Apoptosis in UV-exposed Rabbit Corneas
Apoptosis in UV-exposed Rabbit Corneas
复制标题
紫外线暴露的兔角膜细胞凋亡
作者：
A. Podskochy;L. Gan;P. Fagerholm
目的.本实验研究了不同波长紫外线照射引起的兔角膜光性角膜炎后细胞凋亡的可能机制。法将14只白化病兔角膜暴露于10 nm全波段的280 nm和310 nm紫外线辐射（UVR），其剂量可引起生物显微镜下显著的角膜炎（280 nm为0.12 J/cm 2，310 nm为0.47 J/cm 2）。暴露后24和76 h处死动物。对角膜进行光镜和透射电子显微镜检查，并使用修改的TUNEL技术对片段化DNA进行原位末端标记。结果角膜暴露于280 nm的紫外线照射后24和76小时，TUNEL阳性染色仅在上皮细胞和浅层角膜细胞。310 nm紫外线照射24小时后，上皮细胞、中央基质整个厚度的角膜基质细胞和内皮细胞中出现TUNEL阳性染色。暴露于310-nm UVR后76小时，整个受损基质层的角膜基质细胞消失。只有少数上皮细胞TUNEL阳性。透射电子显微镜（TEM）证实了凋亡细胞核和细胞的发生。结论细胞凋亡似乎是UVR后角膜细胞死亡的一种机制。310-nm UVR对角膜基质和内皮的损伤比280-nm UVR更广泛。
Purpose. Apoptosis was studied in rabbit corneas as a possible mechanism of cell death after photokeratitis induced by different UV wavelengths. Method. Fourteen albino rabbit corneas were exposed to 280-and 310-nm UV radiation (UVR) in 10-nm full wavebands at doses that cause biomicroscopically significant keratitis (0.12 J/cm2 for 280 nm and 0.47 J/cm2 for 310 nm). Animals were killed 24 and 76 h after exposure. Corneas were processed for light and transmission electron microscopy and in situ end labeling of fragmented DNA by using a modification of the TUNEL technique. Results. Corneas exposed to 280-nm UVR showed TUNEL-positive staining only in epithelial cells and superficial keratocytes at 24 and 76 h after irradiation. Twenty-four hours after 310-nm UVR exposure, TUNEL-positive staining was present in the epithelial cells, keratocytes throughout the entire thickness of the central stroma, and in endothelial cells. Seventy-six hours after exposure to 310-nm UVR, keratocytes disappeared throughout the whole thickness of the damaged stroma. Only a few epithelial cells were TUNEL positive at that time. Transmission electron microscopy (TEM) verified the occurrence of apoptotic nuclei and cells. Conclusion. Apoptosis appears to be a mechanism of corneal cell death after UVR. The 310-nm UVR caused more extensive damage to the corneal stroma and endothelium than did the 280-nm UVR.
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