降眼压药物在微球诱导的小鼠高眼压模型中的作用比较

发布时间：2018-05-01 15:37

本文选题：青光眼 + 微球　；参考：《复旦大学》2012年博士论文

【摘要】：研究目的：随着微球诱导的小鼠高眼压模型在眼科基础研究中被广泛地接受和采用,本研究利用该模型检测各种目前临床上常用的青光眼药物的降眼压和视神经保护作用,用于进一步探究该模型在未来青光眼新药筛选中的应用潜力,并尝试使用频域光学相干断层扫描无创性地测量小鼠高眼压模型中的视网膜厚度改变。材料和方法：在氯胺酮(120mg/kg)和甲苯胺噻嗪(12mg/kg)腹腔麻醉下,采用单侧前房注射2微升聚苯乙烯微球悬浮液(5.0×106微球/毫升)的方法在成年C57BL/6J小鼠上建立高眼压模型。比较常用的几种青光眼降眼压药物(0.5%马来酸噻吗洛尔滴眼液、0.15%酒石酸溴莫尼定滴眼液、1%布林佐胺滴眼液、2%盐酸毛果芸香碱滴眼液和0.005%拉坦前列素滴眼液)在这个高眼压模型中的作用。使用TonoLab眼压计监测眼压变化来评价药物的短期和长期降眼压作用。通过对视网膜神经节细胞和视神经轴突计数的方法对视神经损害进行定量分析,比较青光眼药物的视神经保护作用。同时采用频域光学相干断层扫描技术配合自主开发的图像分析软件测量小鼠视网膜神经节细胞复合体的厚度(包括视网膜神经纤维层、视网膜神经节细胞层以及内丛状层),利用该厚度变化比较这青光眼药物的治疗作用。同时分析视网膜神经节细胞复合体厚度变薄率和视网膜神经节细胞丢失率之间的关系。P0.05被认为具有统计学意义。结果：微球注入前房后散布在前房水中,随后逐渐堆积到房角附近,导致了房水外流通道阻塞。眼压随之逐渐升高,在第8天时达到高峰,并且高眼压一直维持到第3周或更长时间。微球注射眼的视网膜神经节细胞和轴突的数量较对侧眼明显减少。我们通过单侧前房注射聚苯乙烯微球的方法成功地建立了小鼠高眼压模型。以抑制房水生成为主要作用机制的降眼压药物(0.5%马来酸噻吗洛尔滴眼液、0.15%酒石酸溴莫尼定滴眼液和1%布林佐胺滴眼液)可以有效降低微球导致高眼压模型鼠的眼压,与对照组相比视网膜神经节细胞和视神经轴突的存活率有明显改善。同时,频域光学相干断层扫描配合自主开发的图像分析软件可以成功地测量小鼠视网膜神经节细胞复合体的厚度。测量结果发现,与对照组相比,以抑制房水生成为主要作用机制的三种降眼压药物均能明显降低由高眼压导致视网膜神经节细胞复合体厚度变薄的幅度。然而以促进房水外流为主要作用机制的降眼压药物(0.005%拉坦前列素滴眼液和2%盐酸毛果芸香碱滴眼液)对微球导致高眼压模型鼠的眼压则完全没有影响。另外,视网膜神经节细胞复合体厚度变薄率与视网膜神经节细胞丢失率之间有明显的相关性。结论：前房注射微球的方法可以有效地诱发小鼠眼压升高并且发生青光眼性的视神经损害。三种房水生成抑制剂在微球导致小鼠高眼压模型中均发挥了有效地降眼压和视神经保护作用。该小鼠高眼压模型是一种房水外流障碍的模型,可有效用于以抑制房水生成为作用机制的降眼压新药的筛选。同时,本研究发现使用光学相干断层扫描测量视网膜神经节细胞复合体厚度为检测青光眼模型小鼠视神经损害、评价药物疗效提供了一个新的生物学指标。
[Abstract]:Objective: with the microsphere induced mouse ocular hypertension model is widely accepted and adopted in the basic research of Ophthalmology, this study uses this model to detect the effect of intraocular pressure and optic nerve protection on various current commonly used glaucoma drugs, and to further explore the potential application potential of the model in the screening of new glaucoma drugs in the future. And try to use frequency domain optical coherence tomography to measure retinal thickness in a mouse model of high intraocular pressure.
Materials and methods: a high intraocular pressure model was established in adult C57BL/6J mice by intraperitoneal anesthesia with ketamine (120mg/kg) and toluidine thiazide (12mg/kg), using a unilateral anterior chamber injection of 2 microsphere polystyrene microspheres (5 x 106 microspheres / milliliters). Several common glaucoma lowering drugs (0.5% maleic acid timolol eye drops) were compared. The role of liquid, 0.15% Brimonidine Tartrate Opthealmic Solution, 1% Brinzolamide Eye Drops, 2% pilocarpine eye drops and 0.005% Latanoprost Eye Drops in this high intraocular pressure model. Using the TonoLab tonometer to monitor intraocular pressure changes to evaluate the short-term and long-term depressurized action of the drug. Through the retinal ganglion cells and the optic deity The method of counting the optic nerve was used to analyze the optic nerve damage and to compare the protective effect of the optic nerve in the glaucoma. The thickness of the retinal ganglion cell complex (including the retinal nerve fiber layer, the retinal ganglion) was measured by the frequency domain optical coherence tomography and the self developed image analysis software. The cell layer and the inner plexiform layer were used to compare the therapeutic effect of this glaucoma drug. The relationship between the thickness thinning rate of the retinal ganglion cell complex and the retinal ganglion cell loss rate (.P0.05) was considered statistically significant.
Results: the microspheres were scattered in the anterior chamber water after the injection of the anterior chamber, and then gradually accumulated near the corner of the room, which resulted in the obstruction of the aqueous Exodus channel. The intraocular pressure increased gradually and reached the peak at eighth days, and the high intraocular pressure remained to third weeks or longer. The number of retinal ganglion cells and axons in the microsphere injection eye was more than the opposite eye. We successfully established a model of high intraocular pressure in mice by injecting polystyrene microspheres into one side of the anterior chamber. In order to inhibit the main mechanism of the aqueous humor of the aqueous humor (0.5% Timolol Maleate Eye Drops, 0.15% Brimonidine Tartrate Opthealmic Solution and 1% Brinzolamide Eye Drops), we can effectively reduce the height of the microspheres. The intraocular pressure of the intraocular pressure model rats was significantly improved compared with the control group. Meanwhile, the frequency domain optical coherence tomography combined with the self developed image analysis software could successfully measure the thickness of the retinal ganglion cell complex in mice. Three kinds of intraocular pressure lowering drugs can significantly reduce the thickness of the retinal ganglion cell complex caused by high intraocular pressure. However, the anti ophthalmic drugs (0.005% Latanoprost Eye Drops and 2% pilocarpine eye drops), which are the main mechanism of promoting aqueous Exodus (0.005% and 2% hairy rutin eye drops), In addition, there is a significant correlation between the thickness thinning rate of the retinal ganglion cell complex and the loss rate of retinal ganglion cells.
Conclusion: the injection of microspheres in the anterior chamber can effectively induce the increase of intraocular pressure in mice and the occurrence of glaucomatous optic nerve damage. Three kinds of aqueous humor inhibitors play an effective role in reducing intraocular pressure and optic nerve protection in the model of high intraocular pressure in mice. The model of high ocular pressure in mice is a model of aqueous humor disorder. At the same time, the use of optical coherence tomography (optical coherence tomography) to measure retinal ganglion cell complex thickness can be used to detect optic nerve damage in glaucoma model mice and to provide a new biological index for evaluating the efficacy of the drug.

【学位授予单位】：复旦大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R775.9

【参考文献】