硫酸软骨素在甘油冷冻液中对角膜内皮保护作用的实验研究  

Experimental study of the chondroitin sulfate in glycerol freezing solution on the protection of corneal endothelium

作　　者：王宏伟[1] 邱姣[1] 赵青[1] 

机构地区：[1]辽宁医学院附属第一医院眼科,中国辽宁省锦州市121001

出　　处：《国际眼科杂志》2011年第9期1525-1527,共3页International Eye Science

基　　金：中国辽宁省教育厅高等学校科学研究资助项目(No.2009A445)~~

摘　　要：目的:探讨硫酸软骨素(chondroitin sulfate,CS)在甘油冷冻保存角膜中对角膜内皮细胞(corneal epithelial cells,CEC)的保护作用。方法:Wistar大鼠角膜片40只随机分为实验组和对照组,对照组角膜片直接放入纯甘油中,实验组则在内皮面均匀黏附10g/L CS后再放入甘油中。-20℃保存2mo后行CEC台盼蓝-茜素红活性染色及电镜检查。取30只SD大鼠与保存后角膜片建立Wistar-SD同种异体穿透性角膜移植模型,测定术后不同时间点植片内TNF-α和IFN-γ的表达水平,检验保存效果。结果:台盼蓝-茜素红活性染色结果及术后植片内TNF-α和IFN-γ的表达结果显示,两组差异有显著性(P<0.05),实验组保存效果和角膜移植术后效果均优于对照组。电镜检查示,实验组CEC胞质内可见个别空泡,线粒体及内质网轻度肿胀;对照组CEC线粒体及内质网明显肿胀。结论:甘油冷冻能够保持CEC的活性,且CS能进一步增进保存效果。AIM：To evaluate chondroitin sulfate on the glycerol cryopreservation of corneal endothelial cells（CEC）.METHODS：Forty Wistar rats corneal pieces were divided into the experimental group：glycerol cryopreservation medium containing chondroitin sulfate（CS）;and the control group：mere glycerol cryopre-servation medium.The two groups were saved at-20℃ for 2 months.All pieces were examined with trypan blue combined with alizarin red staining（viability staining） and electron-microscope examination for CEC.30 SD rats were taken to establish Wistar-SD rat model of allogenic penetrating keratoplasty.Slit lamp microscope detected the occurrence of corneal rejection,TNF-α and IFN-γ expression was examined by immunohistochemistry to test the effect of preservation.RESULTS：The staining results,electron-microscope examination and TNF-α,IFN-γ expression showed,the difference was significant in these two groups（P0.05）,the results of the experimental group was better than the control group after the saving and the corneal transplantation（P0.05）.The electron-microscope examination showed indivdual vacuoles in CEC cytoplasm,slight swelling of mitochondrial and endoplasmic reticulum in experimental group,obvious swelling of CEC mitochondrial and endoplasmic reticulum in control group.CONCLUSION：Glycerol cryopreservation can preserve the viability of CEC.Chondroitin sulfate can obviously improve the effect of preservation.

关 键 词：硫酸软骨素 甘油冷冻 角膜 角膜内皮细胞 

分 类 号：R779.65[医药卫生—眼科]