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目的 探讨阳离子脂质体介导的内皮抑素 (ES)基因转移对角膜新生血管的抑制作用。方法 新西兰大白兔 30只,角膜缝线法制作角膜新生血管模型,按随机数字表法随机分为A、B、C3组。A组:缝线后立即于上方近角膜缘处结膜下注射 150μl阳离子脂质体与ES重组质粒 (含质粒 20μg)的混合液;B组:同样部位注射 150μl阳离子脂质体与空白质粒(含质粒 20μg)的混合液;C组:同样部位注射 150μl生理盐水。分别于缝线后 3、7、14、21和 28d在裂隙灯显微镜下观察各组角膜新生血管的生长情况,并计算角膜新生血管的面积。通过免疫组化方法检测各时相角膜缘及角膜组织中ES目的基因的表达情况。结果 A组角膜新生血管的出现时间为 ( 6 85±0 69 )d,B组为(3 43±0 53)d,C组为(3 14±0 69)d, 3组间比较差异有统计学意义 (F=100 24, P<0 05 ),A组明显晚于其他两组;在缝线后 14、21、28d,A组角膜新生血管的面积均明显小于其他两组,差异有统计学意义(F=72 662, 75 601, 27 729; P均<0 05)。A组于各观察时相均可在上方角膜缘和角膜组织中发现ES阳性表达,以转染 3d时最为明显, 7d时阳性细胞着色有所变浅, 14d时阳性细胞数明显减少, 28d时只偶见阳性表达;各时相下方角膜及角膜缘组织均未见ES阳性表达。B、C两组各时相均未发现ES阳?
Objective To investigate the inhibitory effect of cationic liposome-mediated endostatin (ES) gene transfer on corneal neovascularization. Methods Thirty new-born New Zealand white rabbits were randomly divided into A, B and C3 groups according to random number table. Group A: 150μl cationic liposome and ES recombinant plasmid (containing 20μg plasmid) were injected subconjunctivally immediately after the suture into the corneal margin. Group B: 150μl cationic liposome and blank plasmid Plasmid 20 μg); Group C: 150 μl of physiological saline was injected at the same site. The growth of corneal neovascularization was observed at 3, 7, 14, 21 and 28 days after suture in slit lamp microscope, and the area of corneal neovascularization was calculated. Immunohistochemistry was used to detect the expression of ES gene in the limbal and corneal tissues. Results The incidence of corneal neovascularization was (6 85 ± 0 69) days in group A, (3 43 ± 0 53) days in group B and (3 14 ± 0 69) days in group C, respectively (F = 100 24, P <0 05), group A was significantly later than the other two groups; the area of corneal neovascularization in group A at 14,21,28 d after suture was significantly less than the other two groups, with statistical differences Significance (F = 72 662, 75 601, 27 729; P <0.05). In group A, ES positive expression was found in the upper limbal and corneal epithelium at each observation time, which was the most obvious at 3 days of transfection. At 7 days, the staining of positive cells became lighter and the number of positive cells decreased significantly at 14 days. Only occasional positive expression was observed. No ES positive expression was observed in corneal and limbal tissues in each phase. B, C two groups did not find any time positive ES Yang?