【摘要】 目的　探讨脂质体介导endostatin基因体内转染对脉络膜新生血管 (choroidalneovascularization ,CNV)模型的抑制效应。方法　激光光凝方式建立brownnorway大鼠CNV模型 ,随机分为endostatin组 ,空质粒组 ,对照组 ;脂质体介导法分别导入重组endostatin基因真核表达载体pSecTagA hEndostatin或空载质粒pSecTagA。原位杂交、免疫组化和ELISA法检测endostatin基因mRNA转录及其蛋白表达 ;脉络膜血管平铺、FFA、CD10 5免疫组化观察对CNV的抑制效应。结果　Endo statin基因可有效转染视网膜、RPE及脉络膜并得到表达。转基因后 7、14d眼组织匀浆endostatin蛋白表达量为 ( 5 0 14±3 43 )ng /眼和 ( 3 1 5± 2 2 1)ng/眼 ;Endostatin组CNV面积、荧光渗漏程度、CD10 5阳性细胞表达量与空质粒组及对照组差别显著。结论　脂质体介导endostatin基因体内转染可以有效抑制CNV的形成。更多还原

【Abstract】 Objective To investigate the effects of liposome mediated intraocular gene transfection of endostatin on the inhibition of the development of choroidal neovascularization (CNV) in a rat model. Methods Experimental CNV model in Brown Norway rats was induced by laser photocagulation. The recombinant eukaryotic expression plasmid pSecTagA hEndostatin or control plasmid pSecTagA and liposome complexes were injected into the subretinal space of the model rats. In situ hybridization and immunohistochemical observation confirmed the presence of endostatin mRNA and protein expression two weeks after injection. Intraocular and serum levels of endostatin were measured by enzyme linked immunosorbent assay. The intensity of fluorescein leakage from the photocoagulated lesions was studied at 13 d after photocoagulation. The area of CNV was measured using high molecular weight FITC dextran (MW2×10 6) for high resolution angiography in RPE choroid sclera flat mounts. In addition, sections of CNV lesions were studied by light microscopy and endoglin (CD105) immunohistochemical evaluation. Results The retina, RPE, choroidal were infected by subretinal delivery of the pSecTag hEndostatin and expressed the endostatin. Two weeks after intraocular injection, the level of endostatin in the whole eye homogenates were (50 14±3 43) ng/eye and (31 5±2 21) ng/eye, respectively. Fluorescein leakage from the CNV lesions decreased significantly as compared with that in the control groups. The average area of CNV at the sites of the Bruch’s membrane rupture showed significant difference in eyes injected with endostatin as compared with that in the control eyes. Endothelial cells demonstrated strong immunoreactivity of CD105 in CNV lesions in the control eyes. Conclusion Liposome mediated endostatin gene transfection can significantly inhibit the development of CNV.更多还原