去瓣型Epi-LASIK与LASEK术后角膜上皮内紧密连接蛋白occludin表达的实验研究

发布时间：2018-04-11 04:34

本文选题：角膜磨镶术 + 上皮下　；参考：《安徽医科大学》2012年硕士论文

【摘要】：目的:通过对兔去瓣型Epi-LASIK与LASEK术后早期角膜组织的研究，探讨两种手术方式各自对于角膜上皮内紧密连接蛋白occludin的影响。方法：48只新西兰兔按不同取材时间24,48,72,120小时随机分为4组，每组12只实验兔，其中6只实验兔右眼行去瓣型Epi-LASIK，另外6只右眼20%酒精浸润角膜后行LASEK术，，亦不保留上皮瓣，另有2只兔为空白对照组。术后24,48.72.120小时空气栓塞法处死动物分别取材， OCT冰冻包埋制作冰冻切片，行HE染色明确角膜上皮修复情况，免疫荧光染色检测观察角膜上皮内紧密连接蛋白occludin的变化，RT-PCR半定量检测中央区角膜内occludin蛋白mRNA表达的变化，对数据进行统计学分析。结果 1.组织形态学改变：HE染色示两组兔眼角膜上皮缺损在术后2天均已被新生角膜上皮覆盖，但与正常上皮相比较薄，细胞数较少，术后3天角膜上皮基本恢复正常形态。 2.免疫荧光染色检测：正常角膜中，occludin蛋白主要表达于角膜横侧面的表浅细胞层中，组成排列致密连续的紧密连接结构。术后24h，48h，72h去瓣型Epi-LASIK组角膜上皮occludin蛋白的修复速度及表达水平均高于去瓣型LASEK组，术后120h两实验组角膜上皮内occludin蛋白结构均基本修复完毕，与正常对照组之间无明显差异。3.RT-PCR检测：术后24h，48h，72h去瓣型Epi-LASIK组与LASEK组中央角膜内occludin蛋白mRNA的表达差异具有统计学意义（P0.05），术后120h两组之间的表达差异无统计学意义（P0.05）。结论：去瓣型Epi-LASIK相对于LASEK手术对于角膜上皮紧密连接的破坏更轻，术后角膜上皮紧密连接的修复也更加迅速，提示其有利于减少术后角膜刺激症状和并发症。
[Abstract]:Aim: to investigate the effects of the two methods on the intraepithelial tight junction protein (occludin) in rabbit cornea after Epi-LASIK and LASEK.Methods Twenty eight New Zealand rabbits were randomly divided into 4 groups according to the time of different sampling time: 12 rabbits in each group. Six of them were treated with Epi-LASIKK in the right eye, and the other 6 eyes were treated with LASEK after 20% alcohol infiltration of cornea, and the flap was not preserved.Another 2 rabbits were used as blank control group.The animals were killed by air embolization for 24 hours 48.72.120 hours after operation. The frozen sections were made by OCT frozen embedding, and the corneal epithelium repair was determined by HE staining.The changes of occludin in corneal epithelium were detected by immunofluorescence staining. The expression of occludin protein mRNA in central cornea was detected by semi-quantitative RT-PCR, and the data were analyzed statistically.Result1.Histological changes: he staining showed that the corneal epithelial defects of the two groups had been covered by neonate corneal epithelium 2 days after operation, but the corneal epithelium was thinner than the normal epithelium, and the number of cells was less, and the corneal epithelium returned to normal shape 3 days after operation.2.Immunofluorescence staining showed that the protein of cludin was mainly expressed in the superficial cell layer on the lateral side of the cornea and formed a compact and continuous tight junction structure.The repair speed and expression level of occludin protein in corneal epithelium in Epi-LASIK group were higher than those in LASEK group at 24 h, 48 h and 72 h after operation, and the occludin protein structure in corneal epithelium of two experimental groups was basically repaired at 120 h after operation.There was no significant difference between the control group and the control group. 3. RT-PCR: the expression of occludin protein mRNA in the central cornea of the Epi-LASIK group and the LASEK group was significantly different from that of the control group at 24 h, 48 h and 72 h after operation, but there was no significant difference in the expression of occludin protein between the two groups at 120 h after operation (P 0.05).Conclusion: compared with LASEK, the devalved Epi-LASIK has less damage to the tight junction of corneal epithelium, and the repair of the tight junction of corneal epithelium is more rapid after operation, which suggests that it is helpful to reduce the symptoms and complications of corneal irritation after operation.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R779.63

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