School of Biomedical, Biomolecular and Chemical Sciences, the University of Western Australia, Crawley, WA 6009, Australia.
World J Gastroenterol. 2013 Feb 7;19(5):673-81. doi: 10.3748/wjg.v19.i5.673.
To evaluate whether desferrioxamine decreases ischemia and perfusion injury aggravated by cold storage (CS) in a rat liver perfusion model.
Isolated rat livers were kept in CS in University of Wisconsin Solution for 20 h at 4 °C, then exposed to 25 min of warm ischemia (WI) at 37 °C followed by 2 h of warm perfusion (WP) at 37 °C with oxygenated (95% oxygen and 5% carbon dioxide) Krebs-Henseleit buffer. Desferrioxamine (DFO), an iron chelator, was added at different stages of storage, ischemia and perfusion: in CS only, in WI only, in WP only, in WI and perfusion, or in all stages. Effluent samples were collected after CS and after WI. Perfusate samples and bile were collected every 30 min (0, 0.5, 1, 1.5 and 2 h) during liver perfusion. Cellular injury was assessed by the determination of lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) in the effluent and perfusate samples. Total iron was analysed in the perfusate samples. After WP, the liver was collected for the determination of liver swelling (wet to dry ratio) and liver morphological examination (hematoxylin and eosin staining).
Increased CS time caused increased liver dysfunction during WP. After 2 h of WP, liver injury was indicated by increased release of AST (0.5 h CS: 9.4 ± 2.2 U/g liver vs 20 h CS: 45.9 ± 10.8 U/g liver, P < 0.05) and LDH (0.5 h CS: 59 ± 14 U/g liver vs 20 h CS: 297 ± 71 U/g liver, P < 0.05). There was an associated increase in iron release into the perfusate (0.5 h CS: 0.11 ± 0.03 μmoL/g liver vs 20 h CS: 0.58 ± 0.10 μmoL/g liver, P < 0.05) and reduction in bile flow (0.5 h CS: 194 ± 12 μL/g vs 20 h CS: 71 ± 8 μL/g liver, P < 0.05). When DFO was added during WI and WP following 20 h of CS, release of iron into the perfusate was decreased (DFO absent 0.58 ± 0.10 μmoL/g liver vs DFO present 0.31 ± 0.06 μmoL/g liver, P < 0.05), and liver function substantially improved with decreased release of AST (DFO absent 45.9 ± 10.8 U/g liver vs DFO present 8.1 ± 0.9 U/g liver, P < 0.05) and LDH (DFO absent 297 ± 71 U/g liver vs DFO present 56 ± 7 U/g liver, P < 0.05), and increased bile flow (DFO absent 71 ± 8 μL/g liver vs DFO present 237 ± 36 μL/g liver, P < 0.05). DFO was also shown to improve liver morphology after WP. Cellular injury (the release of LDH and AST) was significantly reduced with the addition of DFO in CS medium but to a lesser extent compared to the addition of DFO in WP or WI and perfusion. There was no effect on liver swelling or bile flow when DFO was only added to the CS medium.
DFO added during WI and perfusion decreased liver perfusion injury aggravated by extended CS.
评估去铁胺是否会降低在大鼠肝脏灌注模型中冷储存（CS）加剧的缺血和灌注损伤。
将分离的大鼠肝脏在 4°C 的威斯康星大学溶液中 CS 保存 20 小时，然后在 37°C 下进行 25 分钟的热缺血（WI），然后在 37°C 下用含氧（95%氧气和 5%二氧化碳）的 Krebs-Henseleit 缓冲液进行 2 小时的热灌注（WP）。去铁胺（DFO），一种铁螯合剂，在储存、缺血和灌注的不同阶段添加：仅在 CS 中、仅在 WI 中、仅在 WP 中、在 WI 和灌注中或在所有阶段添加。在 CS 后和 WI 后收集流出物样本。在肝灌注过程中，每隔 30 分钟（0、0.5、1、1.5 和 2 小时）收集灌注液样本和胆汁。通过测定流出物和灌注液样本中的乳酸脱氢酶（LDH）和天冬氨酸氨基转移酶（AST）来评估细胞损伤。用灌注液样本中的总铁进行分析。WP 后，收集肝脏用于测定肝肿胀（湿重比）和肝形态学检查（苏木精和伊红染色）。
随着 WP 时间的延长，CS 时间的增加导致 WP 期间肝功能障碍增加。在 WP 2 小时后，AST（0.5 h CS：9.4±2.2 U/g 肝脏 vs 20 h CS：45.9±10.8 U/g 肝脏，P<0.05）和 LDH（0.5 h CS：59±14 U/g 肝脏 vs 20 h CS：297±71 U/g 肝脏，P<0.05）的释放表明肝损伤增加。铁释放到灌注液中也有相应增加（0.5 h CS：0.11±0.03 μmoL/g 肝脏 vs 20 h CS：0.58±0.10 μmoL/g 肝脏，P<0.05），胆汁流量减少（0.5 h CS：194±12 μL/g 与 20 h CS：71±8 μL/g 肝脏，P<0.05）。当在 20 小时 CS 后 WI 和 WP 期间添加 DFO 时，灌注液中铁的释放减少（DFO 不存在 0.58±0.10 μmoL/g 肝脏与 DFO 存在 0.31±0.06 μmoL/g 肝脏，P<0.05），AST（DFO 不存在 45.9±10.8 U/g 肝脏与 DFO 存在 8.1±0.9 U/g 肝脏，P<0.05）和 LDH（DFO 不存在 297±71 U/g 肝脏与 DFO 存在 56±7 U/g 肝脏，P<0.05）的释放显著改善，胆汁流量增加（DFO 不存在 71±8 μL/g 肝脏与 DFO 存在 237±36 μL/g 肝脏，P<0.05）。DFO 还显示在 WP 后改善肝形态。添加 DFO 可显著降低 CS 培养基中细胞损伤（LDH 和 AST 的释放），但与在 WP 或 WI 和灌注中添加 DFO 相比，其程度较小。当仅在 CS 培养基中添加 DFO 时，对肝肿胀或胆汁流量没有影响。
在 WI 和 WP 期间添加 DFO 可降低延长 CS 加剧的肝灌注损伤。
