本文選題：糖尿病視網(wǎng)膜病變　切入點：眼底熒光素血管造影　出處：《昆明醫(yī)科大學》2012年碩士論文　論文類型：學位論文

【摘要】：目的： 觀察Rhodopsin kinase在不同病程GK大鼠視網(wǎng)膜組織上的表達變化及表達定位,探討其在糖尿病視網(wǎng)膜病變過程中可能的作用機制,并為進一步深入研究糖尿病視網(wǎng)膜病變的發(fā)病機制提供實驗依據(jù)。 方法： 選取實驗組GK大鼠32只和對照組正常大鼠32只,按糖尿病病程分為四組,即糖尿病4w組、8w組、16w組、24w組,每組8只。對照組與實驗組周齡相對應(yīng)分組,每組8只。采用眼底熒光素血管造影(fundus fluorescein angiography, FFA)在正常大鼠及不同病程糖尿病大鼠模型活體上觀察視網(wǎng)膜的變化。采用實時熒光定量多聚酶鏈反應(yīng)(Real time-PCR)技術(shù),分析Rhodopsin kinase因子在不同病程糖尿病大鼠視網(wǎng)膜組織上的表達量,并應(yīng)用免疫組化技術(shù)檢測Rhodopsin kinase因子在糖尿病大鼠視網(wǎng)膜組織上的表達定位。 結(jié)果： 1,FFA檢查結(jié)果顯示：總發(fā)病率約為82.6%。GK大鼠4周齡即可發(fā)現(xiàn)眼底改變,8周以上病程發(fā)生眼底改變的幾率大大增加。 2, Real time-PCR結(jié)果顯示：Rhodopsin kinase在對照組大鼠各期的表達變化無明顯差異(P0.05)。與對照組相比,糖尿病組各期其表達是下降的,且隨著周齡的增加,表達越來越低,即4w時開始降低,24w時降到最低(P0.01)。 3,免疫組化結(jié)果顯示：Rhodopsin kinase的陽性表達在大鼠視網(wǎng)膜視桿視錐細胞層,外核層,外叢狀層;對照組大鼠各期表達無明顯差異(P0.05)。糖尿病大鼠4w時,與正常對照組4w相比,陽性強度開始下降,8w與4w相比進一步下降,且隨著病程的進展,陽性強度表達越來越低,24w最低(P0.01)。 結(jié)論： 1,隨著病程的增加,DR的發(fā)病率逐漸增加,但病程長短并不是DR發(fā)展及決定病變的關(guān)鍵因素。不同糖尿病周齡大鼠的血糖值與DR程度也不完全相符. 2, Rhodopsin kinase基因表達的變化可能參與了糖尿病大鼠模型神經(jīng)視網(wǎng)膜病變的發(fā)生發(fā)展。神經(jīng)視網(wǎng)膜病變在2型糖尿病初期即可發(fā)生。 3, Rhodopsin kinase基因異常表達引起的光電傳導(dǎo)障礙可能是糖尿病視網(wǎng)膜病變早期發(fā)生視功能障礙的原因之一。
[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.
【學位授予單位】：昆明醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R774.1

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