视神经损伤后的视网膜质子磁共振波谱检测和神经保护的研究

发布时间：2018-04-16 23:03

本文选题：视神经损伤 + 谷氨酸　；参考：《华中科技大学》2011年博士论文

【摘要】：目的利用质子磁共振波谱(1H magnetic resonance spectroscopy,1H-MRS)研究视神经横切(optic nerve transection, ONT)大鼠视网膜代谢物质的改变。方法成年SD大鼠双侧上丘注射2%荧光金逆行标记视网膜神经节细胞(retinal ganglion cells, RGCs)。荧光金标记后三天,将大鼠单侧视神经横切离断作为实验组。对照组大鼠未行视神经横切。视神经横切后2周,处死大鼠,分离视网膜。行视网膜铺片观察RGCs的平均密度。利用质子磁共振波谱研究大鼠视网膜提取物代谢物质的变化。免疫组织化学染色观察视网膜谷氨酸(glutamate,Glu),γ氨基丁酸(y-aminobutyric acid, GABA)和胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)的表达。结果视神经横切组与对照组相比,视网膜GABA、N-乙酰天门冬氨酸(N-acetylaspartate,NAA)、牛磺酸(taurine, Tau)、肌酸(creatine, Cr)浓度显著下降,丙氨酸(alanine, Ala)浓度显著升高。Glu、谷氨酰胺(glutamine, Gln)和Glx(Glu+Gln)浓度无统计学差异。对照组RGCs平均密度为2249±87/mm2,视神经横切组RGCs平均密度为320±56/mm2。与对照组相比,视神经横切组视网膜Glu和GABA免疫表达减弱,GFAP免疫表达显著增强。结论视神经横切后的RGCs死亡不仅与谷氨酸能和γ氨基丁酸能系统循环失衡有关,而且同时伴随其他的代谢物质平衡失调。质子磁共振波谱为我们提供了一个检测视网膜代谢变化的有效方法,这些代谢物质的变化为研究视神经横切后的RGCs死亡机制和神经保护提供了重要的依据。目的观察金丝桃素对视神经损伤大鼠视网膜神经节细胞存活的作用。方法将24只Sprague-Dawly (SD)大鼠随机分成正常对照组,单纯夹伤组,金丝桃素治疗组和生理盐水对照组。每组12只眼。大鼠双侧上丘注射2%荧光金逆行标记视网膜神经节细胞,7d后对除正常对照组以外其他各组进行球后视神经钳夹处理,同时玻璃体内分别注入金丝桃素或生理盐水5μL,14d后进行视网膜铺片和视网膜神经节细胞的计数。结果视神经夹伤后14d存活的视网膜神经节细胞数量显著减少。单纯夹伤组视网膜神经节细胞存活率为50%,生理盐水对照组视网膜神经节细胞存活率为52%,金丝桃素治疗组视网膜神经节细胞存活率为68%。金丝桃素治疗组相比单纯夹伤组和生理盐水对照组,存活的视网膜神经节细胞明显要多(P0.05)。结论玻璃体内注射金丝桃素不仅无毒副作用,而且能减少大鼠视神经夹伤后诱发的视网膜神经节细胞的死亡,对视网膜神经节细胞有保护作用。金丝桃素可能为视神经保护的新药研究带来新的思路。目的观察视神经夹伤大鼠晶状体损伤对视网膜神经节细胞存活和轴突再生的影响。方法将SD大鼠随机分成正常组、对照组和处理组。正常组不做任何处理。对照组行单纯双眼视神经钳夹。处理组视神经钳夹后,双眼玻璃体注射5μL生理盐水,依据注射方式的不同分为晶状体损伤组和晶状体未损伤组。7d后大鼠双侧上丘注射荧光金,14d时行视网膜铺片和冰冻切片,进行视网膜神经节细胞计数。并用免疫组织化学检测视网膜生长相关蛋白43(growth associated protein-43, GAP-43)和胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)的表达。结果正常组和对照组的视网膜神经节细胞计数分别为1987±154个·mm-2和908±123个·mm-2,处理组中的晶状体损伤组和晶状体未损伤组的视网膜神经节细胞计数分别为1206±134个·mm-2和800±112个·mm-2。晶状体损伤组存活的视网膜神经节细胞数比对照组和晶状体未损伤组多,差异有统计学意义(P0.05)。免疫组织化学显示晶状体损伤组视网膜GAP-43和GFAP的表达亦比对照组和晶状体未损伤组增强。结论晶状体损伤能促进视神经钳夹伤大鼠视网膜神经节细胞的存活,并增强视网膜GAP-43和GFAP的表达,促进轴突的再生。目的探讨视神经炎患者的视野、视觉诱发电位、眼底荧光血管造影和影像学等检查表现及临床治疗效果。方法100例视神经炎患者,分别行视野、视觉诱发电位、眼底荧光血管造影和影像学检查,并给予糖皮质激素、B族维生素、营养神经类药物、血管扩张剂及复方樟柳碱等综合治疗,观察各项检查表现及临床治疗效果。结果大多数视神经炎患者找不到明显发病诱因,视野、视觉诱发电位和眼底荧光血管造影有相应的非特异性异常表现,影像学检查特别是磁共振有助于发现隐匿的异常病灶,有重要的临床价值。经综合治疗后,85例视力提高,15例视力不变或下降。结论联合各项检查手段及时诊断及采取综合治疗是提高视神经炎患者疗效和预后的关键。
[Abstract]:Objective to study the changes of retinal metabolites in rat optic nerve transection (optic nerve transection, ONT) by means of 1H magnetic resonance spectroscopy (1H-MRS).
Methods adult SD rats with bilateral SC injection of 2% Fluorogold retrograde labeled retinal ganglion cells (retinal ganglion cells, RGCs). After three days of fluorescent gold labeling, the rats with unilateral optic nerve transection severed as the experimental group. The rats in control group without optic nerve transection. 2 weeks after optic nerve transection, death rat, isolated retina. The average density of retinal flatmount to observe the RGCs. Using the changes of metabolism of proton magnetic resonance spectroscopy study of retinal extract in rats. Immunohistochemical staining was used to observe the retinal glutamate (glutamate, Glu), gamma aminobutyric acid (Y-aminobutyric, acid, GABA) and glial fibrillary acidic protein (glial fibrillary acidic protein. The expression of GFAP).
Results the optic nerve transection group compared with control group, retinal GABA, N- acetylaspartate (N-acetylaspartate, NAA), taurine (taurine, Tau), creatine (creatine, Cr) were significantly decreased (alanine, Ala), alanine concentration significantly increased.Glu, glutamine (glutamine, Gln and Glx (Glu+Gln)) there was no significant difference in the control group RGCs concentration. The average density was 2249 + 87/mm2, optic nerve transection group average RGCs density was 320 + 56/mm2. compared with the control group, the optic nerve section group of retinal Glu and GABA immune expression decreased, GFAP expression was significantly enhanced.
Conclusion optic nerve after transection of the RGCs death not only with glutamatergic and GABAergic system cycle imbalance, and at the same time with other metabolic disorders. The material balance of proton magnetic resonance spectroscopy provides a useful method for the detection of retinal metabolic changes in the US, provides an important basis for the changes of these metabolites for optic nerve transection after studying the RGCs mechanism of death and neuroprotection.
Objective To observe the effect of hypericin on the survival of retinal ganglion cells in rats with optic nerve injury.
Methods 24 Sprague-Dawly (SD) rats were randomly divided into normal control group, simple injury group, hypericin treatment group and saline control group. Each group consisted of 12 eyes. Bilateral superior colliculus injection of 2% Fluorogold retrograde labeled retinal ganglion cells after 7d, except the normal control group to other groups of ball the visual nerve, and vitreous body were injected hypericin or saline 5 L, 14d after counting the retinas and retinal ganglion cells.
The results of optic nerve injury 14d survival of retinal ganglion cell numbers decreased significantly. The simple crush group of retinal ganglion cell survival rate was 50%, the saline control group of retinal ganglion cell survival rate was 52%, hypericin treatment group of retinal ganglion cell survival rate was 68%. hypericin treatment group compared with the injury group and normal the saline control group, the survival of retinal ganglion cells was significantly higher (P0.05).
Conclusion intravitreal injection of hypericin has no toxic side effect, and can reduce the death of rat retinal ganglion cells induced by optic nerve injury, has a protective effect on retinal ganglion cells. Hypericin may bring a new way of thinking for the research of new drugs of optic nerve protection.
Objective To observe the effect of lens injury on retinal ganglion cell survival and axonal regeneration in rats with optic nerve clamp injury.
Methods SD rats were randomly divided into normal group, control group and treatment group. The normal group without any treatment. The control group underwent bilateral optic nerve optic nerve. Treatment group after injection, eyes vitreous body 5 L normal saline, according to different injection methods for the damage of lens group and lens injury group without.7d after bilateral superior colliculus injection of fluorogold, 14d underwent retinal flatmount and frozen sections of retinal ganglion cell count. And immunohistochemical detection of retinal growth associated protein 43 (growth associated, protein-43, GAP-43) and glial fibrillary acidic protein (glial fibrillary acidic protein, GFAP) expression.
Results retinal ganglion cell count in normal group and control group were 1987 + 154 and 908 + 123 - mm-2 - mm-2, retinal ganglion cell count in the treatment group of lens injury group and lens without injury group were 1206 + 134 and 800 + 112 - mm-2 - mm-2. lens injury group the survival of retinal ganglion cell number compared with the control group and the non lens injury group, the difference was statistically significant (P0.05). Immunohistochemistry showed that the expression of GAP-43 in retina and lens injury group GFAP than control group and injury group lens were not enhanced.
Conclusion lens injury can promote the survival of retinal ganglion cells and enhance the expression of GAP-43 and GFAP in retina, and promote axonal regeneration.
Objective to investigate the visual field, visual evoked potential, fundus fluorescein angiography and imaging, and the clinical therapeutic effect of optic neuritis.
Methods 100 cases of optic neuritis patients underwent vision, visual evoked potential, fundus fluorescein angiography and imaging examination, and glucocorticoid, B vitamins, neurotrophic drugs, vasodilators and compound anisodine combined therapy, observe the inspection findings and clinical therapeutic effect.
Results most patients with optic neuritis have no obvious inducement, vision, visual evoked potential and fundus fluorescein angiography have corresponding nonspecific abnormalities, especially magnetic resonance imaging examination is helpful to find abnormal lesions occult, has important clinical value. After comprehensive treatment, visual acuity was improved in 85 cases, 15 cases of visual acuity constant or decreased.
Conclusion it is the key to improve the curative effect and prognosis of patients with optic neuritis to diagnose and take comprehensive treatment in time.

【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R774.61

【参考文献】