TIMP-2基因转染FDM豚鼠模型早期后部巩膜MMP-2动态表达研究

发布时间：2018-03-18 00:32

本文选题：形觉剥夺性近视　切入点：巩膜　出处：《青岛大学》2010年硕士论文　论文类型：学位论文

【摘要】： 目的：通过对形觉剥夺性近视豚鼠模型脉络膜上腔注射外源性TIMP-2基因,观察近视早期后极部巩膜MMP-2的表达,探讨外源性TIMP-2基因对后极部巩膜MMP-2表达的影响。方法：采用单眼形觉遮盖法对3周龄三色豚鼠进行形觉剥夺,2w后通过检影验光和测量眼轴长度证实FDM模型是否建立成功。从模型建立成功的豚鼠中随机选取60只,随机分成4组,分别为TIMP-2组(脉络膜上腔注射外源性TIMP-2基因)、空质粒组(脉络膜上腔注射空质粒)、生理盐水组(脉络膜上腔注射生理盐水)、对照组,每组15只。TIMP-2组、空质粒组、生理盐水组对右眼完成注射操作后继续遮盖,其中TIMP-2组左眼为自身对照组；对照组右眼持续遮盖不作任何处理。分别于注射后2d、7d、14d采集后极部巩膜标本,用RT-PCR及Western Blot法检测MMP-2 mRNA及蛋白的表达,统计学分析各组之间及不同时间表达的差异。结果：(1)注射后TIMP-2组MMP-2 mRNA及蛋白的表达出现先降低后增高的变化,注射后2d与7d MMP-2 mRNA及蛋白表达变化有显著性差异(P0.05),7d与14d表达差异无统计学意义(P0.05)。注射后2d、7d、14d,TIMP-2组、自身对照组、对照组之间比较,MMP-2 mRNA及蛋白的表达变化均有统计学意义(均P0.05)；空质粒组、生理盐水组、对照组之间比较,差异无统计学意义(均P0.05)。(2)注射后TIMP-2组TIMP-2 mRNA表达出现先增加后减少的变化。注射后2d与7d表达变化有统计学意义(P0.05),7d与14d差异无统计学意义(P0.05)。注射后2d、7d、14d, TIMP-2组、自身对照组、对照组之间比较,TIMP-2 mRNA表达变化均有统计学意义(均P0.05)；空质粒组、生理盐水组、对照组之间比较,表达变化无统计学意义(均P0.05)。结论：外源性TIMP-2基因转染能明显抑制形觉剥夺性近视眼后极部巩膜MMP-2 mRNA及蛋白的表达,这种改变早期就已经开始发生,随着转染后时间的延长出现先降低后增高的变化。
[Abstract]:Aim: to investigate the effect of exogenous TIMP-2 gene on the expression of MMP-2 in posterior sclera of early myopia by injecting exogenous TIMP-2 gene into the superior choroidal cavity of form-deprived myopia guinea pig model. Methods: the successful establishment of FDM model was confirmed by optometry and eye axis length measurement after 3 weeks old tricolor guinea pigs were deprived of form sensation by monocular shading method. 60 guinea pigs were randomly selected from the successful model. They were randomly divided into four groups: TIMP-2 group (intrachoroidal injection of exogenous TIMP-2 gene), empty plasmid group (suprachoroidal cavity injection of empty plasmids), saline group (intrachoroidal injection of normal saline), control group (15 rats in each group, TIMP-2 group, empty plasmid group). Normal saline group continued to cover the right eye after the injection, in which the left eye of the TIMP-2 group was the self-control group, while the right eye of the control group was continuously covered without any treatment. The sclera specimens were collected at 2 days, 7 days and 14 days after injection, respectively. The expression of MMP-2 mRNA and protein was detected by RT-PCR and Western Blot. Results the expression of MMP-2 mRNA and protein in TIMP-2 group decreased first and then increased after injection. There was no significant difference in the expression of MMP-2 mRNA and protein between 2 and 7 days after injection. There was no significant difference in the expression of MMP-2 mRNA and protein between 7 and 14 days after injection. There were significant differences in the expression of MMP-2 mRNA and protein between the control group and the control group (all P 0.05), the comparison between the blank plasmid group, the normal saline group and the control group. The expression of TIMP-2 mRNA in TIMP-2 group increased first and then decreased after injection. There was no significant difference in the expression of TIMP-2 mRNA between 2 days and 7 days after injection. There was no significant difference in the expression of TIMP-2 mRNA between 2 days and 7 days after injection. There was no significant difference in the expression of TIMP-2 mRNA between 2 days after injection and 14 days after injection (P 0. 05%). The expression of TIMP-2 mRNA in TIMP-2 group was 14 days after injection, while in TIMP-2 group and self control group, there was no significant difference between 2 days and 14 days after injection. The changes of TIMP-2 mRNA expression in the control group were statistically significant (P 0.05), but there was no significant difference between the blank plasmid group, normal saline group and control group (all P 0.05). Conclusion: exogenous TIMP-2 gene transfection can significantly inhibit the expression of MMP-2 mRNA and protein in the posterior sclera of form-deprived myopia, and this change has begun to occur at the early stage. With the extension of transfection time, the expression of MMP-2 mRNA and protein in the posterior sclera is decreased first and then increased.
【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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