Michael R, Vrensen G F, van Marle J, Gan L, Söderberg P G
Karolinska Institutet, St. Erik's Eye Hospital, Stockholm, Sweden.
Invest Ophthalmol Vis Sci. 1998 Dec;39(13):2681-7.
To investigate DNA damage in the rat lens after in vivo close-to-threshold exposure to ultraviolet radiation (UVR).
Sprague-Dawley rats received 5 kJ/m2 UVR (lambdaMAX = 300 nm, lambda0.5 = 10 nm) unilaterally for 15 minutes. Animals were killed at 1, 6, and 24 hours and at 1 week after exposure. DNA-strand breaks were investigated in sagittal paraffin sections using the TdT-dUTP terminal nick-end labeling (TUNEL) technique and propidium iodide for counterstaining. Other lenses were prepared for transmission electron microscopy (TEM).
TUNEL-positive nuclei were found at only 24 hours after UVR exposure. About one tenth of the epithelial cell nuclei were TUNEL positive, and affected cells were scattered over the entire epithelium. No TUNEL-positive cells were found at 1 or 6 hours or at 1 week after UVR exposure or in the nonexposed lenses. TEM verified the occurrence of programmed cell death and showed the breakdown of the apoptotic cells by adjacent cells. No signs of necrosis were found.
Threshold-dose UVR induces programmed cell death that peaks 24 hours after exposure and involves the entire epithelium. Dead cells are removed from the epithelium by phagocytosis.
研究大鼠晶状体在体内接近阈值剂量暴露于紫外线辐射(UVR)后的DNA损伤情况。
将Sprague-Dawley大鼠单侧暴露于5 kJ/m²的UVR(λMAX = 300 nm,λ0.5 = 10 nm)下15分钟。在暴露后1小时、6小时、24小时及1周处死动物。使用TdT-dUTP末端脱氧核苷酸转移酶介导的缺口末端标记(TUNEL)技术和碘化丙啶复染,在矢状石蜡切片中研究DNA链断裂情况。制备其他晶状体用于透射电子显微镜(TEM)观察。
仅在UVR暴露后24小时发现TUNEL阳性细胞核。约十分之一的上皮细胞核TUNEL阳性,受影响的细胞散布于整个上皮。在UVR暴露后1小时、6小时、1周或未暴露的晶状体中未发现TUNEL阳性细胞。TEM证实了程序性细胞死亡的发生,并显示凋亡细胞被相邻细胞分解。未发现坏死迹象。
阈值剂量的UVR诱导程序性细胞死亡,在暴露后24小时达到峰值,且累及整个上皮。死亡细胞通过吞噬作用从上皮中清除。