豚鼠早期实验性近视眼视网膜色素上皮细胞bFGF表达变化的研究  

Altered bFGF expression in retinal pigment epithelial cells from guinea pig model with early stage of experimentally-induced myopia

作　　者：陈博宇[1] 王超英[1] 陈维毅[2] 刘迎庆[1] 郝岚[1] 

机构地区：[1]白求恩国际和平医院眼科,中国河北省石家庄市050082 [2]太原理工大学生物力学研究所,中国山西省太原市030001

出　　处：《国际眼科杂志》2015年第11期1857-1861,共5页International Eye Science

基　　金：河北省自然科学基金资助项目(No.H2012505009)~~

摘　　要：目的：研究豚鼠早期实验性近视眼视网膜色素上皮（retinal pigment epithelial,RPE）细胞前部及后极部碱性成纤维细胞生长因子（basic fibroblast growth factor,b FGF）的改变,探索近视的发病机制。方法：两周龄豚鼠30只随机分为A组、B组、C组,每组10只,再随机选取5只10眼正常两周龄豚鼠不作任何干预,作为正常对照眼。选取任意眼戴一-10.00D凹透镜,分别饲养6、15、30d后除去镜片,验光及测眼轴长度确定近视形成后,采用细胞酶消化法培养豚鼠的前部及后极部RPE细胞,取3~6代RPE细胞进行免疫细胞化学、实时荧光定量PCR、Western-blot蛋白印迹法检查前部及后极部RPE细胞中b FGF表达变化。结果：b FGF的表达定位于细胞浆和细胞核。免疫细胞化学、实时荧光定量PCR、Western-blot蛋白印迹法结果均表明：A、B、C三组实验眼和对照眼前部及后极部RPE细胞均有b FGF的表达,实验组前部及后极部与对照组相应前部及后极部比较,实验组中b FGF的阳性率低于对照组,差异有统计学意义（P〈0.05）;而且,随着诱导时间的推移,实验组的阳性率逐渐降低,差异有统计学意义（P〈0.05）,对照组的阳性率不变,差异无统计学意义（P〉0.05）;但实验组和对照组各组自身前部及后极部比较,差异无统计学意义（P〉0.05）。结论：实验组前部及后极部与对照组相应前部及后极部比较,b FGF的表达显著低于对照组。AIM：To determine the expression of basic fibroblast growth factor（bFGF） in retinal pigment epithelial（RPE）cells from anterior and posterior part of guinea pig eyes with lens-induced myopia（LIM）.METHODS：Thirty two-week-aged guinea pigs were randomly divided into three groups,group A,group B and group C.Another five guinea pigs（10 eyes） were chosen as control group without any treatment.Concave lenses were worn by the 30 guinea pigs on either side of their eyes.After 6,15 and 30 d,the lenses were removed and optometry and axial length were used to make sure the myopia had formed.The retinal pigment epithelial cells of each group were cultured and passaged in vitro using enzymatic digestion method.The cultured cells at3~6 generation was detect by immunocytolchemistory,Real-Time PCR and Western blotting to detect the expression of bFGF.RESULTS：The expression of bFGF was located in the cytoplasm and nucleus.The results of immunocytolchemistory,Real-Time PCR and Western blotting showed that the expression of bFGF were detected at anterior and posterior part of eyes from experiment group and control group,the expression in experiment group at both part was significantly lower than those in control group（P〈0.05）.The positive percentage of expression of bFGF in experiment group decreased with the induced time（P〈0.05）.The expression of bFGF from anterior part of both experiment group and control group was as the same as that from posterior part of themselves（P〉0.05）.CONCLUSION：The expression of bFGF from anterior and posterior part of experiment group was lower than those of control group.

关 键 词：豚鼠 视网膜色素上皮细胞 碱性成纤维细胞生长因子 免疫细胞化学 实时荧光定量PCR Western-blot蛋白印迹法 

分 类 号：R778.11[医药卫生—眼科]