Nishiyama Y, Hayashi N, Murayama K, Mori K, Yoneya S
Department of Ophthalmalogy, Saitama Medical School, Japan.
Nippon Ganka Gakkai Zasshi. 1998 Jul;102(7):417-24.
50 eyes of 30 Sprague-Dawley rats were subjected to 60 minutes of pressure-induced ischemia, then fixed for light and electron microscopy with no reperfusion, or reperfusion after 30 minutes, 1, 2 or 4 hours, and 1 or 3 days from the time ocular ischemia was relaxed. The TdT-mediated dUTP-biotin nick end labeling (TUNEL) method revealed apoptotic signs at the inner retina as early as 1 hour after reperfusion. However, the incidence of apoptotic signs with the TUNEL method did not accord with the results of electron microscopic examination. During the time after the reperfusion started, especially after more than 4 hours, apoptotic signs became obvious and extended from the inner to the outer retina. These apoptotic findings could be seen with both the TUNEL method and electron microscopy. By 3 days after the reperfusion, necrotic cells in the ganglion cell layer, and the inner and outer nuclear layer became more prominent than apoptotic cells. These results may provide a baseline for therapeutic strategy and the prognosis of ischemia-reperfusion injury in the retina.
对30只Sprague-Dawley大鼠的50只眼睛施加60分钟的压力诱导性缺血,然后在不进行再灌注的情况下固定用于光镜和电镜检查,或者在眼缺血缓解后30分钟、1小时、2小时或4小时以及1天或3天进行再灌注后固定。TdT介导的dUTP生物素缺口末端标记(TUNEL)法显示,再灌注后1小时内视网膜内层即出现凋亡迹象。然而,TUNEL法检测到的凋亡迹象发生率与电镜检查结果不一致。在再灌注开始后的一段时间内,尤其是4小时后,凋亡迹象变得明显,并从视网膜内层扩展到外层。TUNEL法和电镜检查均能观察到这些凋亡表现。再灌注3天后,神经节细胞层以及内核层和外核层中的坏死细胞比凋亡细胞更为突出。这些结果可能为视网膜缺血再灌注损伤的治疗策略和预后提供基线。