Wang Hong, Wang Bing
Department of Ophthalmology, the Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Int J Ophthalmol. 2010;3(4):295-8. doi: 10.3980/j.issn.2222-3959.2010.04.04. Epub 2010 Dec 18.
To evaluate the effect of Effectene™ lipofectine mediated plasmids encoding human pcDNA(4)-vascular endothelia growth inhibitor (pcDNA(4)-VEGI) gene on corneal neovascularization (CNV).
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 pcDNA(4)-VEGI gene mediated by Effectene(™) lipofectine transfection; team B: by plasmid pcDNA(4); team C: by Effectene(™), 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.
Effectene(™) lipofectine transfection technique can effectively transfect pcDNA(4)-VEGI gene into rabbit cornea and the length and CNV areas can be inhibited by VEGI gene.
评估Effectene™脂质体介导的编码人pcDNA(4)-血管内皮生长抑制因子(pcDNA(4)-VEGI)基因的质粒对角膜新生血管(CNV)的影响。
40只新西兰白兔在角膜上方用5-0丝线缝合以诱导CNV,随机分为4组,每组10只:A组:用Effectene(™)脂质体转染介导的pcDNA(4)-VEGI基因转染;B组:用质粒pcDNA(4);C组:用Effectene(™);D组:用生理盐水。转染后每天在裂隙灯下观察CNV的长度和面积。在第3、7、14和21天进行免疫组织化学检测角膜中VEGI蛋白的表达。
1)A组CNV平均出现时间为6.3天,B组为3.1天,C组为3.2天,D组为3.2天。A组与其他组之间差异有统计学意义(P<0.01);2)A组各时期CNV的长度和平均面积与B、C、D组相比有显著差异(P<0.01);3)转染后3天,A组通过免疫组织化学染色在上皮、基质、内皮和CNV边缘观察到VEGI表达。对照组(B、C和D组)始终未发现VEGI阳性细胞。
Effectene(™)脂质体转染技术可有效将pcDNA(4)-VEGI基因转染到兔角膜中,且VEGI基因可抑制CNV的长度和面积。