本文關(guān)鍵詞： 糖尿病視網(wǎng)膜病變 基質(zhì)金屬蛋白酶2 基質(zhì)金屬蛋白酶9 大鼠　出處：《鄭州大學》2011年碩士論文　論文類型：學位論文

【摘要】：糖尿病視網(wǎng)膜病變(diabetic retinopathy, DR)是糖尿病患者的嚴重并發(fā)癥之一,是導致失明的重要原因,但DR的發(fā)病機制仍在探討中。DR的嚴重程度與糖尿病病程呈正相關(guān)。在對發(fā)病機制的研究中,視網(wǎng)膜毛細血管周細胞損害,破壞了視網(wǎng)膜的內(nèi)屏障,隨后產(chǎn)生了各種血管生成因子和細胞外基質(zhì),促成了新生血管的形成。而增生期糖尿病視網(wǎng)膜病變(proliferative diabetic retinopathy,PDR)的發(fā)生與新生血管的形成有關(guān)。近年來,國內(nèi)外的大量研究針對于PDR的發(fā)病機制,但主要都集中在對血管生成因子的研究上面,而在對細胞外基質(zhì)(extra cellularmatrix, ECM)在PDR的發(fā)病中所發(fā)揮的作用研究甚少。相關(guān)的研究表明,基質(zhì)金屬蛋白酶(matrixmetalloproteinases, MMPs)中MMP-2、MMP-9與新生血管的形成密切相關(guān)。 本實驗擬建立糖尿病眼動物模型,應用免疫組化法測定視網(wǎng)膜中MMP-2、MMP-9蛋白含量的表達,初步探討糖尿病病程的發(fā)展對MMP-2、MMP-9蛋白表達的影響。 材料和方法 選6周齡、健康、清潔級SD大鼠60只,雌雄不限,體重70-90g,常規(guī)母乳喂養(yǎng),排除先天性近視及其它眼疾,隨機分為4組。對照組(Ⅰ組)15只,腹腔注射等量空白緩沖液,飼養(yǎng)21周,其余3組均擬建糖尿病動物模型組,每組15只共45只,禁食水12小時后,按65mg/kg劑量連續(xù)3周左下腹腔小劑量多次注射鏈脲佐菌素(streptozocin,STZ),制作糖尿病大鼠模型。3天后測空腹血糖,血糖"g=16.5mmol/L以上,尿量及飲水明顯增多者為糖尿病大鼠。其中Ⅱ組飼養(yǎng)13周,Ⅲ組飼養(yǎng)17周,Ⅳ組飼養(yǎng)21周,實驗結(jié)束后,分別處死Ⅱ、Ⅲ、Ⅳ及Ⅰ組大鼠,取眼球行視網(wǎng)膜切片HE染色、免疫組織化學染色,通過與電腦相連的Biosens Digital Imaging system V1.6高清晰度彩色病理圖文分析系統(tǒng)作細胞MMP-2蛋白、MMP-9蛋白表達的半定量分析,用平均光密度值來表示免疫反應表達程度。 結(jié)果 1.體重及空腹血糖變化比較：Ⅰ組體重明顯高于Ⅱ組、Ⅲ組及Ⅳ組,其差異具有統(tǒng)計學意義(P0.05)；Ⅱ組、Ⅲ組及Ⅳ組體重兩兩相比無統(tǒng)計學意義。Ⅱ組、Ⅲ組及Ⅳ組空腹血糖明顯高于Ⅰ組,其差異具有統(tǒng)計學意義(P0.05)；Ⅱ組、Ⅲ組與Ⅳ組空腹血糖兩兩相比無統(tǒng)計學意義。 2.免疫組化染色：MMP-2、MMP-9陽性細胞表現(xiàn)為胞漿著色呈棕黃色,主要在神經(jīng)節(jié)細胞層胞漿和血管內(nèi)皮細胞有陽性表達,糖尿病組大鼠隨著病情的發(fā)展,視網(wǎng)膜內(nèi)核層細胞有陽性表達。Ⅱ組、Ⅲ組及Ⅳ組陽性表達均高于Ⅰ組,差異有統(tǒng)計學意義(P0.05)；Ⅱ組、Ⅲ組及Ⅳ組之間比較,差異均有統(tǒng)計學意義(P0.05)。 結(jié)論 1.STZ小劑量多次腹腔注射是一種可靠的制備糖尿病眼動物模型的方法。 2.MMP-2、MMP-9在糖尿病視網(wǎng)膜上表達呈陽性,并隨著糖尿病病程的發(fā)展,二者的表達呈遞增狀態(tài)。
[Abstract]:Diabetic retinopathy (DRR) is one of the serious complications of diabetic patients and an important cause of blindness. However, the pathogenesis of Dr is still under discussion. The severity of Dr is positively correlated with the course of diabetes. In the study of pathogenesis, retinal capillary pericytes damage and damage to the internal barrier of the retina. Various angiogenic factors and extracellular matrix were subsequently produced. And proliferative diabetic retinopathy in proliferative diabetic retinopathy. In recent years, a large number of studies at home and abroad focused on the pathogenesis of PDR, but mainly focused on the study of angiogenic factors. However, the role of extra cellular matrix (ECM) in the pathogenesis of PDR is very little. MMP-9 in matrix metalloproteinases (MMPs) is closely related to angiogenesis. In order to establish an animal model of diabetic eyes, the expression of MMP-2 and MMP-9 in retina was determined by immunohistochemical method, and the relationship between the development of diabetes mellitus and MMP-2 was preliminarily discussed. Effect of MMP-9 protein expression. Materials and methods Sixty healthy, clean SD rats of 6 weeks of age, male and female, weight 70-90 g, were selected. Routine breast-feeding was performed to rule out congenital myopia and other eye diseases. The rats were randomly divided into 4 groups: control group (group 鈪，

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