表层切削术后早期应用碱性成纤维细胞生长因子对兔角膜上皮下雾状混浊的作用及其机制  

Effects of early applying basic fibroblast growth factor on corneal haze after surface ablation surgery and its mechanism

作　　者：隋文君 高洪莲[2] 刘奇奇 张磊[1] Sui Wenjun;Gao Honglian;Liu Qiqi;Zhang Lei(Department of Ophthalmology,Affiliated Hospital of Binzhou Medical University,Binzhou 256600,China;Clinical Medicine Laboratory,Affiliated Hospital of Binzhou Medical University,Binzhou 256600,China)

机构地区：[1]滨州医学院附属医院眼科,山东省滨州市256600 [2]滨州医学院附属医院临床医学实验室,山东省滨州市256600

出　　处：《中华实验眼科杂志》2020年第11期936-943,共8页Chinese Journal Of Experimental Ophthalmology

基　　金：山东省医药卫生科技发展计划项目(2016WS0022)。

摘　　要：目的探讨兔角膜表层切削术后早期应用碱性成纤维细胞生长因子(bFGF)对角膜上皮下雾状混浊(haze)的作用及其机制。方法选取60只健康普通级新西兰大白兔,均行右眼准分子激光角膜切削术(PRK),术后采用随机数字表法将实验眼分为PRK+生理盐水组、PRK+bFGF组和单纯PRK组,每组20眼,术后分别给予生理盐水和bFGF点眼,每日3次,每次1滴,单纯PRK组未行任何处理,药物连续应用至动物处死,另取8只兔为空白对照组。各手术组分别于术后7d和28d各取10只兔,空白对照组取4只兔,通过眼前节照相、眼前节光相干断层扫描成像(AS-OCT)记录角膜上皮愈合及haze情况,并行Fantes分级;采用苏木素-伊红染色法观察兔角膜组织病理学变化;采用免疫组织化学法检测兔角膜组织中转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)、基质金属蛋白酶-2(MMP-2)的表达。结果各手术组角膜上皮愈合时间比较差异无统计学意义(F=0.57,P=0.57)。各组兔眼术后不同时间点haze分级总体比较差异均有统计学意义(F组别=41.736,P<0.01;F时间=129.445,P<0.01);术后28d,PRK+bFGF组haze分级明显高于PRK+生理盐水组和单纯PRK组,差异均有统计学意义(均P<0.05)。AS-OCT显示,空白对照组角膜上皮表面反射条带连续、光滑,上皮细胞与基质层排列紧密;术后7d各组角膜上皮表面反射条带连续、光滑,与浅基质层连接欠紧密,浅基质层反射增强,组织病理学检查发现术区角膜上皮细胞和基质层细胞增生,基质层胶原纤维排列紊乱,其中以PRK+bFGF组最明显;术后28d,上述表现明显增强。免疫组织化学染色结果显示,空白对照组角膜组织中仅见少量MMP-2表达;术后7d各手术组角膜组织中TGF-β1、α-SMA、MMP-2均呈阳性表达,PRK+bFGF组角膜组织中阳性染色最强;术后28d,TGF-β1、α-SMA和MMP-2的表达较术后7d明显增加,差异均有统计学意义(均P<0.05)。结论表层切削术后早期Objective To investigate the effects of early applying of basic fibroblast growth factor(bFGF)on corneal haze formation after surface ablation surgery in rabbits.Methods The right eyes of 60 healthy New Zealand white rabbits received photorefractive keratectomy(PRK)and were randomized into PRK+normal saline group,PRK+bFGF group and simple PRK group,with 20 rabbits in each group.Normal saline solution and bFGF were topically administered according to grouping,respectively,3 times per day,1 drop for each time until the sacrifice of the animals,and no drug was used in the PRK group.Another 8 normal rabbits were served as blank control group.The corneal healing response and haze formation were evaluated by anterior segment photography and anterior segment optical coherence tomography(AS-OCT)and graded based on Fantes criteria.Corneal histopathology was examined by hematoxylin-eosin staining.Immunohistochemistry was used to detect the expression of transforming growth factor-β1(TGF-β1),α-smooth muscle actin(α-SMA)and matrix metalloproteinase-2(MMP-2)in cornea.This study protocol was approved by the Experimental Animal Ethic Committee of Affiliated Hospital of Binzhou Medical University(20180209-03).The use and care of the animals complied with the Statement of ARVO.Results The corneal epithelium was completely healed in 3-4 days following surgery and there was not significantly different in healing time among the three groups.(F=0.57,P=0.57).The haze grading was significantly different among different groups at different time points(Fgroup=41.736,P<0.01;F time=129.445,P<0.01)and showed the highest score in the PRK+bFGF group on the 28th day after operation.On the 7th day after surgery,AS-OCT image showed that the surface reflection of corneal epithelium was continuous and smooth and corneal epithelium was not tightly attached to the superficial stromal layer;the reflection of the superficial stromal layer was enhanced in all the operation groups.The proliferation of corneal epithelial cells and superficial stromal

关 键 词：转化生长因子-Β1 Α-平滑肌肌动蛋白 基质金属蛋白酶-2 碱性成纤维细胞生长因子 角膜上皮下雾状混浊 表层切削术 

分 类 号：R779.6[医药卫生—眼科]