Sasaki H, Matsuno T, Nakagawa K, Tanaka N
First Department of Surgery, Okayama University Medical School, Japan.
Acta Med Okayama. 1997 Dec;51(6):305-12. doi: 10.18926/AMO/30771.
This study was designed to investigate the induction of apoptosis during the reperfusion phase following warm liver ischemia in vivo. We evaluated apoptotic bodies (ABs) in sections stained with hematoxylin and eosin (H. E.) and positive hepatocytes in sections stained by the in situ nick end labeling method (TUNEL method) during the reperfusion phase up to 48 h after a 70% liver ischemia for 30 or 60 min in duration (30 or 60 min group). The peak number of ABs in H. E.-stained sections was observed at 1 to 3 h in the 30 min group and 3 to 6 h in the 60 min group. The number of ABs gradually fell as the length of the perfusion period increased, and few ABs were observed at 24 and 48 h after reperfusion. A peak number of TUNEL-positive hepatocytes was recognized at 3 h after reperfusion in both groups, after which the numbers decreased gradually. DNA extracted from both groups was electrophoresed on a 1.5% agarose gel. In both groups, a ladder-like pattern over smear pattern was recognized at 3 h after reperfusion. These results show that hepatocyte apoptosis was induced during the early phase of reperfusion after rat liver ischemia morphologically and biochemically, which suggests that hepatocyte apoptosis may be associated with ischemia and reperfusion injury.
本研究旨在探讨体内温性肝缺血后再灌注阶段细胞凋亡的诱导情况。我们评估了在70%肝缺血30或60分钟（30或60分钟组）后长达48小时的再灌注阶段，苏木精-伊红（H.E.）染色切片中的凋亡小体（ABs）以及原位缺口末端标记法（TUNEL法）染色切片中的阳性肝细胞。在H.E.染色切片中，30分钟组在1至3小时观察到ABs数量峰值，60分钟组在3至6小时观察到峰值。随着灌注时间延长，ABs数量逐渐下降，再灌注24和48小时后观察到的ABs很少。两组在再灌注3小时均识别出TUNEL阳性肝细胞数量峰值，之后数量逐渐减少。从两组提取的DNA在1.5%琼脂糖凝胶上进行电泳。两组在再灌注3小时均识别出在涂片模式上的梯状条带模式。这些结果表明，大鼠肝缺血后再灌注早期在形态学和生物化学上诱导了肝细胞凋亡，这表明肝细胞凋亡可能与缺血和再灌注损伤有关。
