大鼠激素性高眼压模型中眼内HIC-5表达的实验研究

发布时间：2018-03-19 11:16

本文选题：激素性青光眼　切入点：大鼠　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:建立大鼠激素性高眼压模型,观察激素性高眼压大鼠模型中眼内HIC-5的表达,初步探讨HIC-5在激素性青光眼发病中的作用。方法:健康的成年雄性SD大鼠90只,随机分为地塞米松组40只、灭菌注射用水组25只、空白对照组25只。地塞米松组大鼠右眼球结膜下隔日定时注射地塞米松磷酸钠注射液0.2m L(1m L:5mg)制作激素性高眼压动物模型,灭菌注射用水组大鼠右眼球结膜下隔日定时注射0.2m L灭菌注射用水,空白对照组大鼠不做任何处理。动物模型制作前使用回弹式眼压计测量大鼠眼压1周,计算大鼠眼压的正常区间,球结膜下注射前、后分别使用回弹式眼压计测量大鼠眼压。造模时间30天,以造模结束时,大鼠眼压较处理前平均值升高10mm Hg以上作为造模成功依据。造模结束后处死各组大鼠,利用免疫荧光组化染色及Western-blot检测各组大鼠房角及其周围组织中HIC-5的表达。结果:地塞米松组共有21只大鼠成功诱发激素性高眼压模型,记为模型组;于灭菌注射用水组及空白对照组各随机选取21只大鼠,分别记为无菌水组、空白对照组。三组大鼠右眼处理前,眼压相比差异无统计学意义,其正常眼压区间为10.65±0.53mm Hg。模型组大鼠右眼经地塞米松磷酸钠溶液处理后第8天眼压开始升高,第8-16天眼压持续升高,24-30天眼压趋于平稳。无菌水组大鼠右眼处理前后相比眼压变化无统计学意义。大鼠眼球冰冻切片房角组织可观察到孔样的小梁网、虹膜睫状体、Schlemm管等结构,HIC-5阳性表达于各组大鼠房角及及其周围组织。与无菌水组、空白对照组大鼠相比,模型组大鼠经地塞米松磷酸钠溶液处理后,HIC-5在房角及其周围组织的荧光强度明显增强,表达量增多,差异具有统计学意义(F=220.06,P0.01)。Western blot检测结果显示与无菌水组、空白对照组相比,模型组大鼠房角及其周围组织中HIC-5的灰度值增高,表达量上升,差异具有统计学意义(F=27.62,P0.01)。结论:大鼠球结膜下注射地塞米松磷酸钠溶液可以成功制作激素性高眼压模型;大鼠激素性高眼压模型中眼内HIC-5含量表达升高,可能与激素性青光眼的发生相关。
[Abstract]:Objective: to establish a steroid-induced intraocular pressure (IOP) model in rats, observe the expression of intraocular HIC-5 in steroid-induced intraocular hypertension (IOP) rats, and explore the role of HIC-5 in the pathogenesis of steroid-induced glaucoma. The rats in the dexamethasone group were randomly divided into dexamethasone group (n = 40), sterilized injection group (n = 25) and blank control group (n = 25). The right eye subconjunctival subconjunctiva of sterilized rats was injected with 0.2m L sterilizing water every other day, while the blank control group did not do any treatment. The intraocular pressure of rats was measured by rebound IOP for 1 week before the establishment of the animal model. The normal interval of intraocular pressure in rats was calculated. The intraocular pressure was measured by rebound intraocular pressure meter before and after subconjunctival injection. The intraocular pressure (IOP) of the rats was increased by more than 10mm Hg compared with the average before treatment as the basis for successful modeling. Immunohistochemical staining and Western-blot were used to detect the expression of HIC-5 in the atrial horn and surrounding tissues of rats. Results: Twenty-one rats in dexamethasone group were successfully induced steroid-induced intraocular hypertension (IOP) model. Twenty-one rats were randomly selected as aseptic water group and blank control group in the sterilization injection water group and the blank control group respectively. There was no significant difference in IOP between the three groups before the right eye treatment. The normal IOP interval was 10.65 卤0.53mm Hg.Intraocular pressure began to increase on the 8th day after the right eye was treated with dexamethasone sodium phosphate (Dexamethasone sodium phosphate solution) in the model group. From the 8th to 16th day, the IOP increased steadily from 24 to 30 days. There was no significant difference in intraocular pressure between the right eye of rats treated with aseptic water before and after treatment. The trabecular meshwork like holes could be observed in the cornea of frozen section of rat eyeball. The positive expression of HIC-5 was found in the angle of the atrium and its surrounding tissues in all groups, compared with the sterile water group and the blank control group. After treated with dexamethasone sodium phosphate solution, the fluorescence intensity and expression of HIC-5 in the horn and surrounding tissues of the model group were significantly increased, and the difference was statistically significant (P 0.01). The results of Western blot analysis showed that compared with the aseptic water group and the blank control group, the expression of HIC-5 in the model group was significantly higher than that in the sterile water group and the blank control group. In the model group, the gray level and expression of HIC-5 in the atrial angle and its surrounding tissues were increased, and the difference was statistically significant. Conclusion: injecting dexamethasone sodium phosphate solution under the conjunctiva of rats can successfully make the steroid-induced intraocular pressure model. The increased expression of HIC-5 in the rat model of high intraocular pressure may be related to the occurrence of steroid-induced glaucoma.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R775;R-332

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