目的:通过聚乳酸-聚乙醇酸共聚物(PLGA)纳米粒子载体介导p27kip1基因转染视网膜色素上皮(retinal pigmentepithelium,RPE)细胞,观察其对视网膜脱离(retinadetachment,RD)的兔眼RPE增殖的抑制情况。方法:制备p27kip1基因纳米粒子,并进行基因含量的测定。然后制备RD动物模型,应用免疫组织化学法检测PCNA蛋白的表达,蛋白质印迹法检测p27kip1蛋白在各视网膜细胞的表达。结果:p27kip1基因治疗组于7,14,28d的PCNA蛋白表达阳性率较对照组明显低,差异具有统计学意义(P<0.05),表明PCNA蛋白阳性表达受到p27kip1基因纳米粒子的明显抑制。且基因转染组3,7,14d时p27kip1蛋白表达较对照组明显增多,差异具有统计学意义(P<0.05)。结论:p27kip1基因有可能作为一个目的基因,借助新兴的基因载体纳米粒子用于抑制RPE细胞增殖的基因治疗。

AIM: Through p27kip1-gene mediated by the emerging polylactic acid-polyglycolic acid copolymer(PLGA) nanoparticles transfecting retinal pigment epithelium(RPE),to observe the inhibition effect of RPE proliferation of the retinal detachment(RD) rabbit eye·METHODS: Preparation of p27kip1 gene nanoparticle was made and determination of gene content was measured.Animal grouping and preparation of RD animal model were made.Experimental data were analyzed using paired t-test and independent samples t-test method,there were significant differences(P<0.05)·RESULTS: At 7,14,28 days,PCNA protein expression of p27kip1 gene therapy group was significantly lower than the control group,and the difference was statistically significant(P<0.05).This indicated that PCNA protein expression were significantly inhibited by p27kip1 gene nanoparticles.At the same time,in the gene transfection group,p27kip1 protein expression was significantly increased compared with the control group at 3,7,14 days,the difference was significant(P<0.05)·CONCLUSION: p27kip1 gene may be used as a target gene,and through the emerging gene nanoparticles may be used for the gene therapy about inhibition of RPE cell proliferation.

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