巩膜静脉烧灼模型中青光眼样特征的验证。

Validation of glaucoma-like features in the rat episcleral vein cauterization model.

机构信息

Key Laboratory of Vision Loss and Restoration, Ministry of Education; Department of Ophthalmology, Peking University People's Hospital, Beijing 100044, China; State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, Guangdong 510060, China.

State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, Guangdong 510060, China.

出版信息

Chin Med J (Engl). 2014;127(2):359-64.

PMID:24438629

Abstract

BACKGROUND

Glaucoma, an irreversible optic nerve neuropathy, always results in blindness. This study aimed to evaluate glaucoma-like features in the rat episcleral vein cauterization (EVC) model by multiple in vivo and in vitro evidences.

METHODS

Wistar rat was used in this study. The elevated intraocular pressure (IOP) was induced by cauterization of three episcleral veins. IOP was monitored with Tono-Pen XL tonometer. Time-dependent changes to the neuronal retinal layers were quantified by Fourier domain-optical coherence tomography. The function of retina was evaluated by electroretinogram (ERG). Survival of retinal ganglion cells (RGCs) was quantified by retrograde labeling. Histology study was performed with retinal sections stained with hematoxylin-eosin, glial fibrillary acidic protein, and neuronal nuclear antigen. Retina and aqueous humor protein were extracted and cytotoxic protein tumor necrosis factor alpha (TNF-α) and alpha-2 macroglobulin (α2m) were measured with Western blotting.

RESULTS

EVC is a relatively facile intervention, with low failure rates (<5%). After surgical intervention, chronic mild IOP elevation (about 1.6-fold over normal, P < 0.05) was induced for at least 6 weeks without requiring a second intervention. High IOP causes chronic and progressive loss of RGCs (averaging about 4% per week), progressive thinning of neuronal retinal layers (3-5 μm per week), and reduction of a- and b-wave in ERG. EVC method can also induce glial cell activation and alterations of inflammation proteins, such as TNF-α and α2m.

CONCLUSION

EVC method can establish a robust, reliable, economic and highly reproducible glaucomatous animal model.

摘要

背景

青光眼是一种不可逆转的视神经病变，最终会导致失明。本研究旨在通过多种体内和体外证据评估大鼠巩膜静脉烧灼（EVC）模型中的青光眼特征。

方法

本研究使用 Wistar 大鼠。通过烧灼三条巩膜静脉来诱导升高的眼内压（IOP）。使用 Tono-Pen XL 眼压计监测 IOP。通过傅里叶域光学相干断层扫描定量评估神经元视网膜层的时间依赖性变化。通过视网膜电图（ERG）评估视网膜功能。通过逆行标记定量视网膜神经节细胞（RGC）的存活。用苏木精-伊红、胶质纤维酸性蛋白和神经元核抗原染色视网膜切片进行组织学研究。提取视网膜和房水蛋白，并用 Western blot 测量细胞毒性蛋白肿瘤坏死因子-α（TNF-α）和α2 巨球蛋白（α2m）。

结果

EVC 是一种相对简单的干预措施，失败率较低（<5%）。手术后，至少 6 周内会引起慢性轻度眼压升高（约为正常眼压的 1.6 倍，P<0.05），无需再次干预。高眼压会导致 RGC 慢性进行性丧失（平均每周约 4%），神经元视网膜层逐渐变薄（每周 3-5μm），ERG 的 a-和 b-波减少。EVC 方法还可以诱导神经胶质细胞激活和炎症蛋白改变，如 TNF-α和α2m。