穿透角膜移植术后虹膜睫状体血管通透性变化  

Diversity of vascular permeability in iris and ciliary body after penetrating keratoplasty

作　　者：刘明娜[1] 庄宪丽[1] 高华[1] 李素霞[1] 史伟云[1] 

机构地区：[1]山东省眼科研究所山东省眼科医院,济南250021

出　　处：《中华实验眼科杂志》2013年第7期636-641,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目（81170815）;山东省自然科学基金项目（ZR2012HQ041）

摘　　要：背景穿透角膜移植术后发生排斥反应是手术失败的主要原因，尤其是存在高危排斥反应条件的患者。以往对角膜移植排斥反应发生机制的研究主要集中于角膜缘和角膜新生血管（CNV）方面，但目前发现，这些因素并不能解释所有角膜植片排斥现象。研究发现，虹膜睫状体组织内存在免疫细胞，但其与角膜植片排斥反应的关系尚不清楚。目的研究穿透角膜移植术后虹膜睫状体的血管变化情况。方法将8周龄清洁级BALB／c小鼠70只分为同种异体角膜移植组60只和空白对照组10只。角膜移植组以C57BL／6小鼠角膜作为供体，BALB／c小鼠作为受体，行穿透角膜移植术。术后每天裂隙灯显微镜下观察并记录角膜植片混浊情况，并按Hegde的标准进行评分。分别于术后5d、10d和发生排斥时选取各组小鼠，处死后收集虹膜睫状体标本，分别应用免疫组织化学法和逆转录PCR（RT-PCR）法检测虹膜睫状体中跨膜蛋白occludin、胞质附着蛋白-1（ZO-1）、基质金属蛋白酶-9（MMP-9）、主要组织相容性复合体Ⅱ（MHC1I）、趋化因子受体（CCR5、CCR7）及其mRNA在各组小鼠虹膜睫状体组织中的表达，用ImageJ图像分析软件计算虹膜睫状体组织铺片中上述各因子阳性细胞的数量（个／视野）以及RT—PCR中目的基因的吸光度（A）值。结果角膜植片平均存活时间为（17±3）d，术后5d角膜植片评分平均为0．6，术后10d平均评分为0．5，术后18d平均评分为3．3。正常小鼠虹膜睫状体组织中可见大量ZO-1蛋白表达，排斥反应时可见ZO-1蛋白表达明显减少。正常小鼠虹膜睫状体组织中未见MMP-9的表达，角膜移植术后免疫排斥反应时可见大量MMP-9的表达，呈红色荧光。虹膜睫状体组织铺片免疫荧光染色显示，角膜移植术后免疫排斥反应时MHCII＋细胞数量较正常小鼠明显增加，分别为（4021．83±495．18）和�Background Allograft rejection is a main cause of failure of penetrating keratoplasty, especially in the patient with high risk of rejection condition. Previous study on allograft rejection mechanism focused on limbal and corneal neovascularization, but these factors did not explain all the phenomena of allograft rejection. Research found that immune cells appeared in iris and ciliary body when rejection occurred, but the relationship between these immune cells and allografi rejection is unclear Objective This study was to evaluate the relationship between diversity of vascular permeability in the iris and ciliary body and allograft rejection after penetrating keratoplasty. Methods Seventy clean eight-week-old BALB/c mice were divided into allogeneic corneal transplantation group （60 mice） and blank control group （10 mice）. Allogeneic corneal transplantation was performed with the same age of C57BL/6 mice as donor and BALB/e mice as the recipients. The grafts were examined under the slit lamp microscope and scored based on the criteria of Hegde. The mice were sacrificed and iris and ciliary tissue were obtained 5, 10 days and rejection after surgery. Immunohistochemistry and reverse transcription PCR （RT-PCR） was used respectively to detect the expression diversities of occludin, zonula occludens protein-1 （ZO-1）, matrix metalloproteinase-9 （MMP-9） , major histocompatibility complex- H （ MHC- H ） , and CCRS, CCR7 and their mRNA in iris and ciliary body. Image-J image analysis software was used to calculate the quantity of positive cells on iris wholemount, and absorbance of target genes （A values）. The use and care of the experimental animals complied the ARVO Resolution on the Use of Animals in Research. Results The mean survival time of corneal grafts was （ 17±3） days after operation. The mean score was 0.6 in 5 days and 0.5 in 10 days, and 3.3 in 18 days after operation. Expression of ZO-1 reduced significantly, and that of MMP-9 increased obviously at the time of rejection. MH

关 键 词：穿透角膜移植 免疫排斥 虹膜睫状体 血管通透性 

分 类 号：R779.65[医药卫生—眼科]