Studies on ocular neovascularization by DNA microarray and RNA interference

DNA微阵列和RNA干扰对眼部新生血管形成的研究

• 批准号: 16390496
• 负责人: YOSHIMURA Nagahisa
• 金额: $ 8.96万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390496/
• 关键词: retinopathy of prematurity; diabetic retinopathy; retinal neovascularization; crystalline; DNA microarray; siRNA

Retinal neovascularization in mice retina was produced by the method of Smith et al. The sensory retina was obtained from the model and mRNA expression changes were analyzed by Affimetrix Gene Chip MGU74 AV2. A total of 129 genes were found to be up- or down-regulated by more than twice or half of the control values in the model at P15. Among the up-regulated genes, we paid special attention to βB2-crystallin. βB2-crystallin and a-actin immunoreactivities were detected in glial cells that surrounded neovascular tufts. Short interfering RNA specific to βB2-crystallin were given to the model mice and ocular neovascularization was inhibited by the treatment. mRNA and protein expression were also inhibited by the treatment. βB2-crystallin immunoreactivities were detected in surgically obtained human tissues obtained by vitreous surgery. βB2-crystallin gene was transfected to HEK293 cells and cellular function was analyzed. In transfected cells, elongation of cellular process was observed and βB2-crystallin and a-actin were found to be co-localized underneath the cellular membrane. Our results showed that βB2-crystallin plays an important role in ocular (retinal) neovascularization.

采用Smith等的方法建立小鼠视网膜新生血管模型，取感觉视网膜，应用Affiliation Gene Chip MGU 74 AV 2分析其mRNA表达变化。共发现129个基因在P15时上调或下调超过模型中对照值的两倍或一半。在上调表达的基因中，我们特别关注了βB2-晶状体蛋白。βB2-晶体蛋白和α-肌动蛋白免疫反应在新生血管丛周围的胶质细胞中检测到。对模型小鼠给予βB2-晶状体蛋白特异性的短干扰RNA，治疗可抑制眼部新生血管的形成。mRNA和蛋白质表达也受到处理的抑制。在玻璃体手术获得的手术获得的人体组织中检测到βB2-晶状体蛋白免疫反应性。将βB2-晶状体蛋白基因转染HEK 293细胞，分析细胞功能。在转染的细胞中，观察到细胞突起的延长，并且发现βB2-晶体蛋白和α-肌动蛋白共定位于细胞膜下。我们的研究结果表明，βB2-晶状体蛋白在眼（视网膜）新生血管中起重要作用。