Studies on ocular neovascularization by DNA microarray and RNA interference

采用Smith等人的方法制备小鼠视网膜新生血管。采用affmetrix基因芯片MGU74 AV2检测小鼠感觉视网膜mRNA表达变化。在P15时，共有129个基因的上调或下调幅度超过对照组的两倍或一半。在上调的基因中，我们特别关注了βB2-crystallin。在新生血管丛周围的胶质细胞中检测到β b2 -晶体蛋白和a-肌动蛋白的免疫反应。给予β b2 -晶体蛋白特异性短干扰RNA，可抑制模型小鼠眼部新生血管的形成。mRNA和蛋白的表达也受到抑制。在玻璃体手术获得的人体组织中检测β b2 -晶体蛋白免疫反应性。将βB2-crystallin基因转染HEK293细胞，观察细胞功能。在转染细胞中，观察到细胞过程的伸长，发现β b2 -晶体蛋白和a-肌动蛋白在细胞膜下共定位。我们的研究结果表明β b2 -晶体蛋白在眼（视网膜）新生血管中起重要作用。

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.