The Research for Elucidation of Molecular Mechanism of Glaucomatous Optic Neuropathy and Development of Efficacious Treatment

将粘弹性溶液Healon-V*注入两只食蟹猴的一侧眼内，制作实验性急性青光眼模型眼。用Parkins压平眼压计测量眼压，两只青光眼的眼压维持在30 mm Hg以上，而对侧眼的眼压在15 mm Hg以下。眼压升高3d后，无痛处死猴子，摘除青光眼和对照眼，包括球后视神经。在双目显微镜下切除筛板前、后的筛板组织，提取并纯化总RNA。利用DNA微阵列、人类基因组U133 2.0阵列、Affimetrix(寡核苷酸阵列，单色，约39,000个转录产物)检测眼压升高后上调/下调的RNA，并用Atlas-Image 2.01进行分析。在信号对数比为2.0和信号对数比为2.0的条件下，选择了90个上调基因；在信号对数比为-2.0的条件下，选择了62个下调基因。上调的基因包括细胞骨架(profilin 1、血栓反应蛋白2、Protocadherin 8、CD44抗原、BMP-2)、信号转导(G蛋白)、细胞外基质(IV型、VI型、XV型)、免疫反应(鸟氨酸结合蛋白2)、转录因子(v-fos、JunB)、转运蛋白(铜蓝蛋白)、TIMP-1等，下调的基因包括细胞骨架(少突胶质细胞糖蛋白)、免疫反应(髓鞘碱性蛋白)、转录因子(少突胶质细胞转录因子1)、转化生长因子-2等。眼压升高急性期组织重塑相关基因表达无明显增加或减少。

Visco-elastic solution Healon-V* was intra-camerally injected in the unilateral eye of two cynomolgus monkeys, and experimental acute glaucoma model eyes were produced. The intraocular pressure (IOP) was measured by Parkins' applanation tonometer, and the IOP of the two glaucomatous eyes were confirmed to be sustained over 30mmHg, while those of contralateral control eyes were under 15mmHg. After the IOP elevation of 3 days, the monkeys were painlessly sacrificed, and the glaucomatous and control eyes were enucleated including retrobulbar optic nerve. The lamina cribrosa tissue including pre-laminar and post-laminar portion were excised under binocular microscope, and the total RNA was extracted and purified. DNA microarray, Human Genome U133 2.0 Array, Affimetrix (Oligo-array, single color, approximately 39,000 transcriptional products) were utilized to detect up/down regulated RNA after IOP elevation, and the data was analyzed with Atlas-Image 2.01. Ninety up-regulated genes were selected under the condition of "detection : present-present" and "signal log ratio > 2.0", and 62 down-regulated genes were selected under the condition of "signal log ratio <-2.0". Up-regulated genes included cytoskeleton (profilin 1, thrombospondin 2, protocadherin 8, CD44 antigen, BMP-2), signal transduction (G protein), extracellular matrix (collagen types IV, VI, XV), immune response (guanylate binding protein 2), transcription factor (v-fos, junB), transport (ceruloplasmin), TIMP-1, etc. Down-regulated genes included cytoskeleton (oligodendrocyte myelin glycoprotein), immune response (myelin basic protein), transcription factor (oligodendrocyte transcription factor 1), TGF-2, etc. Meanwhile, there was no significant increase or decrease in the gene expression related to tissue remodeling in the acute phase of IOP elevation.