羊膜提取液抑制兔Epi-LASIK术后haze形成的作用和机制  

Study on the mechanism of amniotic extraction inhibiting haze formation after Epi-LASIK in rabbit cornea

作　　者：夏世刚[1] 王俞方[1] 肖启国[1] 彭华[2] Shi-Gang Xia Yu-Fang Wang Qi-Guo Xiao Hua Peng(Department of Ophthalmology Department of Otolaryngology, the Second Hospital, University of South China, Hengyang 421001, Hunan Province, China)

机构地区：[1]南华大学附属第二医院眼科,中国湖南省衡阳市421001 [2]南华大学附属第二医院耳鼻喉科,中国湖南省衡阳市421001

出　　处：《国际眼科杂志》2017年第8期1427-1432,共6页International Eye Science

基　　金：湖南省卫生厅科研计划项目(No.B2010-052)~~

摘　　要：目的:探讨羊膜提取液(AE)抑制兔微型角膜刀法准分子激光角膜上皮瓣下磨镶术(Epi-LASIK)后角膜上皮下雾状混浊(haze)形成的作用和机制。方法:新西兰白兔36只72眼按照随机数字表法分为三组:AE组、激素组和溶媒对照组。构建Epi-LASIK动物模型,术后AE组滴用AE眼液,激素组滴用1g/L磷酸地塞米松眼液,溶媒对照组滴用不含AE的溶媒眼液。术后1、4、8wk在裂隙灯下进行haze分级评估。HE染色观察角膜上皮的修复,免疫组织化学法观察角膜上皮细胞NF-kB P65蛋白的表达。用ELISA法检测炎症细胞因子(TNF-α、TGF-β1和IL-1β)和抗炎症细胞因子(IL-4、IL-10和IL-13)的表达水平。结果:HE染色结果显示,与激素组和溶媒对照组比较,Epi-LASIK术后1wk,AE组兔角膜上皮基底细胞形态较均匀,排列较规则;术后4wk时,AE组浅基质层内出现少量新生胶原纤维,排列较规则,形成少量瘢痕;术后8wk,AE组角膜浅基质层见少量新生胶原纤维,排列规则,极少形成瘢痕。在术后早期(1wk和4wk),AE组与激素组、溶媒对照组相比,具有明显抑制炎症因子(TNF-α、TGF-β1和IL-1β)和促进抗炎症因子(IL-4、IL-10和IL-13)分泌的作用,组间比较差异有显著统计学意义(P<0.01)。而且,在术后早期(1wk和4wk),AE组与激素组、溶媒对照组相比,角膜上皮细胞NF-kB通路关键组份P65蛋白浅染,细胞核淡染,明显抑制NF-kB信号通路的激活。结论:羊膜提取液可能通过抑制角膜上皮细胞的NF-kB信号通路,减轻炎症反应,从而减少胶原与瘢痕形成和haze的发生。AIM： To investigate the probably mechanism of amniotic extraction inhibiting haze formation after epipolis laser in situ keratomileusis （Epi-LASIK） in rabbit cornea.METHODS： Thirty rabbit corneas were performed with Epi-LASIK.All eyes were randomly divided into three groups： eyes treated with amniotic extraction （AE group）,eyes treated with 1g/L dexamethasone （hormone group） and eyes treated with solvent （solvent control group）.Haze grade evaluation was performed under the slit lamp after Epi-LASIK for 1,4 and 8wk.The repair of corneal epithelium was observed by using HE staining,and the expression of NF-kB protein P65 was detected by immunohistochemistry.The expression levels of inflammatory cytokines （TNF-α,TGF-β1 and IL-1） and anti-inflammatory cytokines （IL-4,IL-10 and IL-13） were determined by ELISA.RESULTS： HE staining showed that the basal cells of corneal epithelium were more uniform and arranged regularly in AE groups after Epi-LASIK for 1wk as compared with the hormone group and the solvent control group.After 4wk,there were a few of new collagen fibers in the superficial stroma of AE group,forming a small amount of scar.After 8wk,the corneal stroma of AE group showed a small amount of new collagen fibers,arranged regularly,and rarely formed scar.At the early stage （1 and 4wk）,AE treatment has an obviously effect on inhibiting the secretion of inflammatory factors （TNF-α,TGF-β1 and IL-1） and anti-inflammatory factors （IL-4,IL-10 and IL-13）,and the difference was statistically significant （P〈0.01）.Moreover,the activation of the NF-kB signaling pathway was significantly inhibited by treatment with AE in the early postoperative period （1 and 4wk）.CONCLUSION： Amniotic extraction may reduce the inflammatory response in corneal epithelial cells by inhibiting the NF-kB signaling pathway,thereby inhibiting the formation of collagen and scar and the occurrence of haze.

关 键 词：羊膜提取液 微型角膜刀法准分子激光角膜上皮瓣下磨镶术 角膜混浊 NF-kB通路 炎症细胞因子 

分 类 号：R778[医药卫生—眼科]