胶原交联对兔角膜碱烧伤新生血管影响的实验研究

发布时间：2018-04-14 22:32

本文选题：角膜胶原交联术 + 角膜新生血管　；参考：《遵义医学院》2017年硕士论文

【摘要】：目的:本研究拟通过观察兔眼碱烧伤诱导角膜新生血管模型经核黄素联合紫外光A行角膜胶原交联(corneal collagen cross linking,CXL)后新生血管的生长,探索CXL对角膜新生血管的影响及机制。方法:健康家兔60只,随机分为6组,每组10只,右眼为实验眼;A组为单纯碱烧伤对照组,B、C、D、E、F组为实验组,其中B组为碱烧伤后即刻标准CXL(标准CXL方法为紫外光光照30分钟)、C组为碱烧伤后即刻CXL紫外光光照时间15分钟、D组为碱烧伤后即刻CXL紫外光光照时间45分钟、E组为碱烧伤后1天标准CXL、F组为碱烧伤后3天标准CXL,表麻下予1 mol/L的氢氧化钠建立碱烧伤模型及胶原交联干预各组模型。碱烧伤模型建成后分别于第4、7、10、14天观察并记录角膜新生血管面积、角膜水肿、感染、溃疡,于第7、14天分别各取5只家兔角膜组织,采用Western-Blotting分析VEGF、MMP-2、MMP-9、TIMP-1的表达量。结果:1.术后裂隙灯观察:第14天处死各组家兔,单纯碱烧伤对照组10眼中有5只眼(50%)发生角膜中央基质溶解、后弹力层膨出;实验组共50只眼,其中有7只眼(14%)出现角膜中央变薄,3只眼(6%)出现角膜溶解,各组均未发生角膜穿孔。2.角膜新生血管(CRNV)面积比较:第4天:各组间比较无显著差异(P0.05);第7天:各组间比较有显著差异(P0.05),其中以A组CRNV面积最大、D组CRNV面积最小;第10天:A与D组、A与E组间相比有显著差异(P0.05),D、E组CRNV面积较A组显著减小。第14天:A与D组、B与D组间及F与D组间相比较有显著差异(P0.05),D组较A、B、F组的CRNV面积显著减小。3.Werstern-Blotting检测VEGF、MMP-2、MMP-9和TIMP-1表达量的比较:在兔眼角膜碱烧伤后第7天,对照组与各实验组之间VEGF、MMP-2在角膜组织中的表达量无显著差异(P0.05);MMP-9的表达量在A组显著低于各个实验组(P0.05);TIMP-1的表达量比较中A组显著低于D、F组(P0.05);在角膜碱烧伤后第14天,对照组A组角膜组织中VEGF、MMP-2、MMP-9和TIMP-1的表达量均较前上升,其中对照组A组VEGF表达量显著高于各个实验组(P0.05),A组角膜中MMP-2表达量显著高于B、D、E组(P0.05);A组角膜组织中MMP-9、TIMP-1表达量均高于各个实验组,但未达到显著差异水平(P0.05)。结论:1.本研究进一步证实了紫外光A联合核黄素胶原交联具有抗角膜基质溶解的作用;2.紫外光A联合核黄素胶原交联45分钟可以减轻角膜新生血管的形成,其可能通过下调VEGF、MMP-2、MMP-9的表达而抑制角膜新生血管生长。
[Abstract]:Objective: to investigate the effect and mechanism of CXL on corneal neovascularization by observing the growth of corneal neovascularization induced by alkali burn in rabbits by riboflavin combined with ultraviolet light A (UV).Methods: sixty healthy rabbits were randomly divided into 6 groups (n = 10 in each group).Group B was the standard CXLimmediately after alkali burn (the standard CXL method was UV illumination for 30 minutes, group C was UV light time immediately after alkali burn for 15 minutes, group D was for 45 minutes CXL ultraviolet illumination time immediately after alkali burn, group E was treated with ultraviolet light for 30 minutes after alkali burn).The model of alkali burn was established 1 day after alkali burn with sodium hydroxide of 1 mol/L in standard CXLF group 3 days after alkali burn and collagen crosslinking was used to interfere each group model.The corneal neovascularization area, corneal edema, infection and ulcer were observed and recorded on the 4th day, 7th day, 10th day after the establishment of alkali burn model. The corneal tissues of 5 rabbits were taken respectively on day 714, and the expression of TIMP-1 was analyzed by Western-Blotting.The result is 1: 1.Postoperative slit lamp observation: the rabbits of each group were killed on the 14th day, 5 eyes of 10 eyes of the control group were treated with simple soda burn, and 50 eyes of the control group were treated with corneal central stroma dissolution and posterior elastic layer swelling, 50 eyes of the experimental group were observed.There were 7 eyes with central corneal thinning, 3 eyes with central corneal thinning and 6 eyes with corneal dissolution, and no corneal perforation occurred in all groups.Comparison of corneal neovascularization area: on the 4th day, there was no significant difference among the three groups (P 0.05), and on the 7th day, there was significant difference between the groups (P 0.05). The CRNV area of group A was the largest and that of group D was the smallest.On the 10th day, there was a significant difference between group A and group D in the area of CRNV between group A and group E. the area of CRNV in group E was significantly lower than that in group A.On the 14th day, there were significant differences between group B and D and between group F and group D. The area of CRNV in group D was significantly reduced than that in group A and group D. Werstern-blotting was used to detect the expression of VEGF, MMP-2, MMP-9 and TIMP-1: on the 7th day after corneal alkali burn, the expression of VEGF MMP-2, MMP-9 and TIMP-1 in rabbit cornea were measured by Werstern-blotting.There was no significant difference in the expression of VEGF MMP-2 in corneal tissue between the control group and the experimental groups. The expression of VEGF MMP-2 in group A was significantly lower than that in group A (P 0.05) and in group A was significantly lower than that in group D (P 0.05), and on the 14th day after corneal alkali burn, the expression of VEGF MMP-2 in group A was significantly lower than that in group D (P 0.05).In group A, the expression of VEGF, MMP-2, MMP-9 and TIMP-1 in corneal tissue were increased.The expression of VEGF in group A was significantly higher than that in group A (P 0.05). The expression of MMP-9 and TIMP-1 in cornea of group A was significantly higher than that of group A (P 0.05), but the expression of TIMP-1 in cornea of group A was significantly higher than that of group A (P 0.05), but the expression of TIMP-1 in cornea of group A was significantly higher than that of group A (P 0.05).Conclusion 1.This study further confirmed that UV A combined with riboflavin collagen crosslinking can inhibit corneal stroma dissolution.Ultraviolet A combined with riboflavin collagen crosslinked for 45 minutes can reduce corneal neovascularization, which may inhibit corneal neovascularization by down-regulating the expression of VEGF MMP-2 and MMP-9.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R779.1

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