Mitchell S J, Churchill T A, Winslet M C, Fuller B J
University Department of Surgery, Royal Free Hospital and School of Medicine, London, United Kingdom.
Cryobiology. 1996 Aug;33(4):413-22. doi: 10.1006/cryo.1996.0041.
Rat livers were flushed with different preservation solutions and stored at 4 degrees C for 24 h before being reperfused with a synthetic air-equilibrated, water-based solution. Four solutions were tested using this isolated rat liver model: Marshall's hypertonic citrate (HC); modified University of Wisconsin solution (Mod UW); a histidine-based solution (HIS); and a histidine-lactobionate-raffinose-based solution (HLR). After storage, livers were perfused at 4 degrees C for a period of 2 h and biopsy samples taken, at different time points, to investigate energy metabolism. Livers stored in HLR and HIS had higher 24-h storage levels of ATP (0.41 and 0.24 mumol/g respectively; P < 0.05) than those stored in Mod UW and HC. On reperfusion, all groups regenerated ATP by 1-2 h. However, significantly greater levels of ATP regeneration occurred in livers stored in the HLR (1.6 +/- 0.08 mumol/g) and Mod UW (1.3 +/- 0.18) than HC (0.58 +/- 0.19) and HIS (0.96 +/- 0.12); P < 0.05. Energy charge (1) (EC) recovered in all groups but was significantly higher in HLR and Mod UW (0.79 and 0.68, respectively; P < 0.05) than HC and HIS. These represent 95% (HLR) and 80% (Mod UW) of values observed in FIL. During the reperfusion period, total adenine nucleotide levels (TA) did not vary significantly within each storage group, except in the HIS solution. However, TAs were greater with livers stored for 24 h in HLR (2.5 +/- 0.25) and Mod UW (2.7 +/- 0.20) than those the other two storage groups (P < 0.05 in each case). This study has demonstrated that it was possible to resuscitate liver energetics after prolonged hypothermic ischemia by a period of cold reperfusion, and the method can differentiate between preservation solutions. The livers stored in each solution showed varying degrees of success in regeneration of ATP and EC, demonstrating that oxygen was not a limiting factor when using an air-equilibrated perfusate. The solutions providing the better preservation conditions gave the greater resuscitation of liver energetics (Mod UW and HLR). Overall, livers stored in HLR had the greatest resuscitation of energy metabolism, which correlates with survival data from other studies (29-31, 33).
用不同的保存溶液冲洗大鼠肝脏，并在4℃下储存24小时，然后用合成的空气平衡的水性溶液进行再灌注。使用这种离体大鼠肝脏模型测试了四种溶液：马歇尔高渗柠檬酸盐（HC）；改良的威斯康星大学溶液（Mod UW）；一种基于组氨酸的溶液（HIS）；以及一种基于组氨酸-乳糖酸-棉子糖的溶液（HLR）。储存后，肝脏在4℃下灌注2小时，并在不同时间点采集活检样本以研究能量代谢。储存在HLR和HIS中的肝脏在24小时储存时的ATP水平（分别为0.41和0.24μmol/g；P<0.05）高于储存在Mod UW和HC中的肝脏。再灌注时，所有组在1-2小时内再生ATP。然而，储存在HLR（1.6±0.08μmol/g）和Mod UW（1.3±0.18）中的肝脏的ATP再生水平明显高于HC（0.58±0.19）和HIS（0.96±0.12）；P<0.05。所有组的能量电荷（1）（EC）均恢复，但HLR和Mod UW中的能量电荷明显更高（分别为0.79和0.68；P<0.05），高于HC和HIS。这些分别代表在FIL中观察到的值的95%（HLR）和80%（Mod UW）。在再灌注期间，除了在HIS溶液中，每个储存组内的总腺嘌呤核苷酸水平（TA）没有显著变化。然而，在HLR（2.5±0.25）和Mod UW（2.7±0.20）中储存24小时的肝脏的TA高于其他两个储存组（每种情况下P<0.05）。这项研究表明，通过一段时间的冷再灌注，在长时间低温缺血后恢复肝脏能量学是可能的，并且该方法可以区分保存溶液。储存在每种溶液中的肝脏在ATP和EC再生方面显示出不同程度的成功，表明当使用空气平衡的灌注液时，氧气不是限制因素。提供更好保存条件的溶液能使肝脏能量学得到更大程度的恢复（Mod UW和HLR）。总体而言，储存在HLR中的肝脏在能量代谢恢复方面最为显著，这与其他研究（29-31,33）的存活数据相关。
