[Antisense vascular endothelial growth factor suppressed corneal neovascularization in rats].
OBJECTIVE
To explore whether recombinant adeno-associated virus mediated antisense vascular endothelial growth factor (rAAV-aVEGF) gene transfer inhibits the development of corneal neovascularization (CNV) in a rat model.


METHODS
rAAV-aVEGF(165) and recombinant adeno-associated virus mediated Lacz (rAAV-Lacz) were constructed by three-plasmid cotransfection methods. Forty-eight Sprague-Dawley (SD) rats were divided into two groups randomly. CNV were induced in vivo by alkaline cauterization of the central cornea. Twenty-four rats were injected with rAAV-aVEGF(165) (10(10) pfu/ml) into conjunctiva; another rats were injected with rAAV-Lacz (10(10) pfu/ml) into conjunctiva as controls. The Lacz gene expression was evaluated by 5-bromo-4-chloro-3-indolyl-beta-D-galactoside (X-gal) immunohistochemical staining. 1 to 30 days post-cautery, CNV was evaluated by morphometric analysis, and expression of VEGF was evaluated by immunohistochemistry.


RESULTS
Two days later, 21.36% +/- 1.07% Lacz gene expression was detected in conjunctival sac in control group. 30 d later, 28.02% +/- 1.16% Lacz gene remained expression. Morphometric analysis and immunohistochemistry demonstrated rAAV-mediated antisense VEGF(165) significantly inhibited CNV. The VEGF(165) protein was decreased post-cautery compared to control group injected with. rAAV-Lacz (F = 1639.22, F = 2187.16, F = 719.17, P < 0.01).


CONCLUSIONS
This study suggests that rAAV-aVEGF(165) can sufficiently inhibit the cautery-induced CNV, and the effect is associated with the inhibition of VEGF production.
Zhou
Lian-hong
LH
Department of Ophthalmology, Renmin Hospital of Wuhan University, Wuhan 430060, China. zlh681102@yahoo.com.cn
Xing
Yi-qiao
YQ
Chen
Chun-lian
CL
Wang
Dao-wen
DW
D000818:Animals; D003315:Cornea / Q000378:metabolism*; D016510:Corneal Neovascularization / Q000378:metabolism / Q000628:therapy*; D000229:Dependovirus / Q000235:genetics*; D005126:Eye Burns / Q000150:complications; D015316:Genetic Therapy; D005822:Genetic Vectors; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000235:genetics*
D020319:Oligodeoxyribonucleotides, Antisense; D042461:Vascular Endothelial Growth Factor A