Borderie V M, Baudrimont M, Vallée A, Ereau T L, Gray F, Laroche L
Department of Ophthalmology, Hôpital Saint Antoine, Paris, France.
Invest Ophthalmol Vis Sci. 2000 Aug;41(9):2501-5.
To investigate whether apoptosis plays a notable role in degeneration of corneal endothelial cells in patients with Fuchs' dystrophy.
Forty-seven corneal buttons from 41 patients with Fuchs' dystrophy were studied. Nucleus labeling, transmission electron microscopy (TEM), and TdT-dUTP terminal nick-end labeling (TUNEL) were used to detect apoptosis. TEM and TUNEL were performed on sections of all 47 corneal buttons, and nucleus labeling was performed on the last 10 corneas. Seven human donor corneas, two corneal buttons from two patients with keratoconus, and one corneal button from a patient with interstitial keratitis were used as negative controls for detection of apoptotic endothelial cells. Negative controls were studied by means of nucleus labeling, TUNEL, and TEM.
In the nucleus labeling assay, the average percentage of apoptotic endothelial cells was 2.65% in the Fuchs' dystrophy group (n = 10) and 0.23% in the control group (n = 10; P = 0.0003). In the TUNEL assay, labeling of some endothelial cells was observed on 42 of 47 corneas in the Fuchs' dystrophy group, whereas it was absent on most specimens of the control group. In TEM, most endothelial cell nuclei had a normal appearance, and apoptotic endothelial cells featuring condensed nucleus and decreased cell size could be observed exceptionally. Some apoptotic cells were found in the basal epithelial cell layer by means of nucleus labeling, TUNEL, and TEM in the Fuchs' dystrophy group but not in the control group.
This study suggests that apoptosis plays an important role in endothelial cell degeneration in Fuchs' dystrophy. Because of a lack of conclusive evidence of increased endothelial apoptosis by TEM, further studies are needed to ascertain this finding.
研究细胞凋亡在富克斯角膜内皮营养不良患者角膜内皮细胞退变中是否起显著作用。
对41例富克斯角膜内皮营养不良患者的47个角膜植片进行研究。采用细胞核标记、透射电子显微镜(TEM)和TdT-dUTP末端脱氧核苷酸转移酶介导的缺口末端标记法(TUNEL)检测细胞凋亡。对所有47个角膜植片进行TEM和TUNEL检测,对最后10个角膜进行细胞核标记。7个供体角膜、2例圆锥角膜患者的2个角膜植片和1例间质性角膜炎患者的1个角膜植片用作检测凋亡内皮细胞的阴性对照。通过细胞核标记、TUNEL和TEM对阴性对照进行研究。
在细胞核标记检测中富克斯角膜内皮营养不良组(n = 10)凋亡内皮细胞的平均百分比为2.65%,对照组(n = 10;P = 0.0003)为0.23%。在TUNEL检测中,富克斯角膜内皮营养不良组47个角膜中的42个观察到一些内皮细胞有标记,而对照组大多数标本未观察到标记。在TEM检测中,大多数内皮细胞核外观正常,仅偶尔能观察到核浓缩和细胞大小减小的凋亡内皮细胞。在富克斯角膜内皮营养不良组,通过细胞核标记、TUNEL和TEM在基底上皮细胞层发现了一些凋亡细胞,而对照组未发现。
本研究提示细胞凋亡在富克斯角膜内皮营养不良内皮细胞退变中起重要作用。由于TEM缺乏内皮细胞凋亡增加的确凿证据,需要进一步研究以确定这一发现。