准分子激光角膜上皮瓣下磨镶术后转化生长因子β1、碱性成纤维细胞生长因子的表达及20%乙醇对兔角膜愈合的影响  

The expression of TGF-β1 and bFGF after LASEK and the effects of 20% ethanol exposure on corneal wound healing

作　　者：王雁[1] 赵堪兴[1] 宿蕾艳[1] 王乐今[2] 耿维莉[1] 金颖[1] 

机构地区：[1]天津市眼科医院天津市眼科研究所,300020 [2]北京大学眼科中心

出　　处：《中华眼科杂志》2005年第6期498-504,共7页Chinese Journal of Ophthalmology

基　　金：天津市自然科学基金资助项目(003611211)

摘　　要：目的探讨准分子激光角膜上皮瓣下磨镶术(LASEK)中乙醇对角膜伤口愈合过程的影响。方法24只健康白兔(48只眼)分为两组,一只眼行机械去上皮法,另一只眼应用20%乙醇30s去除角膜上皮,行相同方案的准分子激光切削,同时设立正常对照组。观察角膜基质浅层角膜细胞数量和形态变化;RTPCR法及免疫组化法检测正常对照组及手术组术后1d、1周、1个月、3个月转化生长因子β1(TGFβ1)和碱性成纤维细胞生长因子(bFGF)的表达情况并做分析。结果术后1d,乙醇组和机械组兔角膜TGFβ1和bFGFRTPCR表达较正常对照组下降;1周两组较正常对照组升高(P<0.01),乙醇组高于机械组(P<0.01);术后1个月达高峰(P<0.01),但乙醇组与机械组比较差异无统计学意义(P>0.05);术后3个月,TGFβ1、bFGFRTPCR产物明显下降,两组之间比较或与正常对照组比较差异无统计学意义(P>0.05)。浅层基质内角膜细胞数量乙醇组与TGFβ1和bFGF的相关系数分别为0.744,0.738;机械组分别为0.664和0.785。结论TGFβ1、bFGF在伤口愈合过程中的表达呈现“减少增多正常”动态过程;20%乙醇在LASEK早期对角膜伤口愈合存在一定影响,但极其短暂。手术中应用是相对安全的。Objective To assess the effects of 20% ethanol used in LASEK on corneal wound healing. Methods Forty-eight eyes from 24 rabbits were deepithelialized by two techniques. The epithelium were detached with either 20% ethanol (applied for 30 seconds) or mechanical scraping, then ablated was performed. The number of superficial stromal keratocytes was counted and the morphologic changes were observed. Expression of TGF-β1 and bFGF mRNAs was detected and analyzed by reverse transcription-polymerase chain reaction and immunocytochemical methods at 1, 7, 30 and 90 days after the surgery. Results One day after the surgery, the expression of TGF-β1 and bFGF in the keratocytes in both treated groups was lower than that of the normal controls. Seven days after the surgery, the expression of TGF-β1 and bFGF in both treated groups was greater than that of the normal controls (P<0.01). The expression in the alcohol-treated group was greater than that of the surgical-treated group (P<0.01). The expression of TGF-β1 and bFGF reached the peak 30 days after the surgery, no significant difference was detected between the alcohol-treated and surgical-treated groups. There was no significant difference in expression level between the alcohol-treated and surgical-treated groups 3 months after the surgery; as well as between treated groups and normal group. The amount of TGF-β1 and bFGF mRNA was positively correlated with the number of keratocytes. The correlation coefficient between TGF-β1 and bFGF mRNA and the number of the keratocytes was 0.744 (P<0.01) and 0.738 (P<0.01) in the alcohol-treated group; and was 0.664 (P<0.01) and 0.785 (P<0.01) in the surgical-treated group, respectively. Conclusions The expression of TGF-β1 and bFGF mRNA undergo a dynamic process of “decrease-increase-normal”. Although ethanol has a slight toxic effect on rabbit epithelial cells, but the effects do not persist over time, therefore, it is relatively safe to use the alcohol treatment in the LASEK.

关 键 词：碱性成纤维细胞生长因子(bFGF) 转化生长因子β1(TGF-β1) 乙醇 准分子激光角膜上皮瓣下磨镶术 术后 角膜愈合 表达及 正常对照组 伤口愈合过程 免疫组化法检测 RT-PCR法 角膜伤口愈合 细胞数量 PCR产物 LASEK 手术中应用 

分 类 号：R779.63[医药卫生—眼科]