TIMP-2基因转染豚鼠FDM模型生物学和巩膜形态学改变

发布时间：2018-05-05 03:15

本文选题：近视 + 形觉剥夺　；参考：《青岛大学》2010年硕士论文

【摘要】： 目的：观察基质金属蛋白酶抑制剂-2(tissue inhibitor of matrixmetalloproteinase-2,TIMP-2)基因转染豚鼠形觉剥夺性近视(form deprivation myopia, FDM)模型屈光度、眼轴变化及后极部巩膜形态学改变,探讨TIMP-2基因对形觉剥夺近视有无作用。方法：4周龄健康豚鼠60只,随机分成TIMP-2组、空质粒组、生理盐水组和对照组四个组,每组15只。右眼为剥夺眼,采用半透明眼罩遮盖诱导轴性近视,左眼不予处理。选取TIMP-2组豚鼠左眼作为自身对照组。TIMP-2组、空质粒组、生理盐水组形觉剥夺后15天于右眼脉络膜上腔分别注射转染TIMP-2基因的脂质体、转染空质粒的脂质体和生理盐水。在注药后2、7、14天进行各组双眼屈光度、眼轴检测。然后处死豚鼠。光镜观察巩膜的厚薄,成纤维细胞的密度及细胞外基质的变化。透射电镜观察胶原纤维的排列、直径大小及细胞外基质的变化。结果：TIMP-2组屈光度在注药后第7天开始有改变,第14天达到高峰。注药后2、7、14天TIMP-2组右眼屈光度分别为-0.4±0.5 D,0.9±0.6D,1.2±0.6D,第14天TIMP-2组与其他三个实验组差别经统计学处理有显著差异(P0.05)；注药后2、7、14天TIMP-2组眼轴分别为7.78±0.04mm,7.69±0.03mm,7.65±0.03mm,其中第7、14天TIMP-2组与其他三个实验组差别经统计学处理有显著差异(P0.05)。光镜观察注药后TIMP-2组后极部巩膜明显变厚,成纤维细胞密度增高,细胞外基质减少。电镜观察发现TIMP-2组剥夺眼巩膜胶原纤维分布逐渐密集,胶原纤维直径有所增大。TIMP-2组注药后2、7、14天胶原纤维平均直径分别为49.7nm,71.3nm,97.8nm,与自身对照组比较差别经统计学处理均有统计学意义(P0.05)。在第2天差别与其他三个实验组相比,在第7、14天差别经统计学处理均有统计学意义(P0.05)。结论：TIMP-2基因注入豚鼠形觉剥夺性近视模型,可引起巩膜变厚、眼轴变短、胶原纤维直径增大,屈光度下降。二者的改变同近视程度减弱的发展趋势一致,且形态学改变先于生物学改变。因此TIMP-2基因对于豚鼠形觉剥夺性近视模型形态学和生物学改变有一定的抑制效应。
[Abstract]:Objective: to investigate the effect of TIMP-2 gene on form-deprivation myopia deprivation myopia, FDM) model in guinea pigs transfected with matrix metalloproteinase-2 inhibitor of matrix metalloproteinase-2 (MMP-2) gene. Methods Sixty healthy guinea pigs at the age of 4 weeks were randomly divided into four groups: TIMP-2 group, empty plasmid group, normal saline group and control group, with 15 in each group. The right eye is deprived, translucent goggles are used to cover induced axial myopia, left eye is not treated. The left eyes of guinea pigs in TIMP-2 group were selected as self control group. TIMP-2 group, blank plasmid group and normal saline group were injected with liposomes transfected with TIMP-2 gene in the suprachoroidal cavity of right eye 15 days after form deprivation, and the empty plasmid liposomes and normal saline were transfected respectively. The diopter and eye axis of each group were measured at 2 minutes and 14 days after injection. Then the guinea pigs were killed. The thickness of sclera, the density of fibroblasts and the changes of extracellular matrix were observed under light microscope. The arrangement, diameter and extracellular matrix of collagen fibers were observed by transmission electron microscope. Results the diopter of the 1-TIMP-2 group began to change on the 7th day after injection and reached the peak on the 14th day. The diopter of right eye in TIMP-2 group was -0.4 卤0.5 DU 0.9 卤0.6Dl 1.2 卤0.6D on day 14 after injection, there was significant difference between TIMP-2 group and other three experimental groups on the 14th day (P 0.05), and the ocular axis in TIMP-2 group was 7.78 卤0.04 mm 7.69 卤0.03 mm on the 14th day after injection, which was 7.65 卤0.03 mm in TIMP-2 group on day 714 and the other three groups respectively, and the diopter was 7.65 卤0.03 mm in TIMP-2 group on day 714 after injection. There was a significant difference between the two groups by statistical analysis (P 0.05). Under light microscope, the sclera became thicker, the density of fibroblasts increased and the extracellular matrix decreased in TIMP-2 group. Electron microscope observation showed that the distribution of collagen fibers in the sclera of TIMP-2 group was gradually dense, and the diameter of collagen fibers was increased. The average diameter of collagen fibers in the TIMP-2 group was 49.7 nm, 71.3 nm and 97.8 nm, respectively. The difference was statistically significant compared with the control group (P 0.05). On the second day, the difference was statistically significant compared with the other three experimental groups, and on the 7th and 14th day, the difference was statistically significant (P 0.05). Conclusion injection of 1: TIMP-2 gene into the shape deprivation myopia model of guinea pigs can result in sclera thickening, eye axis becoming shorter, collagen fiber diameter increasing, and diopter decreasing. The changes were consistent with the weakening of myopia, and the morphological changes were prior to biological changes. Therefore, TIMP-2 gene has a certain inhibitory effect on morphological and biological changes of form-deprived myopia in guinea pigs.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R778.11

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