中文摘要：

目的探讨早产儿视网膜病（ROP）差异表达基因，分析其功能通路，丰富ROP发病机制，为ROP防治提供理论依据。 方法获取引产胎儿眼球，分离培养视网膜微血管内皮细胞，按照培养基氯化钴（CoCl2）浓度（0、100、150、200、250、300、350 μmol/L）分为7组，并最终利用150 μmol/L CoCl2诱导缺氧制备ROP体外细胞模型。采用Ⅷ因子和CD31抗体荧光染色验证视网膜微血管内皮细胞，进行RNA提纯，基因芯片杂交，微阵列显著分析软件筛选表达差异基因，基因本体学分析软件研究差异表达基因的功能通路。 结果1.CoCl2≥150 μmol/L，细胞增殖活性开始下降（F＝21，P〈0.05）。2.150 μmol/L CoCl2组的二代和三代血管内皮细胞Ⅷ因子抗体染色胞核呈蓝色，胞质为绿色，CD31单克隆抗体染色的血管内皮细胞在胞质中可见红色荧光。无CoCl2组二代和三代血管内皮细胞Ⅷ因子和CD31抗体染色仅见胞核染色，但胞质未见染色。3.人胚视网膜微血管内皮细胞缺氧诱导后326个基因表达存在差异，198个基因表达上调，128个基因表达下调，上调基因表达约是下调基因的1.5倍。4.基因本体学分析软件提示差异基因与细胞缺氧、血管生成抑制、铁离子转运等10个生物过程通路明显相关。 结论CoCl2诱导视网膜微血管内皮细胞缺氧可制备ROP体外细胞模型，ROP存在差异表达基因，细胞缺氧、血管生成抑制、铁离子转运等功能通路可能与ROP发病机制相关。

英文摘要：

Objective To study the differentially expressed genes and analyze its functional pathways of reti- nopathy of prematurity（ROP） ,in order to discover the pathogenesis and provide the theoretical basis for the prevention and treatment of ROP. Methods Fetal eyeballs of induced labor were obtained and retinal mierovascular endothelial cells were isolated and cultured. The endothelial cells were divided into 7 groups according to the medium of cobalt chloride （COC12） concentration （0 μmol/L, 100 μmol/L, 150 μmol/L,200 μmol/L,250 μmol/L,300 μmol/L, 350 μmol/L）, and 150 μmol/L CoC12 was finally used to induce ROP model in vitro. Retinal microvascular endothelial cells were verified by adopting Ⅷ factor and CD31 antibody fluorescence staining. RNA purification, gone chip hybridiza- tion and signi-ficant analysis of microarrays were performed to screen differentially expressed genes. Genes functional pathways were studied by using gone ontology analysis software. Results （ 1 ） The proliferation activity of vascular en- dothelial cells decreased when CoC12 ≥150μmol/L（ F = 21 ,P 〈 0.05 ）. （ 2 ） In 150μmol/L CoC12 group, blue nucleus and green cytoplasm were visible in the second and the third generation vascular endothelial ceils stained by factor Ⅷ antibody, and red fluorescence could also be observed in the cytoplasm by means of CD31 monoclonal antibody staining. However, only blue nucleus was detected in the group without CoCl2. （3）There were 326 genes differently expressed in retinal micro vascular endothelial cells induced by CoCl2 in vitro, among whom, 198 genes were up -regulated and 128 genes were down - regulated. Up - regulated expression genes were 1.5 times more than those of the down - regulated genes. （4）Ten biological pathways including cell hypoxia, angiogenesis suppression and iron ion transport etc. may play important roles in ROP pathogenesis. Contusions Hypoxic retinal microvascular endothelial cells induced by CoCl2 can successfully be used to induce cel