TIMP-2基因转染FDM豚鼠模型早期后极部巩膜中MMP-2的动态表达  

Dynamic expression of MMP-2 in posterior pole sclera after exogenous TIMP-2 gene transfection in form deprivation myopia guinea pigs

作　　者：李勇子[1] 刘桂香[1] 王玲[1] 隋爱玲[2] 郭强强[3] 万鲁芹 

机构地区：[1]青岛大学医学院附属医院眼科,山东省青岛市266003 [2]青岛大学医学院附属医院中心实验室,山东省青岛市66003 [3]沧州医学高等专科学校,河北省沧州市61001 [4]青岛眼科医院,山东省青岛市266000

出　　处：《眼科新进展》2012年第12期1104-1109,共6页Recent Advances in Ophthalmology

基　　金：山东省科技厅攻关计划(编号:2007GG30002025)~~

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组织金属蛋白酶抑制剂-2(tissue inhibitor of matrix metalloproteinases,TIMP-2)基因,观察近视早期后极部巩膜中基质金属蛋白酶-2(matrix metalloproteinase-2,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。方法采用单眼遮盖法对3周龄三色豚鼠进行形觉剥夺,2周后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质粒)、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做任何处理。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、TIMP-2 mRNA及MMP蛋白的表达,统计分析各组之间不同时间的表达差异。结果 TIMP-2组注射后2d与7dMMP-2 mRNA及蛋白表达变化差异有统计学意义(均为P<0.05),7d与14d表达差异无统计学意义(均为P>0.05)。注射后2d、7d、14d,TIMP-2组与自身对照组、对照组之间比较,MMP-2 mR-NA及蛋白的表达变化均有统计学意义(均为P<0.05);空质粒组、生理盐水组、对照组之间比较,差异无统计学意义(均为P>0.05)。TIMP-2组注射后2d与7dTIMP-2 mRNA表达变化差异有统计学意义(P<0.05),7d与14d差异无统计学意义(均为P>0.05)。注射后2d、7d、14dTIMP-2组与自身对照组、对照组之间比较,TIMP-2 mRNA表达差异均有统计学意义(均为P<0.05);空质粒组、生理盐水组、对照组之间比较,TIMP-2 mRNA表达差异均无统计学意义(均为P>0.05)。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,随着转染时间的延长出现先降低后增高的变化。Objective To investigate the effects of exogenous tissue inhibitor of matrix metalloproteinase-2 （ TIMP-2 ） gene on matrix metalloproteinase-2 （ MMP-2 ） at posterior pole sclera by observing the expression of MMP-2 in early myopia after exoge- nous TIMP-2 gene injection in form deprivation myopia（FDM） guinea pigs. Methods The 3-week-old tri-color guinea pigs were monocular deprived to establish FDM model. Sixty FDM guinea pigs, which were selected randomly 2 weeks later, were divided into 4 groups randomly： TIMP-2 group （the exogenous TIMP-2 gene was injected into supra- choroidal space）, empty vector group （the empty plasmid was injected into suprachoroi- dal space ）, normal saline group （the normal saline was injected into suprachoroidal space） and control group, 15 cases in each group. The right eyes of the TIMP-2 group, empty vector group and normal saline group were deprived again after injections while the control group without any treatment. The left eyes of TIMP-2 group were set as serf- control group. The posterior pole sclera was collected at 2 days ,7 days and 14 days after injection. The mRNA of MMP and TIMP and protein of MMP-2 were detected by RT-PCR and Western blot, and the results between different groups as well as different time points were analyzed by statistical analysis. Results There was statistical difference in mRNA and protein of MMP-2 between 2 days and 7 days after injection in TIMP-2 group （P 〈 0.05 ）, while no difference between 7 days and 14 days （ P 〉 0.05 ）. The differ- ences were significant in mRNA and protein of MMP-2 between TIMP-2 group,self-con- trol group and control group at 2 days, 7days and 14 days （ all P 〈 0.05 ）. However, the difference was not significant between empty vector group, normal saline group and control group at each time point（ all P 〉 0.05 ）. There was statistical difference in TIMP- 2 mRNA between 2 days and 7 days after injection in TIMP-2 group（P 〈 0.05 ） ,while no difference between 7 days and 1

关 键 词：形觉剥夺性近视 巩膜 基质金属蛋白酶 基质金属蛋白酶组织抑制剂 

分 类 号：R730.59[医药卫生—肿瘤]