CHANGES IN NUCLEAR-DNA CONTENT AND CELL-SIZE OF INJURED HUMAN CORNEAL ENDOTHELIUM
CHANGES IN NUCLEAR-DNA CONTENT AND CELL-SIZE OF INJURED HUMAN CORNEAL ENDOTHELIUM
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DOI：
10.1016/0014-4835(88)90004-8
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发表时间：
1988-08-01
影响因子：
3.4
通讯作者：
FUJITA, S
中科院分区：
文献类型：
--
作者：
IKEBE, H;TAKAMATSU, T;FUJITA, S
为了了解人角膜内皮如何补偿细胞损失，对受损的角膜内皮进行了核 DNA 细胞荧光测定和细胞形态测定。检查角膜：圆锥角膜并发急性积水2例，无急性积水1例；疱疹性角膜炎2例；白内障囊内摘除术后（ICCE后）1例；黄斑角膜炎1例。将内皮细胞层与后弹力层分离并用罗丹明标记的麦芽凝集素-凝集素(WGA)和4'',6-二脒基-2-苯基吲哚二盐酸盐(DAPI)进行双重染色。用彩色图像分析仪测量每个细胞的面积，并与其细胞荧光核DNA含量进行比较。患病角膜表面完好区域的内皮显示出与生理角膜相同的 DNA 倍体模式和细胞面积。然而，即使是二倍体，受损区域的内皮细胞也比正常的内皮细胞面积更大，并且显示出更多数量的从 4C 到 36C 的超倍体细胞。超倍体细胞由许多多核和少量多倍体组成，并且具有极大且奇异的细胞质。所有受伤的角膜都伴有带有大量微核的细胞。 ICCE 后的情况中出现了一些不对称的 4C 双核（DNA 值例如 1.3 加 2.6C）。结论是，体内人角膜内皮细胞的损伤会导致细胞增大，无论是否有 DNA 合成。这些变化在患病的角膜中比在我们之前报道的生理老化的情况下显得更严重。在严重的病例中，微核、多倍体细胞和多核巨细胞很常见，从而表明在角膜严重损伤后，内皮可能存在长期持续的代谢损伤。
To understand how human corneal endothelium compensates for cell loss, nuclear DNA-cytofluorometry and cell morphometry were carried out on injured corneal endothelium. The examined corneas included two cases of keratoconus complicated with acute hydrops and one without acute hydrops, two cases of herpetic keratitis, one case of post-intracapsular cataract extraction (post-ICCE) and one case of luetic keratitis. The endothelial cell layer was separated from Descemet''s membrane and double-stained with Rhodamine-labeled wheat germ agglutinin-lectin (WGA) and 4'',6-diamidino-2-phenylindole dihydrochloride (DAPI). The area of each cell was measured with a color image analyser and compared with its cytofluorometric nuclear DNA content. The endothelium in apparently intact regions of the diseased corneas showed the same DNA-ploidy pattern and cell area as the physiological corneas. However, endothelial cells in injured regions had greater area, even in diploidy, than in presumably normal ones and showed a larger number of hyperploid cells ranging from 4C to 36C. Hyperploid cells consisted of many multinucleates and few polyploidies and had extremely large and bizarre cytoplasm. All injured corneas were accompanied by cells with numerous micronuclei. A few asymmetrical 4C-binucleates (with DNA values such as 1.3 plus 2.6C) appeared in the case of the post-ICCE. It is concluded that damage to human corneal endothelial cells in vivo results in cell enlargement with or without DNA synthesis. Those changes appear more severe in diseased corneas than in the situation of physiological aging which we have reported previously. In severe cases, micronuclei, polyploid cells and multinucleated giant cells are frequent, thereby suggesting a possible long-persistent metabolic impairment of the endothelium after severe damage to the cornea.