LIF（白血病抑制因子）在小鼠慢性高眼压模型中对GAP-43（生长相关蛋白-43）表达影响的实验研究

发布时间：2018-05-26 01:55

本文选题：慢性高眼压模型 + RGCRGCs　；参考：《山西医科大学》2011年硕士论文

【摘要】：目的：在构建小鼠慢性高眼压模型的基础上，观察白血病抑制因子(Leukemiainhibitory factor, LIF)对生长相关蛋白-43 (growth associated protein-43，GAP-43)在小鼠眼视网膜神经节细胞（retinal ganglion cells，RGCs）及其他部位中的表达影响，探讨其对小鼠慢性高眼压模型中视网膜神经节细胞的保护作用，为临床治疗RGCs呈慢性丧失且病因不清的疾病（如青光眼、视网膜色素变性等）提供一种有效的辅助方法。方法：将60只正常无眼疾昆明小鼠(60只眼)随机分为正常对照组（20只小鼠，20只眼）慢性高眼压组(20只小鼠，20只眼)、LIF治疗组（20只小鼠，20只眼）。每组根据治疗后观察时间分为用药前，用药后1天、7天、14天4个小组（每小组5只小鼠，5只眼）。采用烧灼小鼠巩膜表层的房水引流静脉建立小鼠慢性高眼压模型，于模型成功后（眼压稳定升高1周后）向小鼠玻璃体腔内注射0.5ul/g LIF。分别于用药前，用药后1天、7天、14天摘除眼球，，固定、石蜡包埋、切片，进行HE染色及免疫组织化学染色。普通光学显微镜下观察视网膜形态变化，应用医学图像分析系统对进行RGCs量化及GAP-43表达的灰度分析，进行统计学处理。结果： 1.眼压变化：治疗组与慢性高眼压组在用药前及用药后即刻观察小鼠眼压相较差异无显著性（P0.05）而相较于正常对照组增高，治疗组在随后1天、7天、14天比慢性高眼压组眼压降低，但并不明显，较正常对照组眼压仍高，差异有显著性（P0.05）。 2.组织形态观察：慢性高眼压组织学变化早期为视网膜神经纤维层、内丛状层水肿。晚期视网膜内层水肿消退，萎缩、变薄，视网膜神经节细胞数目减少。治疗组相较于慢性高眼压组水肿消退较快，萎缩不明显，视网膜神经节细胞数目减少不明显。 3.RGCs计数结果：治疗组与慢性高眼压组在治疗前及治疗后1天观察RGCs数目相比较差异无显著性（P0.05）而相较于正常对照组数目减少，慢性高眼压组在随后7天、14天比治疗组RGCs数目减少明显，差异有显著性（P0.05）。但治疗组较正常对照组数目仍少，差异有显著性（P0.05）。 4.免疫组织化学观察：GAP-43表达于各组视网膜神经纤维层、RGCs、以及内丛状层上。通过灰度分析比较，慢性高眼压组早期可见GAP-43的表达增加，随后表达逐渐减少，晚期接近正常；而治疗组早期即可见GAP-43的表达增加，且在观察期内稳定增加，且各期间相较差异有显著性（P0.05）。结论： 1.病理学及眼压改变显示小鼠慢性高眼压模型建模成功。 2.LIF在小鼠慢性高眼压模型中促进GAP-43的表达增加。 3.LIF对小鼠慢性高眼压模型中视网膜神经节细胞具有保护作用。
[Abstract]:Objective: to investigate the effect of Leukemiae inhibitor factor (LIF-43 associated protein-43) on the expression of associated protein 43 in the retinal ganglion cells (RGCs) and other parts of the retinal ganglion cells in mice, and to investigate the effect of Leukemiae inhibitor factor (LIF-43) on the expression of RGCsin the retinal ganglion cells. To investigate its protective effect on retinal ganglion cells (RGCs) in mice with chronic intraocular hypertension, and to provide an effective method for the treatment of RGCs with chronic loss and unclear etiology (such as glaucoma, retinitis pigmentosa, etc.). Methods: sixty eyes of Kunming mice were randomly divided into control group (n = 20, n = 20) and chronic intraocular hypertension group (n = 20, n = 20). Each group was divided into 4 groups according to the time of observation after treatment and 4 groups (5 mice in each group) and 4 groups (5 eyes in each group). A chronic intraocular pressure model was established by cauterizing the aqueous humor drainage veins in the scleral surface of mice. 0.5ul/g lif was injected into the vitreous cavity of mice after the success of the model (1 week after the stable elevation of intraocular pressure). The eyeballs were removed, fixed, embedded in paraffin wax, sectioned, stained with HE and immunohistochemical staining before and 1 day after treatment. The morphological changes of retina were observed under general optical microscope. The quantitative analysis of RGCs and the gray level of GAP-43 expression were analyzed by medical image analysis system. Results: 1. IOP changes: there was no significant difference in intraocular pressure between the treatment group and the chronic high intraocular pressure group before and immediately after treatment (P 0.05), but the IOP in the treatment group was higher than that in the normal control group. The IOP in the treatment group was lower than that in the chronic high intraocular pressure group on the following 7 days and 14 days. However, the IOP was still higher than that in the normal control group, and the difference was significant (P 0.05). 2. Histological changes of chronic intraocular hypertension were retinal nerve fiber layer and edema of internal plexiform layer. Late retinal inner layer edema subsided, atrophy, thinning, retinal ganglion cells decreased. Compared with chronic high intraocular pressure group, edema subsided more quickly, atrophy was not obvious and the number of retinal ganglion cells decreased significantly in the treatment group than in the chronic high intraocular pressure group. Results of 3.RGCs count: there was no significant difference in the number of RGCs between the treatment group and the chronic high intraocular pressure group before treatment and 1 day after treatment (P 0.05), but the number of RGCs in the treatment group was lower than that in the normal control group. The number of RGCs in the chronic intraocular hypertension group was significantly lower than that in the treatment group on the 7th and 14th day after the treatment, and the difference was significant (P 0.05). But the number of the treatment group was still less than that of the normal control group, and the difference was significant (P 0.05). 4. Immunohistochemical observation showed that GAP-43 was expressed in RGCsof retinal nerve fiber layer and on the inner plexiform layer. The expression of GAP-43 in chronic intraocular pressure group was increased at the early stage, then decreased gradually, and then decreased gradually, but the expression of GAP-43 was increased in the treatment group at the early stage, and increased steadily during the observation period. There was a significant difference between different periods (P 0.05). Conclusion: 1. Pathological and intraocular pressure changes showed that the model of chronic high intraocular pressure was successfully established in mice. 2.LIF increased the expression of GAP-43 in mice with chronic intraocular hypertension. 3.LIF can protect retinal ganglion cells from chronic intraocular hypertension in mice.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R775

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