脂质体介导基因转染活体眼组织的研究

Experimental observation of gene transfer with liposome to murine ocular tissues by different routes of administration

目的 了解活体内不同途径应用脂质体转染外源基因在眼组织的分布情况。方法 将包有 β 半乳糖苷酶基因质粒的脂质体DOSPER ,分别采用玻璃体腔内注射、尾静脉注射、表面点药及球后注射到小鼠体内 ,2h ,1,2天、1,2 ,4周后用 β gal染色法观察 β 半乳糖苷酶基因在眼组织内的分布情况。 结果 玻璃体腔内注射、尾静脉注射、表面点药及球后注射 4组的小鼠眼球组织包括角膜、虹膜、睫状体、视网膜 ,均有 β 半乳糖苷酶基因表达 ,除表面点药组外 ,其他 3组小鼠脉络膜也有 β 半乳糖苷酶基因表达。用药 1天后开始出现 β 半乳糖苷酶基因阳性表达 ,持续达 1个月以上。 结论 脂质体能有效、稳定地转染外源基因到活体眼组织的角膜、虹膜、睫状体、脉络膜、视网膜。

ObjectiveTo compare different routes of administration on the ability of liposomes carrying an exogenous gene to introduce that gene into murine ocular tissues.MethodsPlasmid DNA(pGME 7Zf(+)) with beta galactosidase gene (DOSPER) was applied topically to eyes or injected into vitreous,tail vein,or retro orbital region of adult mice.Control and treated mice eyes were enucleated at 2?h,1?d and 2?d,1 week,2?and 4?weeks post injection or topical application.Gene expression was detected by enzymatic color reaction using beta gal stain as a substrate in frozen sections of enucleated eyes.ResultsCell positive for beta galactosidase expression showed blue staining of enzymatic color reaction in tissue sections.Liposome mediated gene transfer was detected in murine cornea,iris,ciliary body and retinal ganglion cells,photoreceptors,and retinal pigment epithelium using all four routes of administration Injection into vitreous,tail vein and retro orbital region allowed transfer of the gene into the choroid.The gene could not be transferred into murine choroid by topical application.Gene expression in murine ocular tissues was observed after 1?d following injection or topical application.The expression lasted at least 1?month.Blue staining indicating an enzymatic color reaction was not observed in all of control eyes.ConclusionEfficient and stable transfer of functional genes could be achieved by liposome delivery into the cornea,iris,ciliary body,choroid and retina of mice.