Podskochy A, Gan L, Fagerholm P
St. Erik's Eye Hospital, Karolinska Institutet, Stockholm, Sweden.
Cornea. 2000 Jan;19(1):99-103. doi: 10.1097/00003226-200001000-00019.
Apoptosis was studied in rabbit corneas as a possible mechanism of cell death after photokeratitis induced by different UV wavelengths.
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.
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.
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.
研究兔角膜中的细胞凋亡,作为不同紫外线波长诱导光性角膜炎后细胞死亡的一种可能机制。
14只白化兔角膜分别暴露于280纳米和310纳米的紫外线辐射(UVR),以10纳米的全波段,剂量设定为引起生物显微镜下显著角膜炎的剂量(280纳米为0.12焦耳/平方厘米,310纳米为0.47焦耳/平方厘米)。动物在暴露后24小时和76小时处死。角膜进行光镜和透射电镜处理,并使用改良的TUNEL技术对断裂DNA进行原位末端标记。
暴露于280纳米UVR的角膜在照射后24小时和76小时仅在上皮细胞和浅层角膜细胞中显示TUNEL阳性染色。在310纳米UVR暴露后24小时,上皮细胞、中央基质全层的角膜细胞以及内皮细胞中均存在TUNEL阳性染色。在暴露于310纳米UVR 76小时后,受损基质全层的角膜细胞消失。此时只有少数上皮细胞TUNEL阳性。透射电子显微镜(TEM)证实了凋亡细胞核和细胞的存在。
细胞凋亡似乎是UVR后角膜细胞死亡的一种机制。310纳米的UVR比280纳米的UVR对角膜基质和内皮造成的损伤更广泛。