VEGI基因转移抑制角膜新生血管的实验研究(英文)

Inhibition of corneal neovascularization by vascular endothelia growth inhibitor gene

目的:探讨以阳离子脂质体介导重组人VEGI基因转移对角膜新生血管(corneal neovascularization,CNV)的抑制作用。方法:角膜缝线法制作兔CNV模型,用结膜下注射的方法将阳离子脂质体包裹的VEGI重组质粒(pcDNA4-VEGI)转染入兔角膜,裂隙灯显微镜下观察记录各组兔CNV长出的时间、长度和数量,并分别于基因转染后3,7,14及21d以免疫组织化学方法观察VEGI基因表达情况,观察其对CNV的抑制作用。结果:基因转染组CNV平均出现时间为6.3d,对照组分别为3.1,3.2,3.2d不等,差异有统计学意义(F=39.838,P<0.01);基因转染后3d,转染组实验兔未出现CNV,对照组已有部分兔眼出现CNV;转染后第7d,转染组实验兔的CNV纤细,累及钟点数局限于缝线周围,对照组兔的CNV最长为2.9mm,血管密集;转染后第14d,转染组兔CNV最长达4.0mm,对照组新生血管最长达6.4mm,累及钟点数为3.2个;各时间段基因转染组CNV长度、平均面积与对照组相比,差异均有统计学意义(q=17.386,P<0.01)。免疫组织化学显示角膜上皮、基质、新生血管管壁细胞的VEGI表达阳性。各实验指标与对照组比较,差异有统计学意义(均P<0.05)。结论:VEGI基因对CNV有抑制作用。

AIM：To evaluate the effect of EffecteneTM lipofectine mediated plasmids encoding human pcDNA4-vascular endothelia growth inhibitor（pcDNA4-VEGI） gene on corneal neovascularization（CNV）.METHODS：Forty New Zealand albino rabbits were sutured by 5-0 silk on the superior cornea to induce CNV and divided into 4 random teams,ten per each team：team A：transfected by pcDNA4-VEGI gene mediated by EffecteneTM lipofectine transfection;team B：by plasmid pcDNA4;team C：by EffecteneTM,and team D：by normal saline.Length and area of CNV were observed under slit lamp every day after transfection.Immunohistochemistry was performed to detect the expression of VEGI protein in corneas at day 3,7,14 and 21.RESULTS：1） Average occurrence of CNV was 6.3 days in team A,3.1 days in team B,3.2 days in team C,and 3.2 days in team D.Difference was significant between A and other teams（P0.01）;2） Length and average area of CNV in each period in team A was significantly different from those in team B,C and D（P0.01）;3） VEGI expressions were observed in epithelium,stroma,endothelium and the cliff of CNV in team A at 3 days after transfection by immunohistochemical staining.None VEGI positive cells were found in the control teams（team B,C and D） all the time.CONCLUSION：EffecteneTM lipofectine transfection technique can effectively transfect pcDNA4-VEGI gene into rabbit cornea and the length and CNV areas can be inhibited by VEGI gene.