Edelhauser H F, Van Horn D L, Miller P, Pederson H J
J Cell Biol. 1976 Mar;68(3):567-78. doi: 10.1083/jcb.68.3.567.
Intracellular-reduced glutathione (GSH) was removed by thiol-oxidation with diamide during in vitro perfusion of the corneal endothelium. By 15 min the normal mosaic-like pattern of the endothelial cells was disrupted by serpentine-like lines of cell separation at the cell juntions. After 45 min of perfusion, infividual clusters of cells formed cup-shaped islands. The resultant exposure of Descemet's membrane to the perfusion solution resulted in corneal swelling. Transmission electron microscopy revealed that the endothelial cells separated at the apical junctions and that the microfilaments in the apical cytoplasm of cells formed dense bands, whereas the other subcellular organelles were normal in appearance. The change in cellular shape may be due to loss of cellular adhesion which results in the condensation of the microfilaments or contraction of the microfilaments. The addition of glucose to the perfusate prevented the diamide effect, and the diamide effect could be reversed upon removal and perfusion of a glutathione bicarbonate Ringer's solution. These results suggest that the ratio of reduced to oxidized glutathione in the endothelial cells plays a role in the maintenance of the endothelial cell barrier function.
在角膜内皮细胞的体外灌注过程中，通过用二酰胺进行硫醇氧化去除细胞内还原型谷胱甘肽（GSH）。到15分钟时，内皮细胞正常的镶嵌样模式在细胞连接处被蛇形的细胞分离线破坏。灌注45分钟后，单个细胞簇形成杯状岛。Descemet膜暴露于灌注溶液导致角膜肿胀。透射电子显微镜显示，内皮细胞在顶端连接处分离，细胞顶端细胞质中的微丝形成致密带，而其他亚细胞细胞器外观正常。细胞形状的变化可能是由于细胞黏附丧失，导致微丝凝聚或微丝收缩。向灌注液中添加葡萄糖可防止二酰胺效应，并且在去除并灌注谷胱甘肽碳酸氢盐林格氏液后，二酰胺效应可逆转。这些结果表明，内皮细胞中还原型谷胱甘肽与氧化型谷胱甘肽的比例在维持内皮细胞屏障功能中起作用。
