Zhou Lian-hong, Xing Yi-qiao, Chen Chun-lian, Wang Dao-wen
Department of Ophthalmology, Renmin Hospital of Wuhan University, Wuhan 430060, China.
Zhonghua Yan Ke Za Zhi. 2006 May;42(5):426-30.
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.
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.
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).
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.
探讨重组腺相关病毒介导的血管内皮生长因子反义基因（rAAV-aVEGF）转染是否能抑制大鼠角膜新生血管（CNV）的形成。
采用三质粒共转染法构建rAAV-aVEGF(165)和重组腺相关病毒介导的Lacz（rAAV-Lacz）。48只Sprague-Dawley（SD）大鼠随机分为两组。通过角膜中央碱性烧灼在体内诱导CNV。24只大鼠结膜下注射rAAV-aVEGF(165)（10(10) pfu/ml）；另一些大鼠结膜下注射rAAV-Lacz（10(10) pfu/ml）作为对照。通过5-溴-4-氯-3-吲哚-β-D-半乳糖苷（X-gal）免疫组化染色评估Lacz基因表达。烧灼后1至30天，通过形态学分析评估CNV，并通过免疫组化评估VEGF的表达。
2天后，对照组结膜囊中检测到21.36%±1.07%的Lacz基因表达。30天后，28.02%±1.16%的Lacz基因仍有表达。形态学分析和免疫组化表明rAAV介导的反义VEGF(165)显著抑制CNV。与注射rAAV-Lacz的对照组相比，烧灼后VEGF(165)蛋白减少（F = 1639.22，F = 2187.16，F = 719.17，P < 0.01）。
本研究表明rAAV-aVEGF(165)能充分抑制烧灼诱导的CNV，其作用与抑制VEGF产生有关。
