University Eye Hospital Freiburg, Killianstrasse 5, 79106 Freiburg im Breisgau, Germany.
Exp Eye Res. 2012 Nov;104:7-14. doi: 10.1016/j.exer.2012.08.011. Epub 2012 Sep 7.
The main drawbacks of currently described pressure induced glaucoma animal models are, that intraocular pressure (IOP) either rises slowly, leading to a heterogeneous onset of glaucoma in the treated animals or that IOP normalizes before significant damage occurs, necessitating re-treatment. Furthermore, a variable magnitude of IOP increase often results when particles are introduced into the anterior chamber. In order to develop a simple and reproducible rat glaucoma model with sustained IOP elevation after a single treatment we induced occlusion of the chamber angle by anterior chamber paracentesis and subsequent laser coagulation of the limbal area with 35, 40 or 45 laser burns. Right eyes served as controls. IOP was measured three times weekly using TonoLab rebound tonometry in awake animals. After four weeks, retinal tissue was harvested and processed for whole mount preparation. The number of prelabeled, fluorogold-positive retinal ganglion cells (RGCs) was analyzed under a fluorescence microscope. The eyes were further analyzed histologically. Results are expressed as means and standard deviation. Amplitude and duration of the IOP elevation increased with the number of laser burns. Two weeks after 35, 40 or 45 translimbal laser burns the IOP difference between treated and control eye was 7.5 ± 5, 14 ± 8 or 19 ± 9 mmHg, respectively; the RGC density/mm(2) 28 days after treatment was 1488 ± 238 for control eyes (n = 31) and 1514 ± 287 (n = 10), 955 ± 378 (n = 10) or 447 ± 350 (n = 11) for the respective laser groups. Mean IOP of all control eyes over the observation period was 12.4 ± 0.8 mmHg. The chamber angle showed pigment accumulation in the trabecular meshwork of all laser groups and confluent peripheral anterior synechia after 40 and 45 laser burns. Histologic examination of the retina revealed increasing glia activation in a pressure dependant manner. In this study, >91% of laser treated rats developed secondary glaucoma with sustained IOP elevation for at least 2 weeks. The amount of IOP elevation and RGC loss correspond with the number of laser burns applied. This relatively high success rate after a single procedure may constitutes an advantage over established glaucoma models, as this decreases the risk of complications (e.g. corneal decompensation, intraocular bleeding or inflammation) and, thus, improves the outcome.
目前描述的压力诱导型青光眼动物模型的主要缺点是,眼内压(IOP)升高缓慢,导致治疗动物的青光眼发病时间不一致,或者 IOP 在发生明显损伤之前恢复正常,需要再次治疗。此外,当将颗粒引入前房时,通常会导致 IOP 升高幅度不同。为了开发一种简单且可重复的大鼠青光眼模型,我们通过前房穿刺术使房角闭塞,然后用 35、40 或 45 个激光灼伤对角膜缘区进行激光凝固,从而使 IOP 在单次治疗后持续升高。右眼作为对照。在清醒动物中每周使用 TonoLab 回弹眼压计测量三次 IOP。四周后,采集视网膜组织并进行全层铺片准备。在荧光显微镜下分析预先标记的荧光金阳性视网膜神经节细胞(RGC)的数量。进一步对眼睛进行组织学分析。结果表示为平均值和标准差。IOP 升高的幅度和持续时间随激光灼伤的数量而增加。在进行 35、40 或 45 次经角膜激光灼伤后的两周,治疗眼与对照眼之间的 IOP 差异分别为 7.5 ± 5、14 ± 8 或 19 ± 9mmHg;治疗后 28 天的 RGC 密度/mm(2)分别为对照组(n = 31)的 1514 ± 287、激光组(n = 10)的 955 ± 378、激光组(n = 10)的 447 ± 350。所有对照眼在观察期间的平均 IOP 为 12.4 ± 0.8mmHg。在所有激光组中,前房角小梁网均有色素沉着,40 和 45 激光灼伤后出现广泛的周边前粘连。视网膜组织学检查显示,胶质细胞激活程度与压力呈依赖性增加。在这项研究中,超过 91%的激光治疗大鼠发展为继发性青光眼,其 IOP 升高至少持续 2 周。IOP 升高幅度和 RGC 损失与应用的激光灼伤数量相对应。单次手术后的高成功率可能是该模型相对于现有青光眼模型的优势,因为这降低了并发症的风险(例如角膜失代偿、眼内出血或炎症),从而改善了结果。