Cook B, Lewis G P, Fisher S K, Adler R
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Invest Ophthalmol Vis Sci. 1995 May;36(6):990-6.
To investigate the possibility that cell death in retinal detachment may occur by reactivation of apoptotic programmed cell death mechanisms.
Unilateral retinal detachments were created in adult cats using 0.25% sodium hyaluronate; detached and control retinas were studied at different intervals. Internucleosomal DNA fragmentation (one of the landmarks of apoptosis) was investigated in tissue sections with the TUNEL technique, which uses terminal transferase to label with biotinylated nucleotides the 3' ends of DNA fragments. Sections also were labeled with propidium iodide, which intensely stains pyknotic nuclei. In addition, one time point was selected for analysis with electron microscopy.
TUNEL-positive (T+) and propidium iodide-positive (PI+) cells almost never were observed in retinas from control eyes, but they were abundant at defined time points after retinal detachment, appearing almost exclusively in the photoreceptor layer. Their frequency was particularly high 1 to 3 days after detachment but declined rapidly over the next several weeks. T+ cells were still present 28 days after retinal detachment. Electron microscopy also revealed evidence of apoptotic cells after retinal detachment.
Results are consistent with the hypothesis that photoreceptor degeneration after retinal detachment occurs through apoptosis, usually associated with intrinsic, programmed cell death mechanisms. The detection of a rapid wave of photoreceptor degeneration seems to suggest that early therapeutic interventions might be recommended; agents capable of interfering with the apoptotic mechanism could have a role in the prevention of cell losses that represent a critical complication of retinal detachment.
研究视网膜脱离时细胞死亡是否可能通过凋亡性程序性细胞死亡机制的重新激活而发生。
使用0.25%透明质酸钠在成年猫中造成单侧视网膜脱离;在不同时间间隔对脱离的视网膜和对照视网膜进行研究。采用TUNEL技术在组织切片中研究核小体间DNA片段化(凋亡的标志之一),该技术利用末端转移酶用生物素化核苷酸标记DNA片段的3'末端。切片还用碘化丙啶标记,碘化丙啶可强烈染色固缩核。此外,选择一个时间点进行电子显微镜分析。
在对照眼的视网膜中几乎从未观察到TUNEL阳性(T+)和碘化丙啶阳性(PI+)细胞,但在视网膜脱离后的特定时间点它们大量存在,几乎仅出现在光感受器层。它们的频率在脱离后1至3天特别高,但在接下来的几周内迅速下降。视网膜脱离28天后仍有T+细胞存在。电子显微镜也显示了视网膜脱离后凋亡细胞的证据。
结果与以下假设一致,即视网膜脱离后光感受器变性是通过凋亡发生的,通常与内在的程序性细胞死亡机制有关。快速的光感受器变性浪潮的检测似乎表明可能推荐早期治疗干预;能够干扰凋亡机制的药物可能在预防代表视网膜脱离关键并发症的细胞损失中发挥作用。