反义VEGF对视网膜新生血管的干预研究  

Vascular endothelial growth factor antisense inhibiting experimental retinal neovascularization

作　　者：曹晖[1] 许迅[1] 樊莹[1] 王方[1] 张皙[1] 

机构地区：[1]上海交通大学附属第一人民医院眼科,200080

出　　处：《眼科研究》2004年第3期256-259,共4页Chinese Ophthalmic Research

摘　　要：目的　将反义VEGF质粒用脂质体包裹注入视网膜新生血管模型小鼠眼中 ,观察其对新生血管的抑制作用。方法　将出生 7d的C5 7BL/6J小鼠 44只放入高氧环境中饲养 ,另外 8只于空气环境中饲养。 5d后出高氧环境 ,随机取3 6只分成 3组。将质粒与脂质体以 1∶5 (W/V)的比例混合 ,室温下静置 3 0min。大剂量组玻璃体腔内注入质粒PCR3 .1/Anti VEGF12 10 0 85 μg;小剂量组玻璃体腔内注入质粒PCR3 .1/Anti VEGF12 10 0 3 8μg;PCR3 .1组玻璃体腔内注入质粒PCR3 .10 0 5 3 μg,之后空气环境饲养 5d。解剖镜下视网膜铺片观察视网膜血管的分布情况 ;组织切片任意选取不包括视乳头的 2 0张切片 ,计数突出内界膜位于视网膜表面的细胞核数 ;VEGF免疫组化染色阳性细胞。结果　视网膜铺片原中周部血管紊乱分布部位 ,血管分布均匀。而对照组 (注射PCR3 .1)则无变化。内皮细胞计数小剂量组、大剂量组较高氧组、对照组显著减少 ,在P17的VEGF在毛细血管的血管内皮细胞的表达均有显著降低 ,但程度上有差异。结论　反义VEGF质粒对于视网膜新生血管的产生有抑制作用 ,在一定程度上减少了视网膜新生血管的产生 ,对血管内皮细胞表达VEGF有抑制作用。Objective To investigate the effect of vascular endothelial growth factor(VEGF) on retinal neovascularization,and determine the inhibitory effect of lipofectin-mediated antisense VEGF expression plasmid on experimental retinal neovascularization. Methods Forty-four of 52 one-week-old neonatal C57BL/6J mice were exposed to 75%±2% oxygen with the flux 1.5 L/min.The concentration of oxygen was measured 5 times per day and the temperature was controlled at 23℃±2℃.The others 8 mice were maintained in normal air condition.Recombined plasmid pCR3.1 endcoding antisense VEGF121 was extracted and purified by using QIAGEN EndoFree TM Plasmid Maxi Kit.The lipofectin-enwrapped pCR3.1/antiVEGF121 was injected into vitreous of exposed mice at high or low does of 0.085 μg or 0.038 μg respectively.Injection of 0.053 μg pCR3.1 served as the nagetive control.The inhibitory effect of VEGF on neovascularization was observed 5 days after treatment,during which atmospheric air environment was maintained. Results After treatment with pCR3.1/Anti-VEGF121,retinal avascula area was observed in exposed group in comparison with control group and in low dose group compared with high dose group.The numbers of endothelium cells decreased significantly in high dose group compared with control group or low dose group.Expression of VEGF was significantly decreased in both treated group and high dose group. Conclusion Intraocular injection of lipofectin-enwrapped antisense VEGF can inhibit retinal neovascularization and decrease the VEGF expression of retina tissue in dose-dependent mannor.

关 键 词：视网膜 新生血管 血管内皮生长因子 反义基因治疗 

分 类 号：R774[医药卫生—眼科]