新型SMO多器官保存液在大鼠肝脏保存中的实验研究
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摘要
第一章新型SMO保存液中聚乙二醇分子量及浓度的选择
目的:探讨新型SMO保存液中聚乙二醇的最佳分子量和浓度。
材料和方法:大鼠肝脏原位灌注切取后在不同SMO保存液中4℃保存24h,保存终点行肝脏组织病理学检查和评分,比较含不同分子量、不同浓度聚乙二醇(PEG)的SMO保存液对大鼠肝脏冷保存的效果。将人静脉血与含不同分子量、不同浓度PEG的SMO保存液混合,通过自动血沉分析仪检测红细胞沉降率、自动血液流变仪检测血液流变学指标、光镜观察红细胞聚集的形态学改变,分析PEG对红细胞聚集性和血液流变学的影响。
结果:在相同浓度下,含PEG 35KDa(PEG35)的SMO保存液对大鼠肝脏的低温保存效果优于含PEG 20KDa(PEG20)的保存液,其中PEG35(1g/L)的保存效果最好。含低浓度PEG的SMO保存液对红细胞聚集性和血液流变学的影响较小;而含PEG20(30g/L)、PEG35(10g/L)和PEG35(30g/L)的保存液则可显著加快红细胞沉降率,增加血液粘滞度,引起红细胞明显聚集。
结论:PEG35(1g/L)是新型SMO保存液中PEG的最佳选择。
第二章大鼠肝脏离体再灌注模型的建立及改进
目的:建立大鼠肝脏离体再灌注模型,为器官保存液的研究提供可靠平台。
材料和方法:在国外同类系统的基础上,对其进行改进,建立适合我国国情的长征-1(CZ-1)型大鼠肝脏离体再灌注系统。随后对系统的有效性和可靠性进行了实验验证。大鼠肝脏经门静脉、胆道分别插管后切取,通过CZ-1型离体再灌注系统在37℃条件下以20ml/min的流量经门静脉循环灌注Krebs-Henseleit液120min。观察肝脏胆汁分泌量和组织病理学改变,探讨大鼠肝脏离体再灌注模型的有效性和可靠性。
结果:CZ-1型大鼠肝脏离体再灌注系统价格低廉、结构简单、性能可靠。离体再灌注肝脏的胆汁分泌量为0.248±0.094 ul/min·g(肝组织),与国外文献报道类似。再灌注肝脏组织病理学无明显改变。
结论:大鼠肝脏离体再灌注模型是器官保存液研究的理想模型。CZ-1型大鼠肝脏离体再灌注系统简单有效,可替代国外进口系统。
第三章新型SMO保存液在大鼠肝脏保存中的实验研究
目的:探讨新型SMO保存液对大鼠肝脏的保存效果。
材料和方法:大鼠肝脏原位灌注切取后,在UW液和SMO液中4℃分别保存24、48、72h,冷保存终点行肝脏组织病理学检查、保存液pH值和渗透压测定、肝脏增重检测,探讨SMO保存液的冷保存效果。另一部分大鼠肝脏原位灌注切取后,在乳酸林格液、UW液和SMO液中4℃保存24h,未冷保存组为对照,冷保存后通过离体再灌注系统,在37℃条件下以20ml/min的流量经门静脉循环灌注Krebs-Henseleit液120min,通过生化检测观察灌注液中ALT、AST、LDH的改变,灌注结束后通过光镜、电镜观察肝脏组织病理学改变,通过TUNEL实验检测凋亡情况,通过免疫组化检测肝组织中NF-κB、ICAM-1、Caspase-3的表达,探讨SMO保存液在减轻肝脏冷缺血再灌注损伤中的作用。结果:冷保存24h后,SMO液组与UW液组的肝脏保存效果类似;冷保存48h后,
SMO液组的肝脏保存效果略差于UW液组,但无显著性差异。冷保存24、48、72h后,SMO液组的肝脏增重分别为5.53±4.21%、6.57±3.24%和8.22±2.96%,而UW液组的肝脏增重为-8.04±6.42%、-12.31±7.61%和-18.58±8.80%。冷保存24、48、72h后, SMO液组pH值分别为7.23±0.04、7.07±0.05、6.98±0.04,而UW液组pH值分别为7.12±0.04、6.93±0.05、6.82±0.04,但两者缓冲能力类似。随着保存时间的延长,SMO液组的渗透压缓慢升高,而UW液组的渗透压升高较快。在冷保存24h后,两组的酶学指标无明显性差异,但灌注120min后,SMO液组ALT值低于UW液组,而AST和LDH则高于UW液组。离体再灌注120min后,SMO液组的光镜组织病理学评分显著差于UW液组,而电镜检查结果两组无显著性差异。TUNEL结果显示,SMO组的肝细胞凋亡率高于UW液组,但无显著性差异。免疫组化显示,SMO组中NF-κB、ICAM-1、Caspase-3的阳性表达率与UW组类似,无显著性差异。
结论:SMO保存液对大鼠肝脏冷缺血损伤的保护效果与UW液类似,但其减轻冷缺血再灌注损伤的能力弱于UW液。
Chapter 1 The optimal molecular weight and concentration of PEG added to the newly developed SMO solution
Objective: To evaluate the optimal molecular weight and concentration of PEG added to the newly developed SMO solution.
Methods: Livers from SD rats were flushed and then preserved at 4°C for 24h in SMO solution with different molecular weight and concentration of PEG (no PEG, 1g/L PEG20, 10g/L PEG20, 30g/L PEG20, 1g/L PEG35, 10g/L PEG35 and 30g/L PEG35). At the end of the preservation, histological examination of hepatic tissues was performed. Then human venous blood was mixed with normal saline, UW solution, SMO solutions with different molecular weight and concentration of PEG. The dilution ratios of blood with preservation solution were 5:1 and 1:1. Sedimentation rate was measured by an automatic ESR analyzer. Human RBC aggregability and blood viscosity was investigated with an automatic hemorheological analyzer. Light microscopy was used to evaluate morphological characters of the RBC aggregates.
Results: After cold preservation for 24h, SMO solution with PEG35 (1g/L) showed the best preservation effect among the tested solution. PEG20 (1 and 10g/L) and PEG35 (1g/L) had little effect on RBC aggregation, while PEG20 (30g/L) and PEG35 (10 and 30g/L) significantly increased the RBC sedimentation and blood viscosity, and had a hyperaggregating effect on RBC.
Conclusion: Low concentration PEG35 (1g/L) would be the optimal choice added to the SMO solution.
Chapter 2 Establishment of the isolated perfused rat liver model and its modification
Objective: To set up the isolated perfused rat liver model, provide a reliable study platform for the development of organ preservation solution.
Methods: We modified the isolated perfused system described previously in references and establish CZ - 1 isolated perfused rat liver system. Then the effectiveness of the isolated perfused system was tested. Livers were harvested after the cannulation of the portal vein and bile duct. Then Livers were connected via the portal vein to a recirculating perfusion system for 120min. The reperfusion solution was Krebs-Henseleit solution at a constant temperature of 37°C and a flow rate of 20ml/min. After 120min reperfusion, bile production was evaluated. Routine HE staining examination of hepatic tissues was also performed.
Results: The CZ - 1 isolated perfused rat liver system was cost-effective and reliable to use. It was easy to run. The bile volume collected from the isolated repefused liver were 0.248±0.094 ul/min·g (liver) and was not significantly different from that reported by references. Hepatic tissues in reperfusion group were also morphologically normal.
Conclusion: The isolated perfused rat liver model closely mimics physiologic conditions and is the ideal model for investigation of organ preservation solution.
Chapter 3 Effects of SMO solution in rat liver preservation
Objective: To compare the protective effect of SMO solution with that of UW solution during cold preservation and normothermic reperfusion.
Methods: SD rats were divided into four groups according to different preservation solution: control group (without cold preservation), lactated Ringers group (negative control), UW group and SMO group. Preservation injury after cold storage for 24, 48 and 72h was assessed microscopically. Meanwhile the pH value and osmotic pressure of the preservation solutions was measured. The change of the liver weight was also assessed. After 24h cold storage of rat liver in different preservation solutions, the isolated perfused rat liver model was applied to reperfuse the liver for 120min normothermically with Krebs-Henseleit solution. Reperfusion injury was analyzed in the isolated perfused liver by enzyme (AST, ALT and LDH) release and bile production. Routine HE staining and electron microscopic examination of hepatic tissues were performed. NF-κB, ICAM-1 and Caspase-3 expressions were assessed by immunohistochemical analysis. Hepatocellular apoptosis was also assessed
Results: After preserving for 24h, histology showed no difference between the SMO and UW group. But after 48h cold preservation, UW solution provided rat livers with better protection against cold ischemia injury. During cold preservation, pH value of the SMO and UW solution decreased with preservation time, and the buffering capacity of both solutions was similar. The osmotic pressure of the both preservation solutions increased with preservation time, but they were higher in UW solution. Prolonged cold preaervation time resulted in an increase in liver weight with SMO-preserved livers and a decrease in weight with UW-preserved livers. After 24h cold storage, transaminase level in the both groups showed no difference. After reperfusion for 120min, ALT level was lower in SMO group, but higher AST and LDH levels were found in SMO group than in UW group. The amount of Bile product after reperfusion for 120min in UW group was more than that in SMO group. Morphological changes in SMO group were significiently worse than that in UW group. Histological results showed more necrotic regions in livers preserved in SMO solution. Electron microscopy revealed that ultrastructural impairments were similar in both groups. No significant differences were found in NF-κB and ICAM-1 expression between SMO group and UW group. The percentage of apoptotic cells and the expression patterns of apoptosis-related-genes were similar in livers preserved in SMO and UW solutions. The variables in both groups were better than those of livers preserved in Ringers solution.
Conclusion: The short-term cold preservation effect of SMO solution is similar with that of UW solution. However, SMO solution is inferior to UW solution in protecting the liver from cold ischemia-reperfusion injury.
目的:探讨新型SMO保存液中聚乙二醇的最佳分子量和浓度。
材料和方法:大鼠肝脏原位灌注切取后在不同SMO保存液中4℃保存24h,保存终点行肝脏组织病理学检查和评分,比较含不同分子量、不同浓度聚乙二醇(PEG)的SMO保存液对大鼠肝脏冷保存的效果。将人静脉血与含不同分子量、不同浓度PEG的SMO保存液混合,通过自动血沉分析仪检测红细胞沉降率、自动血液流变仪检测血液流变学指标、光镜观察红细胞聚集的形态学改变,分析PEG对红细胞聚集性和血液流变学的影响。
结果:在相同浓度下,含PEG 35KDa(PEG35)的SMO保存液对大鼠肝脏的低温保存效果优于含PEG 20KDa(PEG20)的保存液,其中PEG35(1g/L)的保存效果最好。含低浓度PEG的SMO保存液对红细胞聚集性和血液流变学的影响较小;而含PEG20(30g/L)、PEG35(10g/L)和PEG35(30g/L)的保存液则可显著加快红细胞沉降率,增加血液粘滞度,引起红细胞明显聚集。
结论:PEG35(1g/L)是新型SMO保存液中PEG的最佳选择。
第二章大鼠肝脏离体再灌注模型的建立及改进
目的:建立大鼠肝脏离体再灌注模型,为器官保存液的研究提供可靠平台。
材料和方法:在国外同类系统的基础上,对其进行改进,建立适合我国国情的长征-1(CZ-1)型大鼠肝脏离体再灌注系统。随后对系统的有效性和可靠性进行了实验验证。大鼠肝脏经门静脉、胆道分别插管后切取,通过CZ-1型离体再灌注系统在37℃条件下以20ml/min的流量经门静脉循环灌注Krebs-Henseleit液120min。观察肝脏胆汁分泌量和组织病理学改变,探讨大鼠肝脏离体再灌注模型的有效性和可靠性。
结果:CZ-1型大鼠肝脏离体再灌注系统价格低廉、结构简单、性能可靠。离体再灌注肝脏的胆汁分泌量为0.248±0.094 ul/min·g(肝组织),与国外文献报道类似。再灌注肝脏组织病理学无明显改变。
结论:大鼠肝脏离体再灌注模型是器官保存液研究的理想模型。CZ-1型大鼠肝脏离体再灌注系统简单有效,可替代国外进口系统。
第三章新型SMO保存液在大鼠肝脏保存中的实验研究
目的:探讨新型SMO保存液对大鼠肝脏的保存效果。
材料和方法:大鼠肝脏原位灌注切取后,在UW液和SMO液中4℃分别保存24、48、72h,冷保存终点行肝脏组织病理学检查、保存液pH值和渗透压测定、肝脏增重检测,探讨SMO保存液的冷保存效果。另一部分大鼠肝脏原位灌注切取后,在乳酸林格液、UW液和SMO液中4℃保存24h,未冷保存组为对照,冷保存后通过离体再灌注系统,在37℃条件下以20ml/min的流量经门静脉循环灌注Krebs-Henseleit液120min,通过生化检测观察灌注液中ALT、AST、LDH的改变,灌注结束后通过光镜、电镜观察肝脏组织病理学改变,通过TUNEL实验检测凋亡情况,通过免疫组化检测肝组织中NF-κB、ICAM-1、Caspase-3的表达,探讨SMO保存液在减轻肝脏冷缺血再灌注损伤中的作用。结果:冷保存24h后,SMO液组与UW液组的肝脏保存效果类似;冷保存48h后,
SMO液组的肝脏保存效果略差于UW液组,但无显著性差异。冷保存24、48、72h后,SMO液组的肝脏增重分别为5.53±4.21%、6.57±3.24%和8.22±2.96%,而UW液组的肝脏增重为-8.04±6.42%、-12.31±7.61%和-18.58±8.80%。冷保存24、48、72h后, SMO液组pH值分别为7.23±0.04、7.07±0.05、6.98±0.04,而UW液组pH值分别为7.12±0.04、6.93±0.05、6.82±0.04,但两者缓冲能力类似。随着保存时间的延长,SMO液组的渗透压缓慢升高,而UW液组的渗透压升高较快。在冷保存24h后,两组的酶学指标无明显性差异,但灌注120min后,SMO液组ALT值低于UW液组,而AST和LDH则高于UW液组。离体再灌注120min后,SMO液组的光镜组织病理学评分显著差于UW液组,而电镜检查结果两组无显著性差异。TUNEL结果显示,SMO组的肝细胞凋亡率高于UW液组,但无显著性差异。免疫组化显示,SMO组中NF-κB、ICAM-1、Caspase-3的阳性表达率与UW组类似,无显著性差异。
结论:SMO保存液对大鼠肝脏冷缺血损伤的保护效果与UW液类似,但其减轻冷缺血再灌注损伤的能力弱于UW液。
Chapter 1 The optimal molecular weight and concentration of PEG added to the newly developed SMO solution
Objective: To evaluate the optimal molecular weight and concentration of PEG added to the newly developed SMO solution.
Methods: Livers from SD rats were flushed and then preserved at 4°C for 24h in SMO solution with different molecular weight and concentration of PEG (no PEG, 1g/L PEG20, 10g/L PEG20, 30g/L PEG20, 1g/L PEG35, 10g/L PEG35 and 30g/L PEG35). At the end of the preservation, histological examination of hepatic tissues was performed. Then human venous blood was mixed with normal saline, UW solution, SMO solutions with different molecular weight and concentration of PEG. The dilution ratios of blood with preservation solution were 5:1 and 1:1. Sedimentation rate was measured by an automatic ESR analyzer. Human RBC aggregability and blood viscosity was investigated with an automatic hemorheological analyzer. Light microscopy was used to evaluate morphological characters of the RBC aggregates.
Results: After cold preservation for 24h, SMO solution with PEG35 (1g/L) showed the best preservation effect among the tested solution. PEG20 (1 and 10g/L) and PEG35 (1g/L) had little effect on RBC aggregation, while PEG20 (30g/L) and PEG35 (10 and 30g/L) significantly increased the RBC sedimentation and blood viscosity, and had a hyperaggregating effect on RBC.
Conclusion: Low concentration PEG35 (1g/L) would be the optimal choice added to the SMO solution.
Chapter 2 Establishment of the isolated perfused rat liver model and its modification
Objective: To set up the isolated perfused rat liver model, provide a reliable study platform for the development of organ preservation solution.
Methods: We modified the isolated perfused system described previously in references and establish CZ - 1 isolated perfused rat liver system. Then the effectiveness of the isolated perfused system was tested. Livers were harvested after the cannulation of the portal vein and bile duct. Then Livers were connected via the portal vein to a recirculating perfusion system for 120min. The reperfusion solution was Krebs-Henseleit solution at a constant temperature of 37°C and a flow rate of 20ml/min. After 120min reperfusion, bile production was evaluated. Routine HE staining examination of hepatic tissues was also performed.
Results: The CZ - 1 isolated perfused rat liver system was cost-effective and reliable to use. It was easy to run. The bile volume collected from the isolated repefused liver were 0.248±0.094 ul/min·g (liver) and was not significantly different from that reported by references. Hepatic tissues in reperfusion group were also morphologically normal.
Conclusion: The isolated perfused rat liver model closely mimics physiologic conditions and is the ideal model for investigation of organ preservation solution.
Chapter 3 Effects of SMO solution in rat liver preservation
Objective: To compare the protective effect of SMO solution with that of UW solution during cold preservation and normothermic reperfusion.
Methods: SD rats were divided into four groups according to different preservation solution: control group (without cold preservation), lactated Ringers group (negative control), UW group and SMO group. Preservation injury after cold storage for 24, 48 and 72h was assessed microscopically. Meanwhile the pH value and osmotic pressure of the preservation solutions was measured. The change of the liver weight was also assessed. After 24h cold storage of rat liver in different preservation solutions, the isolated perfused rat liver model was applied to reperfuse the liver for 120min normothermically with Krebs-Henseleit solution. Reperfusion injury was analyzed in the isolated perfused liver by enzyme (AST, ALT and LDH) release and bile production. Routine HE staining and electron microscopic examination of hepatic tissues were performed. NF-κB, ICAM-1 and Caspase-3 expressions were assessed by immunohistochemical analysis. Hepatocellular apoptosis was also assessed
Results: After preserving for 24h, histology showed no difference between the SMO and UW group. But after 48h cold preservation, UW solution provided rat livers with better protection against cold ischemia injury. During cold preservation, pH value of the SMO and UW solution decreased with preservation time, and the buffering capacity of both solutions was similar. The osmotic pressure of the both preservation solutions increased with preservation time, but they were higher in UW solution. Prolonged cold preaervation time resulted in an increase in liver weight with SMO-preserved livers and a decrease in weight with UW-preserved livers. After 24h cold storage, transaminase level in the both groups showed no difference. After reperfusion for 120min, ALT level was lower in SMO group, but higher AST and LDH levels were found in SMO group than in UW group. The amount of Bile product after reperfusion for 120min in UW group was more than that in SMO group. Morphological changes in SMO group were significiently worse than that in UW group. Histological results showed more necrotic regions in livers preserved in SMO solution. Electron microscopy revealed that ultrastructural impairments were similar in both groups. No significant differences were found in NF-κB and ICAM-1 expression between SMO group and UW group. The percentage of apoptotic cells and the expression patterns of apoptosis-related-genes were similar in livers preserved in SMO and UW solutions. The variables in both groups were better than those of livers preserved in Ringers solution.
Conclusion: The short-term cold preservation effect of SMO solution is similar with that of UW solution. However, SMO solution is inferior to UW solution in protecting the liver from cold ischemia-reperfusion injury.
引文
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[6] Hata K, Tolba RH, Wei L, et al. Impact of polysol, a newly developed preservation solution, on cold storage of steatotic rat livers. Liver Transpl, 2007, 13(1): 114-121.
[7] Billault C, Vaessen C, Van Glabeke E, et al. Use of the SCOT solution in kidney transplantation: preliminary report. Transplant Proc, 2006, 38(7): 2281-2282.
[8] Maathuis MH, Ottens PJ, van Goor H, et al. Static cold storage preservation of ischemically damaged kidneys. a comparison between IGL-1 and UW solution. Transpl Int, 2008, 21(5): 473-482.
[9] Neuzillet Y, Giraud S, Lagorce L, et al. Effects of the molecular weight of peg molecules (8, 20 and 35 KDA) on cell function and allograft survival prolongation in pancreatic islets transplantation. Transplant Proc, 2006, 38(7): 2354-2355.
[10] Mosbah IB, Saidane D, Peralta C, et al. Efficacy of polyethylene glycols in University of Wisconsin preservation solutions: a study of isolated perfused rat liver. Transplant Proc, 2005, 37(9): 3948-3950.
[1] Morariu AM, Vd Plaats A, V Oeveren W, et al. Hyperaggregating effect of hydroxyethyl starch components and University of Wisconsin solution on human red blood cells: a risk of impaired graft perfusion in organ procurement? Transplantation, 2003, 76(1): 37-43.
[2] Hata K, Tolba RH, Wei L, et al. Impact of polysol, a newly developed preservation solution, on cold storage of steatotic rat livers. Liver Transpl, 2007, 13(1): 114-121.
[3] Billault C, Vaessen C, Van Glabeke E, et al. Use of the SCOT solution in kidney transplantation: preliminary report. Transplant Proc, 2006, 38(7): 2281-2282.
[4] Maathuis MH, Ottens PJ, van Goor H, et al. Static cold storage preservation of ischemically damaged kidneys. a comparison between IGL-1 and UW solution. Transpl Int, 2008, 21(5): 473-482.
[5] van der Plaats A, 't Hart NA, Morariu AM, et al. Effect of University of Wisconsin organ-preservation solution on haemorheology. Transpl Int, 2004, 17(5): 227-33.
[6] Panzera P, Rotelli MT, Salerno AM, et al. Solutions for organ perfusion and storage: haemorheologic aspects. Transplant Proc, 2005, 37(6): 2456-2458.
[7] Gibelin H, Hauet T, Eugene M, et al. Beneficial effects of addition of polyethylene glycol to extracellular type solutions to minimize ischemia/reperfusion injuries in an isolated-perfused rat liver model. Transplant Proc, 2002, 34(3): 768.
[8] Dutheil D, Rioja-Pastor I, Tallineau C, et al. Protective effect of PEG 35,000 Da on renal cells: paradoxical activation of JNK signaling pathway during cold storage. Am J Transplant, 2006, 6(7): 1529-1540.
[9] Mosbah IB, Franco-Gou R, Abdennebi HB, et al. Effects of polyethylene glycol and hydroxyethyl starch in University of Wisconsin preservation solution on human red blood cell aggregation and viscosity. Transplant Proc, 2006, 38(5): 1229-1235.
[10] Chien S, Jan KM, et al. Red cell aggregation by macromolecules: roles of surface adsorption and electrostatic repulsion. J Supramol Struct, 1973, (4): 385-409.
[11]于戈群,阿丽亚,谷米,等.细胞的聚集性和血浆黏度及其蛋白组分在血液黏度调节中的作用.中国微循环,2004,8(5):298-300.
[12] Mosbah IB, Saidane D, Peralta C, et al. Efficacy of polyethylene glycols in University of Wisconsin preservation solutions: a study of isolated perfused rat liver. Transplant Proc, 2005, 37(9): 3948-3950.
[13] Neuzillet Y, Giraud S, Lagorce L, et al. Effects of the molecular weight of peg molecules (8, 20 and 35 KDA) on cell function and allograft survival prolongation in pancreatic islets transplantation. Transplant Proc, 2006, 38(7): 2354-2355.
[1] Ben Abdennebi H, Steghens JP, Margonari J, et al. High-Na+ low-K+ UW cold storage solution reduces reperfusion injuries of the rat liver graft. Transpl Int, 1998, 11(3): 223-230.
[2] Ben Mosbah I, Roselló-Catafau J, Franco-Gou R, et al. Preservation of steatotic livers in IGL-1 solution. Liver Transpl, 2006, 12(8): 1215-1223.
[3] Mosbah IB, Saidane D, Peralta C, et al. Efficacy of polyethylene glycols in University of Wisconsin preservation solutions: a study of isolated perfused rat liver. Transplant Proc, 2005, 37(9):3948-3950.
[4] Bessems M, 't Hart NA, Tolba R,et al. The isolated perfused rat liver: standardization of a time-honoured model. Lab Anim, 2006, 40(3): 236-246.
[5] Ontell SJ, Colella MS, Horowitz J, et al. Applications of the isolated perfused rat liver in transplantation research. J Invest Surg, 1988, 1(1): 25-27.
[6] Ahmed I, Attia MS, Ahmad N, et al. Use of isolated perfused rat liver model for testing liver preservation solutions. Transplant Proc, 2001, 33(7-8): 3709-3711.
[7] Defamie V, Laurens M, Patrono D, et al. Matrix metalloproteinase inhibition protects rat livers from prolonged cold ischemia-warm reperfusion injury. Hepatology, 2008, 47(1): 177-185.
[8] Boehnert MU, Armbruster FP, Hilbig H. Relaxin as a protective substance in preservation solutions for organ transplantation, as shown in an isolated perfused rat liver model. Transplant Proc, 2008, 40(4): 978-980.
[9] Cheng Y, Liu YF, Cheng DH, et al. Evaluation of CMU-1 preservation solutions using an isolated perfused rat liver model. World J Gastroenterol, 2005, 11(16): 2522-2525.
[10]徐东波,杜智,Potts DJ,等.大鼠离体肝再灌注模型(IPRL)的建立及其在肝移植器官保存液实验中的意义.生物医学工程与临床,2001,5(1):1-3.
[11]苏树炎,霍枫.大鼠自体肝移植中三种不同肝脏灌注方法的比较.肝胆胰外科杂志,2008,20(4):248-253.
[12] Gores GJ, Kost LJ, LaRusso JF. The isolated perfused rat liver conceptual and practical consideration. Hepatolgy, 1986, 6 (3): 5l1.
[13] 't Hart NA, van der Plaats A, Moers C , et al. Development of the isolated dual perfused rat liver model as an improved reperfusion model for transplantation research. Int J Artif Organs, 2006, 29(2): 219-227.
[1] Daemen JH, Heineman E, Kootstra G. Viability assessment of non-heart-beating donor kidneys during machine preservation. Transplant Proc, 1995, 27(5): 2906-2907.
[2] Southard JH, Belzer FO. Organ preservation. Annu Rev Med, 1995, 46: 235-247.
[3] McAnulty JF, Reid TW, Waller KR, et al. Successful six-day kidney preservation using trophic factor supplemented media and simple cold storage. Am J Transplant, 2002, 2(8): 712-718.
[4] Jamieson NV, Sundberg R, Lindell S, et al. Preservation of the canine liver for 24–48 hours using simple cold storage with UW solution. Transplantation, 1988, 46(4): 517-522.
[5] Grace P, Mathie R. Ischaemia-Reperfusion Injury. Boston: Blackwell Science; 1999.
[6] Bonventre JV, Cheung JY. Effects of metabolic acidosis on viability of cells exposed to anoxia. Am J Physiol, 1985, 249: C149-159.
[7] de Boer J, De Meester J, Smits JM, et al. Eurotransplant randomized multicenter kidney graft preservation study comparing HTK with UW and Euro-Collins. Transpl Int, 1999, 12(6): 447-453.
[8] Cavallari A, Cillo U, Nardo B, et al. A multicenter pilot prospective study comparing Celsior and University of Wisconsin preserving solutions for use in liver transplantation. Liver Transpl, 2003, 9(8): 814-821.
[9] Fridell JA, Agarwal A, Milgrom ML, et al. Comparison of histidinetryptophan-ketoglutarate solution and University of Wisconsin solution for organ preservation in clinical pancreas transplantation. Transplantation, 2004, 77(8): 1304-1306.
[10] Sumimoto R, Jamieson NV, Kamada N. Examination of the role of the impermeants lactobionate and raffinose in a modified UW solution. Transplantation, 1990, 50(4): 573-576.
[11] Jamieson NV, Lindell S, Sundberg R, et al. An analysis of the components in UW solution using the isolated perfused rabbit liver. Transplantation, 1988, 46(4): 512-516.
[12] Morariu AM, Vd Plaats A, V Oeveren W, et al. Hyperaggregating effect of hydroxyethyl starch components and University of Wisconsin solution on human red blood cells: a risk of impaired graft perfusion in organ procurement? Transplantation, 2003, 76(1): 37-43.
[13] Sankary HN, Foster P, Brown E, et al. A comparison of Collins and UW solutions for cold ischemic preservation of the rat liver. J Surg Res, 1991, 51(1):87-91.
[14] Sundberg R, Ar'rajab A, Ahrén B, et al. The functional effects of suppression of hypothermia-induced cell swelling in liver preservation by cold storage. Cryobiology, 1991, 28(2): 150-158.
[15] Momii S, Koga A. Time-related morphological changes in cold-stored rat livers. A comparison of Euro-Collins solution with UW solution. Transplantation, 1990, 50(5): 745-750.
[1] Ploeg RJ, D’Alessandro AM, Knechtle SJ, et al. Risk factors for primary dysfunction after liver transplantation-a multivariate analysis. Transplantation, 1993, 55(4): 807-813.
[2] Furukawa H, Todo S, Imventarza O, et al. Effect of cold ischemia time on the early outcome of human hepatic allografts preserved with UW solution. Transplantation, 1991, 51(5): 1000-1004.
[3] Lemasters JJ, Bunzendahl H, Thurman RG. Reperfusion injury to donor livers stored for transplantation. Liver Transpl Surg, 1995, 1(2): 124-138.
[4] Teoh NC, Farrell GC. Hepatic ischemia reperfusion injury: pathogenic mechanisms and basis for hepatoprotection. J Gastroenterol Hepatol, 2003, 18(8): 891-902.
[5] Arii S, Teramoto K, Kawamura T. Current progress in the understanding of and therapeutic strategies for ischemia and reperfusion injury of the liver. J Hepatobiliary Pancreat Surg, 2003, 10(3): 189-194.
[6] N.V. Jamieson, S. Lindell, R. Sundberg, et al. Belzer, An analysis of the components in UW solution using the isolated perfused rabbit liver, Transplantation 1988, 46 (4): 512–516.
[7] J. Kerr-Conte, K. Boudjema, J. Southard, et al. Mechanism of hypothermic cell death: glutathione prevents injury in hepatocytes during hypothermic (4℃) preservation. Transplant Proc 1991, 23 (5): 2405–2406.
[8] El-Wahsh M, Fuller B, Davidson B, et al. Hepatic cold hypoxia and oxidative stress: implications for ICAM-1 expression and modulation by glutathione during experimental isolated liver preservation. Cryobiology, 2003, 47(2):165-73.
[9] Ben Abdennebi H, Steghens JP, Margonari J, et al. High-Na+ low-K+ UW cold storage solution reduces reperfusion injuries of the rat liver graft. Transpl Int, 1998, 11(3): 223-230.
[10] Otto G, Wolff H, David H. Preservation damage in liver transplantation: electron-microscopic findings. Transplant Proc, 1984, 16(5): 1247-1248.
[11] Clavien PA, Harvey PR, Strasberg SM. Preservation and reperfusion injuries in liver allografts. An overview and synthesis of current studies. Transplantation,1992, 53: 957-978.
[12] Kohli V, Selzner M, Madden JF,et al. Endothelial cell and hepatocyte deaths occur by apoptosis after ischemiareperfusion injury in the rat liver. Transplantation 1999, 67:1099-1105.
[13] Kam PC, Ferch NI. Apoptosis: mechanisms and clinical implications. Anaesthesia, 2000, 55(11): 1081-1093.
[14] Duan WR, Garner DS, Williams SD, et al. Comparison of immunohistochemistry for activated caspase-3 and cleaved cytokeratin 18 with the TUNEL method for quantification of apoptosis in histological sections of PC-3 subcutaneous xenografts. J Pathol, 2003, 199 (2): 221-228.
[15] Gujral JS, Bucci TJ, Farhood A, et al. Mechanism of cell death during hepatic ischemia-reperfusion in rats: apoptosis or necrosis? Hepatology 2001, 33(2): 397–405.
[16] Jaeschke H, Lemasters JJ. Apoptosis versus oncotic necrosis in hepatic ischemia/reperfusion injury. Gastroenterology, 2003, 125(4): 1246-1257.
[17] Ali S, Mann DA. Signal transduction via the NF-kappaB pathway: a targeted treatment modality for infection, inflammation and repair. Cell Biochem Funct, 2004, 22(2): 67-79.
[18] Gu XP, Jiang Y, Xu FT, et al. Effect of cold-ischemia time on nuclear factor-kappaB activation and inflammatory response in graft after orthotopic liver transplantation in rats. World J Gastroenterol, 2004, 10(7): 1000-1004.
[19] M. Morita, Y. Watanabe, T. Akaike. Inflammatory cytokines up-regulate intercellular adhesion molecule-1 expression on primary cultured mouse hepatocytes and T lymphocyte adhesion. Hepatology, 1994, 19 (2): 426–431.
[20] M.L. Dustin, T.A. Springer. Lymphocyte function-associated antigen-1 (LFA-1) interaction with intercellular adhesion molecule-1 (ICAM-1) is one of at least three mechanisms for lymphocyte adhesion to cultured endothelial cells. J Cell Biol, 1988, 107 (1): 321–331.
[1] Daniel MR, Wakerley CL. Factors influencing the survival of cell monolayers during storage at 4 degrees. Br J Exp Pathol, 1976, 57(2): 137-147.
[2] Hauet T, Baumert H, Amor IB, et al. Protection of autotransplanted pig kidneys from ischemia-reperfusion injury by polyethylene glycol. Transplantation, 2000, 70(11): 1569-1575.
[3] Gibelin H, Hauet T, Eugene M, et al. Beneficial effects of addition of polyethylene glycol to extracellular type solutions to minimize ischemia/reperfusion injuries in an isolated-perfused rat liver model. Transplant Proc, 2002, 34(3): 768.
[4] Jayle C, Corbi P, Eugene M, et al. Beneficial effect of polyethylene glycol in lung preservation: early evaluation by proton nuclear magnetic resonance spectroscopy. Ann Thorac Surg, 2003, 76(3): 896-902.
[5] Dutheil D, Rioja-Pastor I, Tallineau C, et al. Protective effect of PEG 35,000 Da on renal cells: paradoxical activation of JNK signaling pathway during cold storage. Am J Transplant, 2006, 6(7): 1529-1540.
[6] Mosbah IB, Saidane D, Peralta C, et al. Efficacy of polyethylene glycols in University of Wisconsin preservation solutions: a study of isolated perfused rat liver. Transplant Proc, 2005, 37(9): 3948-3950.
[7] Neuzillet Y, Giraud S, Lagorce L, et al. Effects of the molecular weight of peg molecules (8, 20 and 35 KDA) on cell function and allograft survival prolongation in pancreatic islets transplantation. Transplant Proc, 2006, 38(7): 2354-2355.
[8] Bessems M, Doorschodt BM, van Vliet AK, et al. Machine perfusion preservation of the non-heart-beating donor rat livers using polysol, a new preservation solution [J]. Transplant Proc, 2005, 37(1): 326-328.
[9] Hata K, Tolba RH, Wei L, et al. Impact of polysol, a newly developed preservation solution, on cold storage of steatotic rat livers. Liver Transpl, 2007, 13(1): 114-121.
[10] Wei L, Hata K, Doorschodt BM, et al. Experimental small bowel preservation using Polysol: a new alternative to University of Wisconsin solution, Celsior and histidine-tryptophan-ketoglutarate solution? World J Gastroenterol, 2007, 13(27): 3684-3691.
[11] Billault C, Vaessen C, Van Glabeke E, et al. Use of the SCOT solution in kidney transplantation: preliminary report. Transplant Proc, 2006, 38(7): 2281-2282.
[12] Maathuis MH, Ottens PJ, van Goor H, et al. Static cold storage preservation of ischemically damaged kidneys. a comparison between IGL-1 and UW solution. Transpl Int, 2008, 21(5): 473-482.
[13] Stefanovich P, Ezzell RM, Sheehan SJ, et al. Effects of hypothermia on the function, membrane integrity, and cytoskeletal structure of hepatocytes. Cryobiology, 1995, 32(4): 389-403.
[14] Hauet T, Mothes D, Goujon JM, et al. Protective effect of polyethylene glycol against prolonged cold ischemia and reperfusion injury: study in the isolated perfused rat kidney. J Pharmacol Exp Ther, 2001, 297(3): 946-952.
[15] Mosbah IB, Franco-Gou R, Abdennebi HB, et al. Effects of polyethylene glycol and hydroxyethyl starch in University of Wisconsin preservation solution on human red blood cell aggregation and viscosity. Transplant Proc, 2006, 38(5): 1229-1235.
[16] Murad KL, Gosselin EJ, Eaton JW, et al. Stealth cells: prevention of major histocompatibility complex class II-mediated T-cell activation by cell surface modification. Blood, 1999, 94(6): 2135-2141.
[17] Murad KL, Mahany KL, Brugnara C, et al. Structural and functional consequences of antigenic modulation of red blood cells with methoxypoly(ethylene glycol). Blood, 1999, 93(6): 2121-2127.
[18] Stuhlmeier KM, Lin Y. Camouflaging endothelial cells: does it prolong graft survival? Biochim Biophys Acta, 1999, 1428(2): 177-190.
[2] Belzer FO, Southard JH. Principles of solid-organ preservation by cold storage. Transplantation, 1988, 45(4): 673-676.
[3] Collins GM, Bravo-Shugarman M, Terasaki PI. Kidney preservation for transportation. Initial perfusion and 30 hours’ice storage. Lancet, 1969, 2(7632): 1219-1222.
[4] Annual Report Eurotransplant International Foundation. Leiden. The Netherlands: Eurotransplant International Foundation, 1976.
[5] Hoffman B, Sollinger H, Kalayoglu M, et al. Use of UW solution for kidney transplantation. Transplantation, 1988, 46(2): 338-339.
[6] Bretschneider HJ. Myocardial protection. Thorac Cardiovasc Surg, 1980, 28(5): 295-302.
[7] Roels L, Coosemans W, Donck J, et al. Inferior outcome of cadaveric kidneys preserved for more than 24 hr in histidine-tryptophanketoglutarate solution. Leuven Collaborative Group for Transplantation. Transplantation, 1998, 66(12): 1660-1664.
[8] Menasche P, Termignon JL, Pradier F, et al. Experimental evaluation of Celsior, a new heart preservation solution. Eur J Cardiothorac Surg, 1994, 8(4): 207-213.
[9] Janssen H, Janssen PH, Broelsch CE. Celsior solution compared with University of Wisconsin solution (UW) and histidine-tryptophanketoglutarate solution (HTK) in the protection of human hepatocytes against ischemia-reperfusion injury. Transpl Int 2003; 16(7): 515-522.
[10] Bessems M, Doorschodt BM, van Vliet AK, et al. Machine perfusion preservation of the non-heart-beating donor rat livers using polysol, a new preservation solution. Transplant Proc, 2005, 37(1): 326-328.
[11] Hata K, Tolba RH, Wei L, et al. Impact of polysol, a newly developed preservation solution, on cold storage of steatotic rat livers. Liver Transpl, 2007, 13(1): 114-121.
[12] Wei L, Hata K, Doorschodt BM, et al. Experimental small bowel preservation using Polysol: a new alternative to University of Wisconsin solution, Celsior and histidine-tryptophan-ketoglutarate solution? World J Gastroenterol, 2007, 13(27): 3684-3691.
[13] Billault C, Vaessen C, Van Glabeke E, et al. Use of the SCOT solution in kidneytransplantation: preliminary report. Transplant Proc, 2006, 38(7): 2281-2282.
[14] Ben Abdennebi H, Steghens JP, Hadj-Aissa A, et al. A preservation solution with polyethylene glycol and calcium: A possible multiorgan liquid. Transpl Int, 2002, 15(7): 348-354.
[15] Maathuis MH, Ottens PJ, van Goor H, et al. Static cold storage preservation of ischemically damaged kidneys. a comparison between IGL-1 and UW solution. Transpl Int, 2008, 21(5): 473-482.
[1] Daniel MR, Wakerley CL. Factors influencing the survival of cell monolayers during storage at 4 degrees. Br J Exp Pathol, 1976, 57(2): 137-147.
[2] Hauet T, Baumert H, Amor IB, et al. Protection of autotransplanted pig kidneys from ischemia-reperfusion injury by polyethylene glycol. Transplantation, 2000, 70(11): 1569-1575.
[3] Gibelin H, Hauet T, Eugene M, et al. Beneficial effects of addition of polyethylene glycol to extracellular type solutions to minimize ischemia/reperfusion injuries in an isolated-perfused rat liver model. Transplant Proc, 2002, 34(3): 768.
[4] Jayle C, Corbi P, Eugene M, et al. Beneficial effect of polyethylene glycol in lung preservation: early evaluation by proton nuclear magnetic resonance spectroscopy. Ann Thorac Surg, 2003, 76(3): 896-902.
[5] Dutheil D, Rioja-Pastor I, Tallineau C, et al. Protective effect of PEG 35,000 Da on renal cells: paradoxical activation of JNK signaling pathway during cold storage. Am J Transplant, 2006, 6(7): 1529-1540.
[6] Hata K, Tolba RH, Wei L, et al. Impact of polysol, a newly developed preservation solution, on cold storage of steatotic rat livers. Liver Transpl, 2007, 13(1): 114-121.
[7] Billault C, Vaessen C, Van Glabeke E, et al. Use of the SCOT solution in kidney transplantation: preliminary report. Transplant Proc, 2006, 38(7): 2281-2282.
[8] Maathuis MH, Ottens PJ, van Goor H, et al. Static cold storage preservation of ischemically damaged kidneys. a comparison between IGL-1 and UW solution. Transpl Int, 2008, 21(5): 473-482.
[9] Neuzillet Y, Giraud S, Lagorce L, et al. Effects of the molecular weight of peg molecules (8, 20 and 35 KDA) on cell function and allograft survival prolongation in pancreatic islets transplantation. Transplant Proc, 2006, 38(7): 2354-2355.
[10] Mosbah IB, Saidane D, Peralta C, et al. Efficacy of polyethylene glycols in University of Wisconsin preservation solutions: a study of isolated perfused rat liver. Transplant Proc, 2005, 37(9): 3948-3950.
[1] Morariu AM, Vd Plaats A, V Oeveren W, et al. Hyperaggregating effect of hydroxyethyl starch components and University of Wisconsin solution on human red blood cells: a risk of impaired graft perfusion in organ procurement? Transplantation, 2003, 76(1): 37-43.
[2] Hata K, Tolba RH, Wei L, et al. Impact of polysol, a newly developed preservation solution, on cold storage of steatotic rat livers. Liver Transpl, 2007, 13(1): 114-121.
[3] Billault C, Vaessen C, Van Glabeke E, et al. Use of the SCOT solution in kidney transplantation: preliminary report. Transplant Proc, 2006, 38(7): 2281-2282.
[4] Maathuis MH, Ottens PJ, van Goor H, et al. Static cold storage preservation of ischemically damaged kidneys. a comparison between IGL-1 and UW solution. Transpl Int, 2008, 21(5): 473-482.
[5] van der Plaats A, 't Hart NA, Morariu AM, et al. Effect of University of Wisconsin organ-preservation solution on haemorheology. Transpl Int, 2004, 17(5): 227-33.
[6] Panzera P, Rotelli MT, Salerno AM, et al. Solutions for organ perfusion and storage: haemorheologic aspects. Transplant Proc, 2005, 37(6): 2456-2458.
[7] Gibelin H, Hauet T, Eugene M, et al. Beneficial effects of addition of polyethylene glycol to extracellular type solutions to minimize ischemia/reperfusion injuries in an isolated-perfused rat liver model. Transplant Proc, 2002, 34(3): 768.
[8] Dutheil D, Rioja-Pastor I, Tallineau C, et al. Protective effect of PEG 35,000 Da on renal cells: paradoxical activation of JNK signaling pathway during cold storage. Am J Transplant, 2006, 6(7): 1529-1540.
[9] Mosbah IB, Franco-Gou R, Abdennebi HB, et al. Effects of polyethylene glycol and hydroxyethyl starch in University of Wisconsin preservation solution on human red blood cell aggregation and viscosity. Transplant Proc, 2006, 38(5): 1229-1235.
[10] Chien S, Jan KM, et al. Red cell aggregation by macromolecules: roles of surface adsorption and electrostatic repulsion. J Supramol Struct, 1973, (4): 385-409.
[11]于戈群,阿丽亚,谷米,等.细胞的聚集性和血浆黏度及其蛋白组分在血液黏度调节中的作用.中国微循环,2004,8(5):298-300.
[12] Mosbah IB, Saidane D, Peralta C, et al. Efficacy of polyethylene glycols in University of Wisconsin preservation solutions: a study of isolated perfused rat liver. Transplant Proc, 2005, 37(9): 3948-3950.
[13] Neuzillet Y, Giraud S, Lagorce L, et al. Effects of the molecular weight of peg molecules (8, 20 and 35 KDA) on cell function and allograft survival prolongation in pancreatic islets transplantation. Transplant Proc, 2006, 38(7): 2354-2355.
[1] Ben Abdennebi H, Steghens JP, Margonari J, et al. High-Na+ low-K+ UW cold storage solution reduces reperfusion injuries of the rat liver graft. Transpl Int, 1998, 11(3): 223-230.
[2] Ben Mosbah I, Roselló-Catafau J, Franco-Gou R, et al. Preservation of steatotic livers in IGL-1 solution. Liver Transpl, 2006, 12(8): 1215-1223.
[3] Mosbah IB, Saidane D, Peralta C, et al. Efficacy of polyethylene glycols in University of Wisconsin preservation solutions: a study of isolated perfused rat liver. Transplant Proc, 2005, 37(9):3948-3950.
[4] Bessems M, 't Hart NA, Tolba R,et al. The isolated perfused rat liver: standardization of a time-honoured model. Lab Anim, 2006, 40(3): 236-246.
[5] Ontell SJ, Colella MS, Horowitz J, et al. Applications of the isolated perfused rat liver in transplantation research. J Invest Surg, 1988, 1(1): 25-27.
[6] Ahmed I, Attia MS, Ahmad N, et al. Use of isolated perfused rat liver model for testing liver preservation solutions. Transplant Proc, 2001, 33(7-8): 3709-3711.
[7] Defamie V, Laurens M, Patrono D, et al. Matrix metalloproteinase inhibition protects rat livers from prolonged cold ischemia-warm reperfusion injury. Hepatology, 2008, 47(1): 177-185.
[8] Boehnert MU, Armbruster FP, Hilbig H. Relaxin as a protective substance in preservation solutions for organ transplantation, as shown in an isolated perfused rat liver model. Transplant Proc, 2008, 40(4): 978-980.
[9] Cheng Y, Liu YF, Cheng DH, et al. Evaluation of CMU-1 preservation solutions using an isolated perfused rat liver model. World J Gastroenterol, 2005, 11(16): 2522-2525.
[10]徐东波,杜智,Potts DJ,等.大鼠离体肝再灌注模型(IPRL)的建立及其在肝移植器官保存液实验中的意义.生物医学工程与临床,2001,5(1):1-3.
[11]苏树炎,霍枫.大鼠自体肝移植中三种不同肝脏灌注方法的比较.肝胆胰外科杂志,2008,20(4):248-253.
[12] Gores GJ, Kost LJ, LaRusso JF. The isolated perfused rat liver conceptual and practical consideration. Hepatolgy, 1986, 6 (3): 5l1.
[13] 't Hart NA, van der Plaats A, Moers C , et al. Development of the isolated dual perfused rat liver model as an improved reperfusion model for transplantation research. Int J Artif Organs, 2006, 29(2): 219-227.
[1] Daemen JH, Heineman E, Kootstra G. Viability assessment of non-heart-beating donor kidneys during machine preservation. Transplant Proc, 1995, 27(5): 2906-2907.
[2] Southard JH, Belzer FO. Organ preservation. Annu Rev Med, 1995, 46: 235-247.
[3] McAnulty JF, Reid TW, Waller KR, et al. Successful six-day kidney preservation using trophic factor supplemented media and simple cold storage. Am J Transplant, 2002, 2(8): 712-718.
[4] Jamieson NV, Sundberg R, Lindell S, et al. Preservation of the canine liver for 24–48 hours using simple cold storage with UW solution. Transplantation, 1988, 46(4): 517-522.
[5] Grace P, Mathie R. Ischaemia-Reperfusion Injury. Boston: Blackwell Science; 1999.
[6] Bonventre JV, Cheung JY. Effects of metabolic acidosis on viability of cells exposed to anoxia. Am J Physiol, 1985, 249: C149-159.
[7] de Boer J, De Meester J, Smits JM, et al. Eurotransplant randomized multicenter kidney graft preservation study comparing HTK with UW and Euro-Collins. Transpl Int, 1999, 12(6): 447-453.
[8] Cavallari A, Cillo U, Nardo B, et al. A multicenter pilot prospective study comparing Celsior and University of Wisconsin preserving solutions for use in liver transplantation. Liver Transpl, 2003, 9(8): 814-821.
[9] Fridell JA, Agarwal A, Milgrom ML, et al. Comparison of histidinetryptophan-ketoglutarate solution and University of Wisconsin solution for organ preservation in clinical pancreas transplantation. Transplantation, 2004, 77(8): 1304-1306.
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