豚鼠早期实验性近视眼巩膜成纤维细胞前部及后极部MMP-2 mRNA及蛋白表达变化的研究  

Expression of MMP-2 in anterior and posterior scleral fibroblast in early experimental myopia of guinea pig

作　　者：陈博宇[1] 王超英[1] 安慧琴[2] 马景学[3] 陈维毅[4] 郝岚[1] 刘迎庆[1] 仝春梅[1] 

机构地区：[1]白求恩国际和平医院眼科,河北省石家庄市050082 [2]河北省人民医院耳鼻喉科,河北省石家庄市050000 [3]河北医科大学第二附属医院眼科,河北省石家庄市050000 [4]太原理工大学生物力学研究所,山西省太原市030001

出　　处：《眼科新进展》2011年第9期807-811,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:10872140)~~

摘　　要：目的研究豚鼠早期实验性近视眼巩膜成纤维细胞(guineapig scleral fibroblast,GSF)前部及后极部基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)的改变,探索近视的发病机制。方法 2周龄豚鼠30只作为实验组,另外再随机选取5只(10眼)正常2周龄豚鼠不作任何干预,作为正常对照眼。选取任意眼戴-10.00DS凹透镜,分别饲养6d、15d、30d后除去镜片,验光及测眼轴长度确定近视形成后,采用组织块培养法培养豚鼠的前部及后极部GSF,利用免疫细胞化学、实时荧光定量PCR、Western blot蛋白印迹法检测前部及后极部GSF中MMP-2表达变化。结果 MMP-2的表达定位于细胞浆和细胞核。实验组中前部GSF饲养15d阳性表达率较高,30d阳性表达率最高;后极部GSF饲养6d表达率开始较高,至15d阳性表达率最高,以后保持较高的阳性表达率,各诱导时间点之间两两比较,差异均有统计学意义(均为P<0.05);自身对照组各组自身前部及后极部比较,差异均无统计学意义(均为P>0.05)。结论体外培养的豚鼠GSF稳定表达MMP-2 mRNA及其蛋白,且前部GSF于诱导15d开始阳性表达率较高,后极部GSF于诱导6d开始阳性表达率较高,后极部GSF较前部表达明显增多。Objective To determine the expression of matrix metalloproteinase 2（MMP-2） in anterior and posterior scleral fibroblasts in early experimental myopia of guinea pig,and study the pathogenesis of myopia.Methods Thirty guinea pigs with 2 weeks old were set as the experimental group,meanwhile 5 normal pigs（10 eyes） with 2 weeks old were randomly chosen and set as the control group without any treatment.The optional eye of experimental group wore the concave lens with-10.00 DS,the lens were removed after 6 days,15 days and 30 days,the optometry and ocular axis were examined to determine the myopia formation,the anterior and posterior scleral fibroblasts of each group were cultured and passaged from 3 generations to 6 generations,the expression of MMP-2 in passaged scleral fibroblasts cells was examined by immunocytochemistry,RT-PCR and Western blot.Results The expression of MMP-2 was located in cytoplasm and nucleus.In the experimental group,the positive expression rat in anterior scleral fibroblasts at 15 days was higher,30 days was the most,which in posterior scleral fibroblasts at 6 days was higher,15 days was the most,and maintained later,there were significant difference between each two time points（all P0.05）.There was no statistical difference of MMP-2 expression between anterior and posterior of self-control eyes（all P0.05）.Conclusion The expression of MMP-2 mRNA and protein in scleral fibroblasts cells cultured in vitro of guinea pigs is stable,the positive rates are in high from 15 days in anterior and 6 days in posterior,the expression of MMP-2 in posterior is higher than that in anterior scleral fibroblasts cells.

关 键 词：豚鼠 巩膜成纤维细胞 基质金属蛋白酶 2 免疫细胞化学 实时荧光定量PCR Western blot蛋白印迹法 

分 类 号：R777.2[医药卫生—眼科]