基质金属蛋白酶-3、9在穿通伤白内障大鼠晶状体上皮细胞的表达及意义

发布时间：2018-08-01 14:14

【摘要】：目的：探讨基质金属蛋白酶-3(matrix metalloproteinase-3, MMP-3)和基质金属蛋白酶-9(matrix metalloproteinase-9, MMP-9)在眼球穿通伤所致的外伤性白内障大鼠晶状体上皮细胞中的表达及意义,探寻MMP-3与MMP-9在穿通伤白内障发病中的作用。方法：健康成年SD大鼠32只,雌雄各半,体质量250-300g,建立单眼晶状体穿通伤后外伤性白内障模型,并随机分为伤后1d、3d、7d、14d 4个时相组。实验开始后在各时间点用裂隙灯显微镜观察大鼠晶状体变化。于伤后1d、3d、7d、14d分别处死各组大鼠,取晶状体,采用免疫组织化学链霉菌亲生物素蛋白-过氧化物酶标(streptavidin-perosidase, SP)二步法检测MMP-3和MMP-9在各组大鼠晶状体上皮细胞(lens epithelial cells, LECs)中的表达情况,扫描电子显微镜(scanning eletron microscope, SEM)观察正常晶状体和穿通伤白内障LECs及晶状体纤维的超微结构。结果： 1.各时相组大鼠正常眼晶状体始终保持透明；32只外伤眼中最终有29眼形成白内障,伤后1d可见晶状体在针头刺穿部位及周围呈淡白色云雾状片状混浊；3d混浊加深、范围扩大；7d混浊范围更大,晶状体大部分呈白色乳块状：14d后其晶状体完全混浊,混浊的晶状体体积明显变小。 2.SEM观察：大鼠正常眼LECs呈不规则扁立方形,整齐单层排列,相邻细胞间连接清晰；晶状体纤维保持完整,定向平行排列,相邻细胞间见指状突彼此镶嵌。大鼠外伤眼LECs游离面呈舌状或片层状,细胞层数增多,相邻细胞间连接增宽；晶状体纤维发生变性,变性纤维失去正常形态,排列紊乱。 3.各时相组大鼠正常眼LECs中MMP-3和MMP-9均无阳性表达,各时相组外伤眼LECs中MMP-3和MMP-9均有阳性表达,与正常眼相比差异有统计学意义(P0.01)。 4.随病程延长,外伤眼LECs中MMP-3和MMP-9阳性表达均逐渐增高,除伤后7d和伤后14d表达差异无统计学意义(P0.05)外,其余各时段间两两比较差异均有统计学意义(P0.05)。 5.MMP-3和MMP-9的阳性表达呈正相关(r=0.967,P0.01)。结论：MMP-3和MMP-9在穿通伤白内障LECs中的表达高于正常眼LECs,且二者存在相关性,提示MMP-3和MMP-9及其相互作用参与了穿通伤白内障形成的病理过程。
[Abstract]:Objective: to investigate the expression and significance of matrix metalloproteinase-3 (MMP-3) and matrix metalloproteinase-9 (MMP-9) in lens epithelial cells of traumatic cataract rats induced by penetrating ocular injury, and to explore the role of MMP-3 and MMP-9 in the pathogenesis of penetrating cataract. Methods: 32 healthy adult SD rats, male and female, with body mass of 250-300g, were used to establish a traumatic cataract model after lens penetrating injury. The rats were randomly divided into 4 groups: 1 day, 3 days after injury, 7 days after injury and 14 days after injury. Slit-lamp microscope was used to observe the changes of rat lens at different time points after the beginning of the experiment. The rats in each group were killed on the 1st day, 3rd day, 7d and 14d after injury. The expression of MMP-3 and MMP-9 in (lens epithelial cells, LECs) of lens epithelial cells of each group was detected by streptavidin-perosidase (SP) two-step method. Scanning electron microscopy (scanning eletron microscope, SEM) was used to observe the ultrastructure of LECs and lens fibers in normal lens and perforated cataract. Results: 1. In each phase group, 29 eyes of 32 traumatic eyes of normal eyes remained transparent, and 1 day after injury, the opacification of the lens was deepened and enlarged in the area of needle-piercing and the surrounding cloud-like opacification for 3 days. The range of opacity was larger at 7 days, and the lens was completely opacity and opacity after 14 days. 2.SEM observation showed that the LECs of normal eyes in rats was irregular and flat square and arranged neatly in single layer. The connection between the adjacent cells was clear, the lens fibers remained intact, the orientation was parallel, and the finger-like processes were embedded among the adjacent cells. The free surface of LECs in traumatic eyes of rats was tongue or lamellar, the number of cell layers increased, the adjacent intercellular junctions widened, and the lens fibers denatured, the denatured fibers lost their normal morphology, and the arrangement was disordered. There was no positive expression of MMP-3 and MMP-9 in LECs of normal eyes in each time phase group, and the positive expression of MMP-3 and MMP-9 in LECs of traumatic eyes in each time phase group, which was significantly different from that in normal eyes (P0.01). 4. With the prolongation of the course of disease, the positive expression of MMP-3 and MMP-9 in traumatic eyes increased gradually, except 7 days after injury and 14 days after injury (P0.05). The positive expression of 5.MMP-3 and MMP-9 were positively correlated with each other (P 0.01). Conclusion the expression of MMP 3 and MMP-9 in penetrating cataract LECs is higher than that in normal eyes, and there is a correlation between them. It is suggested that MMP-3, MMP-9 and their interaction may play an important role in the pathogenesis of penetrating cataract.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R776.1

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