Caraceni P, Gasbarrini A, Nussler A, Di Silvio M, Bartoli F, Borle A B, Van Thiel D H
Oklahoma Transplant Institute, Baptist Medical Center of Oklahoma, Oklahoma City 73112.
Hepatology. 1994 Nov;20(5):1247-54.
We performed this study to determine whether perfused isolated human and rat hepatocytes have different sensitivities to anoxia-reoxygenation injury. Oxygen free radicals were detected by lucigenin-enhanced chemiluminescence. Lipid peroxidation was assessed by measuring malondialdehyde release. Cell injury was evaluated by measuring lactate dehydrogenase release and trypan blue uptake. During the control period, lucigenin-enhanced chemiluminescence, malondialdehyde and lactate dehydrogenase release and trypan blue uptake were similar in rat and human hepatocytes. During 3.5 hr of anoxia, lucigenin-enhanced chemiluminescence decreased to background levels and malondialdehyde release remained constant in both groups. In contrast, lactate dehydrogenase release increased eightfold in rat hepatocytes but only threefold in human hepatocytes. With reoxygenation after 2.5 hr of anoxia, in rat hepatocytes lucigenin-enhanced chemiluminescence increased 13-fold within 15 min and then declined toward control levels. Malondialdehyde release doubled after 1 hr of reoxygenation. The rate of lactate dehydrogenase release increased to a level almost twice that observed in cells kept continuously anoxic. In contrast, with human hepatocytes lucigenin-enhanced chemiluminescence increased only fourfold, whereas malondialdehyde and lactate dehydrogenase releases did not differ significantly from those levels measured in cells perfused continuously under anoxic conditions. At the end of the experiment, the increase in trypan blue uptake was significantly greater with rat hepatocytes than with human hepatocytes. These results demonstrate that (a) during reoxygenation following 2.5 hr of anoxia, isolated human hepatocytes generate fewer oxygen free radical, and lipoperoxides than do rat hepatocytes, and (b) human hepatocytes are more resistant to cell injury during anoxia-reoxygenation than are rat hepatocytes.
我们开展这项研究以确定灌注的分离人肝细胞和大鼠肝细胞对缺氧-复氧损伤是否具有不同的敏感性。通过光泽精增强化学发光法检测氧自由基。通过测量丙二醛释放量评估脂质过氧化。通过测量乳酸脱氢酶释放量和台盼蓝摄取量评估细胞损伤。在对照期，大鼠和人肝细胞的光泽精增强化学发光、丙二醛和乳酸脱氢酶释放量以及台盼蓝摄取量相似。在缺氧3.5小时期间，两组的光泽精增强化学发光均降至背景水平，丙二醛释放量保持恒定。相比之下，大鼠肝细胞的乳酸脱氢酶释放量增加了8倍，而人肝细胞仅增加了3倍。缺氧2.5小时后复氧，大鼠肝细胞的光泽精增强化学发光在15分钟内增加了13倍，然后降至对照水平。复氧1小时后丙二醛释放量增加了一倍。乳酸脱氢酶释放速率增加到几乎是持续缺氧细胞中观察到水平的两倍。相比之下，人肝细胞的光泽精增强化学发光仅增加了4倍，而丙二醛和乳酸脱氢酶释放量与在缺氧条件下持续灌注的细胞中测得的水平相比无显著差异。在实验结束时，大鼠肝细胞的台盼蓝摄取增加量显著高于人肝细胞。这些结果表明：(a) 在缺氧2.5小时后的复氧过程中，分离的人肝细胞产生的氧自由基和脂质过氧化物比大鼠肝细胞少；(b) 在缺氧-复氧过程中，人肝细胞比大鼠肝细胞对细胞损伤更具抗性。
