in the experimental and control groups, respectively, and the differencebetween the two groups was significant (t =- 5.14, P< 0. 01). Conclusions PEDF protein can stably express in the mice retina after rAAV2-PEDF transfetion, rAAV2-PEDF can decrease the retinal nonperfusion areas and inhibit the retinal neovascularization in OIRNV mice. Key words: Retinal neovascularization/prevention & control;  Cytokines;  Gene transfer techniques;  Transfeetion;  Animal experimentation
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