人羊膜匀浆上清液治疗兔角膜碱烧伤病理研究.docVIP

人羊膜匀浆上清液治疗兔角膜碱烧伤病理研究 　　【摘要】目的:观察人羊膜匀浆上清液促进兔角膜碱烧伤后角膜上皮和基质修复的作用,为临床上应用人羊膜匀浆上清液点眼治疗眼碱烧伤提供理论依据#65377;方法:将制备的羊膜匀浆上清液点眼治疗兔角膜碱烧伤,并对治疗后的角膜照相观察角膜上皮愈合和基质修复情况#65377;结果:羊膜匀浆上清液治疗角膜碱烧伤后眼充血明显减轻,角膜上皮愈合较快,角膜上皮细胞排列整齐,基质中炎症细胞浸润减轻且胶原纤维排列整齐#65377;结论:早期应用羊膜匀浆上清液能促进兔角膜碱烧伤后角膜的愈合及膜基质有序修复#65377; 　　【关键词】羊膜匀浆上清液;角膜;碱烧伤 　　 文章编号:1009-5519(2007)11-1582-02中图分类号:R77文献标识码:A 　　Patho-study on human amniotic homogenate to corneal healing after alkali burn of cornea in rabbits 　　LIAO Qiong,DAI Chao,LIU Xiang 　　(Department of Ophthalmology,Xinqiao Hospital,The Third Military University of Medical Sciences,Chongqing,400037,China) 　　【Abstract】Objective:To select the effective methods for eye alkaline fresh amnion to rabbit corneal epithelial cell and stroma after alkali burn of rabbit cornea.Methods:The model of alkali burn of rabbit cornea was set up and treated by supernatant of homogenized fresh amnion.Corneal epithelium and stroma were observed with a slit lamp microscope.Results:After treatment with supernatant of homogenized fresh amnion,the rabbit eyes showed progressive decrease of vascularity and stromal infiltration and orderly arrangement of corneal epithelial cell and collagen fibers.Conclusion:Supernatant of homogenized fresh amnion can accelerate the healing of rabbit corneal epithelium and restore the corneal stroma in order. 　　【Key words】Supernatant of homogenized amnion;Cornea;Alkali burn 　　 　　文献报道羊膜上皮细胞分泌EGF#65380;TGF-a#65380;KGF#65380;HGF#65380;bFGF等生长因子[1,2],我们直接将羊膜制成匀浆,观察其存在的多种细胞因子是否对角膜上皮细胞的增殖有促进作用,并将羊膜匀浆上清液制成眼液点眼治疗兔角膜碱烧伤,然后行角膜病理研究,观察其对角膜上皮愈合#65380;炎症反应#65380;基质修复的影响,以期寻找一种比羊膜移植经济#65380;方便#65380;效果佳的治疗角膜碱烧伤的??方法#65377; 　　 　　1 资料和方法 　　 　　1.1 动物及主要试剂:(1)家兔:体重1.5~2.0 kg#65377;(2)羊膜匀浆上清液:自制,方法如下:在超净台上将羊膜在PBS液中清洗3次,约10~15分钟,取适当大小称湿重,按1∶1加PBS液放入匀浆机中,每分钟5 000转,10冲程磨制匀浆,每冲程30秒#65377;取出匀浆按1∶5加PBS液于离心管中离心2 000 r/min,10分钟,取上清液备用(1小时内用)#65377;后加入细胞冻存液放入-80℃冰箱过夜,再放入液氮中冻存#65377;(3)bFGF(贝复舒眼液);(4)1 mol/L NaOH:自制#65377; 　　1.2 碱烧伤动物模型的制备:(1)1%丁卡因眼液点眼3次#65377;(2)乙醚吸入麻醉#65377;(3)开睑器开睑,将浸