Boudjema K, Lindell S L, Belzer F O, Southard J H
Department of Surgery, University of Wisconsin, Madison 53792.
Cryobiology. 1991 Jun;28(3):227-36. doi: 10.1016/0011-2240(91)90027-l.
Livers from fed, fasted (48 h) and glucose-fed rabbits were preserved for 24 and 48 h by either simple cold storage (CS) or continuous machine perfusion (MP) with the University of Wisconsin preservation solutions. After preservation liver functions were measured by isolated perfusion of the liver (at 37 degrees C) for 2 h. Fasting caused an 85% reduction in the concentration of glycogen in the liver but no change in ATP or glutathione. Glucose feeding suppressed the loss of glycogen (39% loss). After 24 h preservation by CS livers from fed or fasted animals were similar including bile production (6.2 +/- 0.5 and 5.6 +/- 0.4 ml/2 h, 100 g, respectively), hepatocellular injury (LDH release = 965 +/- 100 and 1049 +/- 284 U/liter), and concentrations of ATP (1.17 +/- 0.15 and 1.18 +/- 0.04 mumol/g, glutathione (1.94 +/- 0.51 and 2.35 +/- 0.26 mumol/g, respectively), and K:Na ratio (6.7 +/- 1.0 and 7.7 +/- 0.5, respectively). After 48 h CS livers from fed animals were superior to livers from fasted animals including significantly more bile production (5.0 +/- 0.9 vs 2.0 +/- 0.3 ml/2 h, 100 g), less LDH release (1123 +/- 98 vs 3701 +/- 562 U/liter), higher concentration of ATP (0.50 +/- 0.16 vs 0.33 +/- 0.07 mumol/g) and glutathione (0.93 +/- 0.14 vs 0.30 +/- 0.13 mumol/g), and a larger K:Na ratio (7.4 vs 1.5). Livers from fed animals were also better preserved than livers from fasted animals when the method was machine perfusion. The decrease in liver functions in livers from fasted animals preserved for 48 h by CS or MP was prevented by feeding glucose. Glucose feeding increased bile formation after 48 h CS preservation from 2.0 +/- 0.3 (fasted) to 6.9 +/- 1.2 ml/2 h, 100 g; LDH release was reduced from 3701 +/- 562 (fasted) to 1450 +/- 154 U/liter; ATP was increased from 0.33 +/- 0.07 (fasted) to 1.63 +/- 0.18 mumol/g; glutathione was increased from 0.30 +/- 0.01 (fasted) to 2.17 +/- 0.30 mumol g; and K:Na ratio was increased from 1.5 +/- 0.9 to 5.3 +/- 1.0. This study shows that the nutritional status of the donor can affect the quality of liver preservation. The improvement in preservation by feeding rabbits only glucose suggests that glycogen is an important metabolite for successful liver preservation. Glycogen may be a source for ATP synthesis during the early period of reperfusion of preserved livers.
将喂食、禁食(48小时)和葡萄糖喂食的兔子的肝脏,通过单纯冷藏(CS)或用威斯康星大学保存液进行连续机器灌注(MP)的方式保存24小时和48小时。保存后,通过在37摄氏度下对肝脏进行2小时的离体灌注来测量肝功能。禁食导致肝脏中糖原浓度降低85%,但三磷酸腺苷(ATP)或谷胱甘肽没有变化。葡萄糖喂食抑制了糖原的损失(损失39%)。通过CS保存24小时后,喂食或禁食动物的肝脏相似,包括胆汁生成量(分别为6.2±0.5和5.6±0.4毫升/2小时,100克)、肝细胞损伤(乳酸脱氢酶(LDH)释放量=965±100和1049±284单位/升),以及ATP浓度(1.17±0.15和1.18±0.04微摩尔/克)、谷胱甘肽(1.94±0.51和2.35±0.26微摩尔/克),以及钾钠比(分别为6.7±1.0和7.7±0.5)。通过CS保存48小时后,喂食动物的肝脏优于禁食动物的肝脏,包括胆汁生成量显著更多(5.0±0.9对2.0±0.3毫升/2小时,100克)、LDH释放量更少(1123±98对3701±562单位/升)、ATP浓度更高(0.50±0.16对0.33±0.07微摩尔/克)和谷胱甘肽(0.93±0.14对0.30±0.13微摩尔/克),以及更大的钾钠比(7.4对1.5)。当采用机器灌注方法时,喂食动物的肝脏也比禁食动物的肝脏保存得更好。通过CS或MP保存48小时的禁食动物肝脏的肝功能下降,通过喂食葡萄糖得以预防。在CS保存48小时后,葡萄糖喂食使胆汁生成量从2.0±0.3(禁食)增加到6.9±1.2毫升/2小时,100克;LDH释放量从3701±562(禁食)降低到1450±154单位/升;ATP从0.33±0.07(禁食)增加到1.63±0.18微摩尔/克;谷胱甘肽从0.30±0.01(禁食)增加到2.17±0.30微摩尔/克;钾钠比从1.5±0.9增加到5.3±1.0。本研究表明,供体的营养状况会影响肝脏保存的质量。仅通过喂食兔子葡萄糖来改善保存效果表明,糖原是成功进行肝脏保存的重要代谢物。糖原可能是保存肝脏再灌注早期ATP合成的来源。