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

将粘弹性溶液Healon-V*注射于两只食蟹猴单侧眼内，制备实验性急性青光眼模型眼。采用帕金斯眼压计测量眼压，证实2只青光眼的眼压均维持在30mmHg以上，而对侧对照眼的眼压均在15mmHg以下。IOP升高3天后，无痛处死，青光眼和对照眼去核，包括球后视神经。在双眼显微镜下切除筛板组织，包括层前和层后部分，提取总RNA并纯化。利用DNA微阵列、Human Genome U133 2.0 Array、affmetrix （Oligo-array，单色，约39,000个转录产物）检测IOP升高后的上调/下调RNA，并用Atlas-Image 2.01软件对数据进行分析。在“检测：present-present”和“信号对数比> 2.0”条件下选择上调基因90个，在“信号对数比<-2.0”条件下选择下调基因62个。上调的基因包括细胞骨架（profilin 1、血小板反应蛋白2、原钙粘蛋白8、CD44抗原、BMP-2）、信号转导（G蛋白）、细胞外基质（IV、VI、XV型胶原）、免疫应答（鸟苷酸结合蛋白2）、转录因子（v-fos、junB）、转运（铜蓝蛋白）、TIMP-1等。下调基因包括细胞骨架（少突胶质细胞髓鞘糖蛋白）、免疫应答（髓鞘碱性蛋白）、转录因子（少突胶质细胞转录因子1）、TGF-2等。同时，IOP升高急性期组织重塑相关基因表达无明显升高或降低。

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.