中华眼底病杂志

中华眼底病杂志

大鼠非动脉炎性前部缺血性视神经病变模型建立与评价

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目的 建立大鼠非动脉炎性前部缺血性视神经病变(NAION)模型,定性、定量评价视网膜、视神经的损伤情况。 方法 采用随机数字表法将健康雄性Sprague-Dawley大鼠47只分为正常对照组、单纯激光组、NAION模型组,各有13、11、23只大鼠。取右眼为实验眼。NAION模型组采用孟加拉玫瑰红联合激光光动力方法建立模型。单纯激光组大鼠采用与NAION模型组相同的激光参数照射视盘,不注射光敏剂。正常对照组大鼠不作干预。建模后12 h及1、3、7、28 d,利用直接检眼镜观察大鼠视网膜及视盘情况。使用苏木精伊红染色及透射电子显微镜定性评价NAION模型视盘、视网膜组织形态改变。利用经上丘荧光金逆行标记技术及荧光显微镜照相,定量评价NAION大鼠视网膜神经节细胞(RGC)密度及存活率。 结果 NAION模型组大鼠于建模后3 d视盘水肿明显,视神经轴突水肿、部分髓鞘解体;建模后7 d,视盘水肿基本消退;建模后28 d,视神经萎缩,大量轴突消失伴明显胶质化改变,RGC明显减少。正常对照组、单纯激光组大鼠无上述视盘及视网膜形态改变。NAION模型组与正常对照组、单纯激光组大鼠右眼RGC密度比较,差异均有统计学意义(t=−14.142、−14.088,P=0.000、0.000)。NAION模型组与正常对照组、单纯激光组大鼠RGC存活率比较,差异均有统计学意义(t=−17.048、−16.667,P=0.000、0.000)。正常对照组与单纯激光组大鼠RGC密度、存活率比较,差异无统计学意义(t=0.050、0.348,P=0.961、0.731)。 结论 孟加拉玫瑰红联合激光光动力方法可引起大鼠视神经轴突损伤及RGC死亡,成功建立NAION模型。

Objective To establish and evaluate a rat model of nonarteritic anterior ischemic optic neuropathy (NAION). Methods The rats were randomly divided into control group (n=13), sham laser group (n=11) and NAION group (n=23). The right eye was set as the experimental eye. NAION model was induced by directly illuminating the optic nerve (ON) of the right eye with 532 nm green laser, after intravenous infusion with the photosensitizing agent Rose Bengal. Sham laser treatment consisted of illuminating the ON region with 532 nm laser without Rose Bengal injection. Rats in control group underwent no intervention. The appearance of optic disc was observed with funduscope at 12 hours, 1, 3, 7, 28 days post-illumination. The histologic changes in the retina and ON of the NAION model were evaluated qualitatively with hematoxylin and eosin (HE) staining and transmission electron microscopy. The retrograde-labeled retinal ganglion cells (RGC) were counted on photographs taken from retinal flat mounts in a masked fashion. Results The optic disc in NAION eyes were swollen 3 days after photodynamic treatment. HE-stained longitudinal ON sections of NAION revealed vacuolar degeneration on day 3 after induction. Besides, ultrastructural study showed axonal edema and collapsed sheaths in the ischemic optic nerve at the same time point after modeling. ON edema resolved 7 days after induction. The final results revealed optic disc atrophy, extensive axonal loss, severe glial scar, and RGC death in large numbers 4 weeks after modeling. There were no aforementioned manifestations in control and sham laser group. The RGC density of the right eyes was statistically significantly lower in NAION group than that in control group and in sham laser group (t=−14.142, −14.088; P=0.000, 0.000). The survival rate of RGC was statistically significantly lower in NAION group than in control group and in sham laser group (t=−17.048, −16.667; P=0.000, 0.000). There was no difference of RGC density and survival rate of RGC between control and sham laser group (t=0.050, 0.348; P=0.961, 0.731). Conclusion A rat model of NAION was established successfully by photodynamic treatments with Rose Bengal, which induce optic nerve damage and RGC death.

关键词: 视神经病变,缺血性; 视网膜神经节细胞; 疾病模型,动物

Key words: Optic neuropathy, ischemic; Retinal ganglion cells; Disease models, animal

引用本文: 王一玮, 陈婷, 马瑾, 钟勇. 大鼠非动脉炎性前部缺血性视神经病变模型建立与评价. 中华眼底病杂志, 2018, 34(1): 60-64. doi: 10.3760/cma.j.issn.1005-1015.2018.01.015 复制

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1. Miller NR. Current concepts in the diagnosis, pathogenesis, and management of nonarteritic anterior ischemic optic neuropathy[J]. J Neuroophthalmol, 2011, 31(2): 1-3. DOI: 10.1097/WNO.0b013e31821f955c.
2. Bernstein SL, Johnson MA, Miller NR. Nonarteritic anterior ischemic optic neuropathy (NAION) and its experimental models[J]. Prog Retin Eye Res, 2011, 30(3): 167-187. DOI: 10.1016/j.preteyeres.2011.02.003.
3. Bernstein SL, Guo Y, Kelman SE, et al. Functional and cellular responses in a novel rodent model of anterior ischemic optic neuropathy[J]. Invest Ophthalmol Vis Sci, 2003, 44(10): 4153-4162. DOI: 10.1167/iovs.03-0274.
4. 王润生, 王小娣, 吕沛霖, 等. 光动力诱导前部缺血性视神经病变的实验研究[J]. 中华眼底病杂志, 2008, 24(2): 90-94.Wang RS, Wang XD, Lyu PL, et al. Experimental study on photodynamic induced anterior ischemic optic neuropathy in rat animals[J]. Chin J Ocul Fundus Dis, 2008, 24(2): 90-94.
5. Ma J, Jiang L, Zhong Y, et al. Neuroprotective effect on retinal ganglion cells by transpupillary laser irradiation of the optic nerve head[J]. Neurosci Lett, 2010, 476(1): 3-8. DOI: 10.1016/j.neulet.2010.01.001.
6. Ma K, Xu L, Zhang H, et al. Effect of brimonidine on retinal ganglion cell survival in an optic nerve crush model[J]. Am J Ophthalmol, 2009, 147(2): 326-331. DOI: 10.1016/j.ajo.2008.08.005.
7. Sugiyama K, Gu ZB, Kawase C, et al. Optic nerve and peripapillary choroidal microvasculature of the rat eye[J]. Invest Ophthalmol Vis Sci, 1999, 40(13): 3084-3090.
8. Hayreh SS. Ischemic optic neuropathy[J]. Prog Retin Eye Res, 2009, 28(1): 34-62. DOI: 10.1016/j.preteyeres.2008.11.002.
9. Knox DL, Kerrison JB, Green WR. Histopathologic studies of ischemic optic neuropathy[J]. Trans Am Ophthalmol Soc, 2000, 98:203-222.
10. Zhang C, Guo Y, Slater BJ, et al. Axonal degeneration, regeneration and ganglion cell death in a rodent model of anterior ischemic optic neuropathy (rAION)[J]. Exp Eye Res, 2010, 91(2): 286-292. DOI: 10.1016/j.exer.2010.05.021.
11. Slater BJ, Mehrabian Z, Guo Y, et al. Rodent anterior ischemic optic neuropathy (rAION) induces regional retinal ganglion cell apoptosis with a unique temporal pattern[J]. Invest Ophthalmol Vis Sci, 2008, 49(8): 3671-3676. DOI: 10.1167/iovs.07-0504.
12. Bernstein SL, Mehrabyan Z, Guo Y, et al. Estrogen is not neuroprotective in a rodent model of optic nerve stroke[J]. Mol Vis, 2007, 13:1920-1925.
13. Schmid H, Renner M, Dick HB, et al. Loss of inner retinal neurons after retinal ischemia in rats[J]. Invest Ophthalmol Vis Sci, 2014, 55(4): 2777-2787. DOI: 10.1167/iovs.13-13372.
14. Vidal-Sanz M, Salinas-Navarro M, Nadal-Nicolas FM, et al. Understanding glaucomatous damage: anatomical and functional data from ocular hypertensive rodent retinas[J]. Prog Retin Eye Res, 2012, 31(1): 1-27. DOI: 10.1016/j.preteyeres.2011.08.001.
15. Sanchez-Migallon MC, Valiente-Soriano FJ, Nadal-Nicolas FM, et al. Apoptotic retinal ganglion cell death after optic nerve transection or crush in mice: delayed rgc loss with BDNF or a Caspase 3 inhibitor[J]. Invest Ophthalmol Vis Sci, 2016, 57(1): 81-93. DOI: 10.1167/iovs.15-17841.
16. Parrilla-Reverter G, Agudo M, Nadal-Nicolas F, et al. Time-course of the retinal nerve fibre layer degeneration after complete intra-orbital optic nerve transection or crush: a comparative study[J]. Vision Res, 2009, 49(23): 2808-2825. DOI: 10.1016/j.visres.2009.08.020.
17. Aktas Z, Gurelik G, Gocun PU, et al. Matrix metalloproteinase-9 expression in retinal ganglion cell layer and effect of topically applied brimonidine tartrate 0.2% therapy on this expression in an endothelin-1-induced optic nerve ischemia model[J]. Int Oph-thalmol, 2010, 30(3): 253-259. DOI: 10.1007/s10792-009-9316-9.