JAM－1功能缺失对移植角膜透明度的影响

王光洁，蒋　华



JAM－1功能缺失对移植角膜透明度的影响

王光洁，蒋华＊

［摘要］目的观察2种处理方式的供体角膜在移植后的成活情况研究角膜内皮JAM－1的功能。方法首先进行免疫组织化学检测，以确定所用抗体与大鼠角膜内皮细胞的结合。然后处理供体角膜片，24片供体角膜取自12只Wistar大鼠，直径3 mm。其中右眼12片为实验组，左眼12片为对照组。用不含CaCl2和MgCl2的Hank′s液孵育以使细胞连接断开利于抗体结合；之后实验组加入抗JAM－1抗体EP1042Y，对照组加入抗体稀释液0.2 ml，作用15 min。滤纸吸干水分，转入高糖型DMEM，31.7℃培养保存3 d；最后，进行同种异体移植，24只受体大鼠分为两组，分别使用实验组和对照组供体角膜，植床直径3 mm，术后5－0丝线间断缝合上下睑缘2针。术后3、7、10 d进行裂隙灯检查，观察和评估角膜透明度和新生血管情况。结果免疫组织化学结果显示JAM－1分子存在于角膜内皮和上皮细胞，并且能与抗体EP1042Y有效结合。JAM－1功能缺失的供体角膜在各观察时间点透明度均较对照组差并且易于出现新生血管。结论作为细胞紧密连接构成成分的JAM－1，对维持供体角膜的透明和移植的成功必不可少。

［关键词］JAM－1；组织培养保存；角膜移植；紧密连接；角膜内皮细胞

［作者单位］250031山东济南，济南军区总医院眼科（王光洁，蒋华）

Influence of JAM－1 afunction on the transparency of corneal graft

WANG Guang－jie，JIANG Hua.Department of Ophthalmology，the General Hospital of Jinan Military Region，Jinan，Shandong 250031，China

［Abstract］ObjectiveTo approach the function of JAM－1 in corneal endothelium by observing the transparency and the development of neovessels of 2 group corneal grafts processed by different methods before penetrating corneal transplantation.Methods Immunohistochemistry methods were used to detect the presence of JAM－1 in corneal endothelium and the antibody's binding with the cell junctions of corneal endothelium.Twentyfour donor corneas were derived from 12 Wistar rats and the diameter of graft were 3mm.Twelve grafts from the right eye were arranged to experimental group，and rest 12 grafts from the left eye to control group.All corneas were incubated in Hank's solution without CaCl2and MgCl2for 10 min in order to break the cell junctions for pomoting the antidody's binding.Then corneas of the experimental group were incubated with antibody EP1042Y solution and that of control group were incubated with antibody dilution for 15 min.After ward，water content of these corneas were dried with filter paper and them were conserved in DMEM with high glucose for 3 d at 31.7℃；finally allogeneil graft penetrating cornea transplantation was carried out using the 2 group grafts described above.Twenty－four acceptor rats were divided into 2 groups equally for receiving the corneas of experimental group and control group，respectively.The recipient bed was designed to 3mm in diameter.After surgery，the margo palpebraes were sutured with 5－0 suture silk.Slit－lamp examination were processed on the day 3，7 and 10 after surgery，and the transparency and the development of neovessels were observed and evaluated.Results Immunohistochemistry showed that JAM－1 resided in the cell junction of cornea endothelium and epithelium and that the used antibody EP1042Y was effective.In cornea transplantation，the JAM－1 afunction grafts were less transparent than the control group and prone to develop neovessels.Conclusion JAM－1，as a component of cell junctions，it's function is essential for corneal grafts' transparency and even for the success of penetrating cornea transplantation surgery.

［Key words］JAM－1；Tissue culture preservation；Corneal transplantation；Corneal endothelium cell

上皮和内皮组织中，接合黏附分子－1（junctional adhesion molecule－1，JAM－1）是紧密连接有调节作用的跨膜分子。其通过N末端环的同嗜性黏附调节紧密连接的渗透性起作用。有资料显示角膜内皮细胞的紧密连接是渗透性屏障主要功能区，对维持角膜的透明有重要作用，而JAM－1分子是这一屏障的重要构成组分。……