Rhodopsin kinase在GK大鼠的视网膜病变中的表达变化和发生机制的研究

发布时间：2018-03-11 02:31

本文选题：糖尿病视网膜病变　切入点：眼底荧光素血管造影　出处：《昆明医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：观察Rhodopsin kinase在不同病程GK大鼠视网膜组织上的表达变化及表达定位,探讨其在糖尿病视网膜病变过程中可能的作用机制,并为进一步深入研究糖尿病视网膜病变的发病机制提供实验依据。方法：选取实验组GK大鼠32只和对照组正常大鼠32只,按糖尿病病程分为四组,即糖尿病4w组、8w组、16w组、24w组,每组8只。对照组与实验组周龄相对应分组,每组8只。采用眼底荧光素血管造影(fundus fluorescein angiography, FFA)在正常大鼠及不同病程糖尿病大鼠模型活体上观察视网膜的变化。采用实时荧光定量多聚酶链反应(Real time-PCR)技术,分析Rhodopsin kinase因子在不同病程糖尿病大鼠视网膜组织上的表达量,并应用免疫组化技术检测Rhodopsin kinase因子在糖尿病大鼠视网膜组织上的表达定位。结果： 1,FFA检查结果显示：总发病率约为82.6%。GK大鼠4周龄即可发现眼底改变,8周以上病程发生眼底改变的几率大大增加。 2, Real time-PCR结果显示：Rhodopsin kinase在对照组大鼠各期的表达变化无明显差异(P0.05)。与对照组相比,糖尿病组各期其表达是下降的,且随着周龄的增加,表达越来越低,即4w时开始降低,24w时降到最低(P0.01)。 3,免疫组化结果显示：Rhodopsin kinase的阳性表达在大鼠视网膜视杆视锥细胞层,外核层,外丛状层;对照组大鼠各期表达无明显差异(P0.05)。糖尿病大鼠4w时,与正常对照组4w相比,阳性强度开始下降,8w与4w相比进一步下降,且随着病程的进展,阳性强度表达越来越低,24w最低(P0.01)。结论： 1,随着病程的增加,DR的发病率逐渐增加,但病程长短并不是DR发展及决定病变的关键因素。不同糖尿病周龄大鼠的血糖值与DR程度也不完全相符. 2, Rhodopsin kinase基因表达的变化可能参与了糖尿病大鼠模型神经视网膜病变的发生发展。神经视网膜病变在2型糖尿病初期即可发生。 3, Rhodopsin kinase基因异常表达引起的光电传导障碍可能是糖尿病视网膜病变早期发生视功能障碍的原因之一。
[Abstract]:Objective:. To observe the expression and localization of Rhodopsin kinase in the retina of rats with different course of disease, and to explore the possible mechanism of the expression of Rhodopsin kinase in the process of diabetic retinopathy. It also provides experimental basis for further study of the pathogenesis of diabetic retinopathy. Methods:. Thirty-two GK rats in the experimental group and 32 normal rats in the control group were selected and divided into four groups according to the course of diabetes, that is, the diabetic group (n = 8), the control group (n = 8) and the control group (n = 8), the control group and the experimental group (n = 8) were divided into four groups according to the course of diabetes. Retinal changes were observed in normal rats and diabetic rats with different course of disease by fundus fluorescein angiography (FFAA) and real-time fluorescence quantitative polymerase chain reaction (real-time fluorescence quantitative polymerase chain reaction), and real-time fluorescence quantitative polymerase chain reaction (Real time-PCR) technique was used to observe the retinal changes in 8 rats in each group. The expression of Rhodopsin kinase factor in the retina of diabetic rats with different course was analyzed. The expression of Rhodopsin kinase factor in the retina of diabetic rats was detected by immunohistochemical technique. Results:. The results of FFA showed that the total incidence rate was about 82.6.GK rats could find the fundus change more than 8 weeks old and the probability of fundus change was greatly increased. 2. The results of Real time-PCR showed that there was no significant difference in the expression of Real Rhodopsin kinase in all stages of the control group. Compared with the control group, the expression of Real time-PCR in each stage of diabetes decreased, and with the increase of age, the expression was lower and lower. In other words, at 4 w, it began to decrease to a minimum of P0.01U at 24 w. 3. The results of immunohistochemistry showed that the positive expression of kinase in the cone cell layer, the outer nuclear layer and the outer plexus layer of the rat retina was not significantly different from that in the control group (P 0.05). When the diabetic rats were 4 weeks old, there was no significant difference between the control group and the normal control group for 4 weeks. The positive intensity began to decrease at 8w and decreased further than that at 4w, and with the progression of disease course, the expression of positive intensity was lower and lower than that of P0.01at 24w. Conclusion:. 1. With the increase of the course of disease, the incidence of Dr increased gradually, but the duration of the disease was not the key factor to determine the development and pathological changes of Dr, and the blood glucose values of the rats of different weeks of diabetes were not completely consistent with the degree of Dr. 2. The change of Rhodopsin kinase gene expression may be involved in the development of neuroretinopathy in diabetic rats, which can occur in the early stage of type 2 diabetes mellitus. 3. The optoelectronic conduction disorder caused by abnormal expression of Rhodopsin kinase gene may be one of the causes of visual dysfunction in the early stage of diabetic retinopathy.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R774.1

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