Ⅱ型胶原酶构建兔角膜离体扩张模型  

Creating a rabbit model for corneal ectasia in vitro with collagenase type Ⅱ

作　　者：乔静[1] 李海丽[1] 宋文静[1] 汤韵[1] 荣蓓[1] 杨松霖[1] 吴元[1] 晏晓明[1] 

机构地区：[1]北京大学第一医院眼科视觉损伤与修复教育部重点实验室,100034

出　　处：《中华眼视光学与视觉科学杂志》2016年第5期275-279,共5页Chinese Journal Of Optometry Ophthalmology And Visual Science

基　　金：基金项目：国家自然科学基金（11372011）;北京市自然科学基金（7142159）;非线性力学国家重点实验室开放基金

摘　　要：目的 探索使用Ⅱ型胶原酶建立兔角膜离体扩张模型的方法及其可行性.方法 实验研究.20只离体兔角膜采用随机数字表法分为Ⅰ组（阴性对照组）、Ⅱ组（5 mg/mlⅡ型胶原酶组）、Ⅲ组（10 mg/mlⅡ型胶原酶组）及Ⅳ组（15 mg/mlⅡ型胶原酶组）,每组5只角膜.刮除上皮后角膜置于人工前房上,各组采用不同浓度Ⅱ型胶原酶溶液浸泡60 min.分别记录处理前后各组角膜在不同人工前房内压力[模拟眼压（I0P）]下（15、30、45 mmHg）角膜平均曲率及角膜中央厚度,并对处理后各组角膜进行组织学检查.各组处理前后角膜平均曲率差值△Km及角膜中央厚度差值△CCT的变化采用单因素方差分析.结果 不同IOP梯度下,4组角膜处理前后角膜平均曲率差值△Km差异有统计学意义（F=8.46、9.24、8.58,P＜0.01）;在各个IOP梯度下各组比较发现Ⅳ组与Ⅰ组的△Km差异均有统计学意义（P＜0.01）,Ⅱ组、Ⅲ组与Ⅰ组的△Km差异均无统计学意义（P＞0.05）.不同IOP梯度下,4组角膜处理前后角膜中央厚度差值△CCT差异均无统计学意义（F=0.22、0.66、1.60,P＞0.05）.与阴性对照组相比,胶原酶处理过的角膜基质胶原纤维排列疏松,基质水肿.结论 15 mg/mlⅡ型胶原酶溶液浸泡角膜60min能使角膜平均曲率增加,可用于建立兔角膜离体扩张模型.Objective To investigate how to develop a rabbit model for corneal ectasia in vitro with collagenase type Ⅱ.Methods In this experimental study,20 postmortem New Zealand white rabbit corneas were randomly divided into 4 groups：group Ⅰ （the control group with 15% dextran）,group Ⅱ （5 mg/ml collagenase type Ⅱ with 15% dextran）,group Ⅲ （10 mg/ml collagenase type Ⅱ with 15% dextran） and group Ⅳ （15 mg/ml collagenase type Ⅱ with 15% dextran）.After epithelial debridement,corneas were mounted and pressed onto artificial anterior chambers and different kinds of solutions were applied to the corneas for 60 minutes.The average corneal curvature and central comeal thickness were measured before and after collagenase exposure at different intraocular pressure （IOP） levels （15 mmHg,30 mmHg,45 mmHg）.Corneal specimens were processed for light microscopy with HE staining.Changes in average corneal curvature and central corneal thickness were analyzed statistically by one-way ANOVA in SPSS 17.0.Results After collagenase exposure,significant changes in △Km （F=8.46,9.24,8.58,P〈0.01） were seen among groups at 15 mmHg,30 mmHg and 45 mmHg.A significant increase in △Km was seen in group Ⅳ across all IOP levels （P〈0.01） compared to group Ⅰ,while no significant changes were seen in groups Ⅱ and Ⅲ （P〉0.05）.No significant changes in △CCT were seen among groups across all IOP levels （F=0.22,0.66,1.60,P〉0.05）.Light microscopy showed a loss of the normal lamellar structure and changes in collagen organization after collagenase exposure.Conclusion 15 mg/ml collagenase type Ⅱ can induce a significant increase in average corneal curvature,which might be considered for a method of building the rabbit model for corneal ectasia in vitro.

关 键 词：基质金属蛋白酶8 圆锥角膜 兔 角膜扩张 角膜曲率 角膜厚度 Matrix metallo PROTEINASE 8 

分 类 号：R-332[医药卫生] R772.2