血管内皮细胞生长抑制因子VEGI_(151)重组腺病毒对角膜新生血管的抑制作用

Effect of adenovirus mediated vascular endothelial growth inhibitor 151 on corneal neovascularization

目的研究以复制缺陷型重组腺病毒为载体的血管内皮细胞生长抑制因子VEGI151对角膜新生血管的抑制作用。方法利用腺病毒载体pCA13构建携带VEGI151基因的质粒,经293A细胞包装、扩增,WesternBlot检测病变细胞内基因的蛋白质表达,体外检测AdVEGI151对ECV304细胞的抑制作用,缝线诱导法建立炎症性兔角膜新生血管模型,给予体内基因治疗。结果VEGI151重组腺病毒在病变细胞内能成功表达相对分子质量为17300的蛋白,体外实验表明,可以强烈抑制静脉内皮细胞的生长,体内实验表明,重组腺病毒对炎症性角膜新生血管的生长具有抑制作用。结论以复制缺陷型重组腺病毒为载体的血管内皮细胞抑制因子VEGI151基因治疗角膜新生血管效果显著,为角膜新生血管的基因治疗提供了新的思路。

ObjectiveTo study the therapeutic effects of adenovirus mediated vascular endothelial cell growth inhibitor 151(VEGI_ 151 ) on corneal neovascularization(CNV).MethodsVEGI_ 151 gene was cloned into adenovirus vector and then packed in 293A cell to make Ad-VEGI_ 151 .The expressed protein was demonstrated by Western blot.The inhibitory effect of Ad-VEGI_ 151 on endothelial cells (ECV304) was detected in vitro.Inflammatory CNV models in rabbit were established and the therapeutic effects on CNV were observed.ResultsAd-VEGI_ 151 could express functional protein at the relative molecular weight 17300 .Growth of ECV304 cells was significantly suppressed at 24hours and the OD values in various time points were significantly decreased in Ad-VEGI_ 151 group in comparison with Ad-LacZ group and PBS group(P<0.01,P<0.01),and the area of inflammatory CNV was significantly reduced in Ad-VEGI_ 151 group_ compared with Ad-LacZ group and PBS group(P<0.01,P<0.01).ConclusionAd-VEGI_ 151 could inhibit the proliferation of CNV.It might be a potential route for treating CNV.