碱性成纤维细胞生长因子和表皮生长因子对角膜中期保存液保存人角膜内皮细胞的影响  

The effect of basic fibroblast growth factor and epidermal growth factor on corneal endothelial cells preserved in mid-term preserving medium

作　　者：闫超[1] 王立[1] 潘志强[1] 

机构地区：[1]首都医科大学附属北京同仁医院北京同仁眼科中心北京同仁眼库,100730

出　　处：《中华眼科杂志》2006年第2期166-170,共5页Chinese Journal of Ophthalmology

摘　　要：目的探讨碱性成纤维细胞生长因子(bFGF)和表皮生长因子(EGF)对中期保存角膜内皮细胞活性的影响。方法角膜中期保存液保存人角膜环,对照组为角膜中期保存液,实验组分为A、B、C、D组,分别为角膜中期保存液中加入bFGF(5ng/ml)、bFGF(20ng/ml)、EGF及bFGF+EGF,在保存角膜第3、7、14天后分别取保存角膜环复温观察,锥虫蓝茜素红联合染色检测角膜内皮活细胞率,电镜检测细胞超微结构的改变。结果保存角膜环第14天角膜内皮活细胞率,对照组为(10.35±1.32)%,A组为(62.18±1.56)%,B组为(92.57±0.90)%,C组为(71.01±2.67)%,D组(82.59±1.45)%,与对照组比较,各保存时间各实验组活细胞率均高,差异有统计学意义(P<0.01)。保存第14天,对照组保存的角膜环明显水肿、混浊不透明、内皮细胞形态结构不完整,超微结构显示角膜内皮细胞Y型连接断裂;而实验B、D组,保存的角膜环轻度水肿、后弹力层皱褶少、角膜透明度高、内皮细胞形态结构完整,超微结构显示保存角膜内皮细胞连接紧密,Y型连接无明显断裂,细胞表面可见较丰富的微绒毛,胞体大,核突起明显。结论bFGF和EGF在角膜中期保存液中均有促进细胞增殖、保持角膜细胞活性的作用,以bFGF作用更明显。Objective To investigate the effect of basic fibroblast growth factor （bFGF） and epidermal growth factor （EGF） on corneal endothelial cell preserved in mid-term preserving medium. Method Corneal rings were preserved in mid-term preserving medium in control group, bFGF（5 ng/ml）, bFGF（20 ng/ml） ,EGF and bFGF ± EGF were added into the mid-term preserving medium in experimental groups A, B, C, D respectively. At the 3rd, 7th and 14th day after preservation, the viability of corneal endothelial cells （CECs） was assessed by trypan blue and alizarin red stain. The microstmcture of corneal endothelial cells was observed by electron microscope （EM）. Results At the 14th days of preservation, viability of CECs was （ 10.35 ± 1.32 ） % （ control group）, （ 62. 18 ± 1.56） % （ group A ）, （ 92.57 ± 0.90）% （group B）, （71.01 ±2.67）% （ group C） and （82.59±1.45）% in group D respectivily. The viability of CECs in experimental groups was much higher than in control group （P 〈 0.01 ）. At the 14th days, corneal rings in control group showed edematous, cloud, defect of integrity of endothelial cells and Y type conjunction broken. But in experimental groups, the corneal edema and folds were slightly, furthermore the cornea ring was transparency and cell shape was intact, especially in group B and group D. Electronic microscope （EM） showed that corneal endothelial cells Y type conjunction in experimental groups was completely integrity, endothelial cells had tight conjunction and rich micro-villa on surface, large cells body and obvious nucleus. Conclusion Addition of bFGF and EGF in corneal storage medium plays a significant role in maintaining CECs structure and viability.

关 键 词：内皮 角膜 内皮细胞 细胞存活 成纤维细胞生长因子2 表皮生长因子 组织保存 

分 类 号：R779.65[医药卫生—眼科]