接合粘附分子一1功能缺失对大鼠角膜内皮的影响

目的 通过阻断接合粘附分子-l引起角膜内皮细胞结构和活性的改变来研究JAM-1的作用.方法 由14只大鼠取下28片3.5mm直径角膜片,其中右眼角膜为实验组,用抗JAM-1单克隆抗体处理,左眼角膜为对照组,用磷酸缓冲液处理.处理后在高糖DMEM液中进行培养保存5d,之后用含5％葡聚糖T500的DMEM脱水24 h.脱水后每组有12片角膜做锥蓝-茜素红染色内皮细胞活性计数及厚度测量,2片角膜与另外2片新鲜角膜一同进行戊二醛固定,用于透射电子显微镜观察.结果 活性细胞计数对照组为(2008±505 )/mm2,实验组为(934±521 )/mm2,P＜0.0l.角膜厚度对照组为(375.02±83.33)μm,实验组为(461.81±39.14)μm,P＜0.01.超微结构显示,实验组内皮细胞损坏严重,有较多的自噬体形成.结论 作为细胞紧密连接构成成份的JAM-1,对维持角膜的内皮细胞结构和活性必不可少.

Effect of JAM-1 blockage on culture storage rat cornea endothelium

Objective To approach the function of JAM-1 in corneal endothelium by observing cornea endothelium structure and activity change induced by antibody blockage.Methods Twenty-eight corneas were obtained from 14 rats and were divided into two groups.Corneas of the right eye were in experiment group and processed with monoclonal antibody of JAM-1 to block the function of JAM-1; corneas of the left eye were in control group and processed with PBS buffer.All corneas were cultured in DMEM with high glucose for 5 days and then dehydrated in DMEM containing 5％dextran-T500 for 24 hours.Twelve corneas of each group were underwent Trypan blue - alizarin vital staining and cornea thickness were measured.Two corneas of each group and other 2 fresh normal corneas were fixed with 4％ glutaraldehyde for transmission electron microscopy.Results The active cell counting of experiment group and control group were (2008± 505)/mm2 and (934± 521 )/mm2 respectively,P ＜0.01.The cornea thickness of the two groups were (375.02± 83.33)μ m and (461.81±39.14)μ m respectively,P ＜0.01.Transmission electron microscopy showed that in experiment group the endothelium cell appeared more ultrastructural damage and autophagosome formation.Conclusions JAM-l,as a component of cell junctions,its function is essential for corneal endothelium structure and activity in tissue culture preservation.