阿霉素预处理对大鼠供肝热缺血再灌注损伤的保护作用

目的探讨阿霉素预处理对无心跳大鼠供肝热缺血再灌注损伤的保护作用。方法根据阿霉素预处理与否及供肝获取前经历的供体心脏停搏时间30min或45min，将实验动物分为4组，即非预处理的30min（N-30）组和45min（N-45）组，阿霉素预处理的30min（tN-30）组和45min（tN-45）组行原位肝移植，观察存活状况，取材行光学显微镜及电子显微镜检查，移植术后1、3、7d采集血样检测肝功能。结果N-30组和N-45组的1周存活率分别为50．0％和16．7％，而tN-30组和tN-45组的1周存活率分别为83．3％和66．7％，预处理组移植肝脏的病理及肝功能明显好于非预处理组。结论阿霉素预处理能够减轻供肝的热缺血再灌注损伤。改善肝功能，减轻病理损害，提高大鼠肝移植的存活率。     

外源性三磷酸腺苷-氯化镁预处理对大鼠供肝热缺血损伤的保护作用

目的 观察三磷酸腺苷-氯化镁(ATP-MgCl2)预处理对无心跳大鼠供肝热缺血损伤的保护作用.方法 根据ATP-MgCl2预处理与否及供肝获取前经历的供体心脏停搏时间(即热缺血时间)30min或45min,将实验动物分为4组,即非预处理的30min(N-30min)组和45min(N-45min)组,以及ATP-MgCl2预处理的30min(tN-30min)组和45min(tN-45min)组行原位肝移植,观察存活状况,取材行光学显微镜及电子显微镜检查,移植术后1、3、7d 采集血样检测肝功能.结果 N-30min组和N-45min组的1周存活率分别为50.0%和16.7%,而tN-30min组和tN-45min组的1周存活率分别为83.3%和66.7%,预处理组移植肝脏的病理及肝功能明显好于非预处理组.结论 ATP-MgCl2预处理能够减轻供肝的热缺血损伤,改善肝功能,减轻病理损害,提高大鼠肝移植的存活率.     

大鼠心跳停搏供肝在原位肝移植术中损伤的预防

目的：探讨预防和减轻大鼠心跳停搏供肝在原位肝移植术中的损伤，以提高手术成功率。方法：雄性SD大鼠随机分为心跳停搏热缺血30min(N-30)和45min(N-45)两组;，每组分别行原位肝移植术30只次。同时，根据是否对供体手术方法进行改进又分为常规组和改良组。结果：(1)常规组和改良组的冷缺血时间分别为(70.04±1.48)和(70.36±1.42)min(P>0.05)，无肝期均为(16.40±0.73)min，肝下下腔静脉阻断时间均为(22.75±1.16)min，受体手术时间均为(90.58±3.76)min。(2)N-30和N-45常规组分别有5和9只受体术后死于原发性移植肝无功能，而改良组仅为1和2只(40%∶12%,P<0.05);(3)N-30和N-45组因术中分别出现供肝损伤致再灌注后供肝大量渗血、无肝期过长、切除受体肝脏时麻醉过深，而各有5和7，2和1，2和2只受体术后死亡。(4)N-30和N-45组术后1周存活率分别为50%和30%(P<0.05)。结论：预防心跳停搏供肝游离时损伤、供肝再灌注后渗血、无肝期过长和切除受体肝脏时麻醉过深是大鼠心跳停搏供肝原位肝移植手术成功的关键。     

Lazaroid U-74389G预处理对大鼠心脏停搏供体肝移植缺血再灌注损伤的保护作用

目的探讨在心脏停搏供体肝移植中 ,LazaroidU 74 389G预处理对大鼠供肝热缺血损伤的保护作用。方法实验以U 74 389G预处理与否、以及供肝获取前经历的心脏停搏时间 4 5或6 0min分为 4组 ,分别为N 4 5组、N 6 0组、tN 4 5组和tN 6 0组 ,而后行大鼠原位肝移植 ,比较各组术后存活率、肝功能、MDA和肝脏病理学改变。结果N 4 5组、N 6 0组、tN 4 5组和tN 6 0组肝移植术后1周的存活率分别为 2 5 % (2 / 8)、0 (0 / 8)、5 8% (7/ 12 )和 33% (4/ 12 )。U 74 389G的预处理可以显著提高心脏停搏供体肝移植的存活率 ,并能够改善术后肝功能和减轻肝脏病理学改变 ,降低MDA的表达。结论在心脏停搏供体肝移植中 ,U 74 389G预处理可以减轻热缺血再灌注对供肝造成的损伤 ,提高肝移植存活率     

肝移植热缺血损伤与细胞因子表达的研究

目的　探讨心脏停搏供体肝移植中 ,供肝热缺血损伤与肝移植术后细胞因子的表达。方法　将实验鼠以供肝获取前经历的心脏停搏时间 0、30和 6 0min分为 3组 ,每组 2 0只 ,分别为HB组、NHBD 30组和NHBD 6 0组 ,而后行大鼠原位肝移植。比较术后各组肝移植大鼠血清中TNF α、IL 6和CINC的变化。结果　随着供肝遭受的热缺血时间延长 ,肝移植大鼠术后血清中TNF α、IL 6和CINC均呈上升趋势 ,并分别于术后 3、6和 6h达到峰值 ,各组间差异有显著意义 (P <0 0 5 )。结论　在心脏停搏供体肝移植中 ,供肝的热缺血损伤与TNF α、IL 6和CINC的表达上调有关     

供肝热缺血与肝移植术后肺损伤关系的实验研究

目的：探讨心脏停搏供体肝移植中，供肝热缺血与肝移植术后肺损伤的关系。方法：实验以供肝获取前经历的心脏停搏时间0、30和60min分为3组，分别为有心跳组（HB组）、心脏停搏30min组（NHBD－30组）和心跳停搏60min组（NHBD－60组），而后行大鼠原位肝移植。比较术后血清中肿瘤坏死因子（TNF）－α细胞间粘附分子－1（ICAM－1）的变化，并观察肺组织病理和肺组织中ICAM－1免疫组织化学表达。结果：随着供肝热缺血时间的延长，肺组织损害也逐渐回重，术后血清中TNF－α和ICAM－1的值呈上升趋势，各组间差异有显著性。术后6h NHBD-60组出现肺组织中ICAM－1的强表达。结论：在心脏停搏供体肝移植中，TNF－α和ICAM－1的表达上调与术后肺组织损伤有关。     

增加供肝的肝糖原贮备减轻心脏停搏大鼠供肝的热缺血再灌注损伤

目的探讨肝移植术前增加供肝的肝糖原贮备能否减轻心脏停搏大鼠供肝的热缺血再灌注损伤。方法雄性SD大鼠作为肝移植的供、受者被随机分为A、B、C三组。A组供者正常饮水；B组供者术前连续4d饮糖水；C组供者在B组的基础上于供肝获取前3～4h注射500g／L的葡萄糖。A、B、C三组再按供者经历的心脏停搏时间（心脏停搏60min、90min、120min和150min）各分为4个小组，行原位肝移植术并检测移植肝组织中肝糖原和ATP的含量；同时观察受者术后1周存活率以及血清丙二醛（MDA）和超氧化物歧化酶（SOD）的水平。结果B、C两组供肝的肝糖原贮备和ATP含量均明显高于A组（P〈0．01）；B-60min、B-90min、B-120min和C-90min、C-120min、C-150min组受者的1周存活率分别为80％、50％、1（）％和70％、60％、20％，部分受者可长期存活；B、C两组受者MDA水平明显低于A组，SOD水平明显高于A组（P〈0.05）。结论术前增加大鼠供肝的肝糖原贮备能明显减轻供肝的热缺血再灌注损伤，减少了术后原发性移植肝无功能的发生，提高了心脏停搏供肝移植术后的存活率。     

肝移植时供肝耐受无心跳热缺血的安全时限实验研究

目的 探讨肝移植时供肝耐受无心跳热缺血损伤的安全时限。方法 建立广西巴马小型猪原位肝移植动物模型并按移植前供肝经历心脏停搏时间0、30、45、60min分为4组，观测肝移植后各组一周存活率、肝功能、肝脏病理和肝脏能量代谢以及术中肝脏复流后微循环改变。结果上述各组术后一周存活率分别为：100％(5／5)、100％(5／5)、60％(3／5)、20％(1／5)。肝脏能量代谢等指标的改变在供肝无心跳热缺血时间30min前是可逆的，随心跳热缺血时间的延长远渐向不可逆演变。结论 在本实验条件下，巴马小型猪心脏停搏供体肝移植时供肝耐受无心跳热缺血损伤的安全时限约为30min。     

不同热缺血时间对大鼠移植肝功能状态及存活期的影响

目的：探讨不同热缺血时间条件下，大鼠原位肝移植术后肝脏功能和组织形态变化规律，并对动物远期存活情况进行观察，预测无心跳供体（NHBD）中供肝耐受热缺血的安全时限。方法：实验动物按供肝热缺血时间分别为0、10、15、20、30、45、60min，随机分为7组。然后按各组条件分别作原位肝移植，观察肝移植后24、48h、3和5d各组肝脏功能和组织形态的恢复性变化，并统计生存时间。结果：供肝经受30min以内热缺血损伤，各组移植术后生存天数差异无显著性，热缺血30min组，肝移植术后1周的存活率达到83.3％（10/12），1个月和3个月存活率分别为58.3％（7/12）和50.0％（6/12）。热缺血45min组，术后1周的存活率为66.7％（8/12），但1个月和3个月存活率分别为33.3％（4/12）和8.3％（1/12）。至于热缺血60min组，1周的存活率仅为8.3％（1/12）。结论：大鼠的供肝可以安全地耐受30min以内的热缺血。45min的热缺血损伤，虽对术后1周存活率无明显影响，但大鼠肝移植术后的远期存活时间明显缩短。60min以后，移植肝组织的结构和功能均难以恢复，大鼠术后生存天数显著降低。     

供体热缺血时间对移植肝的影响

目的 探讨大鼠动脉化原位肝移植中供体热缺血时间对移植肝的影响.方法 实验分为4组:对照组(C)和移植组,移植组根据供肝获取前经历供体心脏停搏时间的不同分为三组:热缺血0 min(W0)、热缺血15 min(W15)和热缺血30 min(W30),其后建立近交系大鼠动脉化原位肝移植模型,每组均为30只大鼠,分别于术后3、7、14和30 d处死,每个时间点各取6只大鼠,分别测定移植肝组织学、肝功能的变化.此外,移植组各组随机选取6只大鼠观察长期生存率(＞100 d).结果 随着供肝热缺血时间的延长,移植肝损伤加重,恢复过程延长.移植组和对照组术后3、7、14和30 d血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)变化无显著性差异.血清碱性磷酸酶(ALP)随着供肝热缺血时间的延长逐渐增高,14 d达到高峰后逐渐下降.术后3、7、14和30 d ALP与供肝热缺血时间具有显著相关性.术后热缺血0、15、30 min长期生存率组分别为100.0%(6/6)、83.3%(5/6)、66.7%(4/6),3组间比较差异无统计学意义(P=0.285).结论 肝移植过程中供肝热缺血主要损伤肝细胞,并随着供肝热缺血时间的延长移植肝细胞损伤加重,肝细胞功能恢复早于其形态学恢复.肝移植术后早期存在胆汁淤积,供肝热缺血时间的延长明显加重胆汁淤积的程度,胆汁淤积的恢复明显晚于肝细胞损伤指标的恢复.在热缺血30 min内来自于心脏停搏的供肝肝移植术后是安全的.     

Preliminary study of rat liver transplantation from non‐heart‐beating donors

Objective : The aim of this study was to investigate the tolerance of warm ischaemia of liver grafts from non‐heart‐beating donors (NHBD). Livers harvested from NHBD would greatly expand the donor pool for transplantation. However, sensitivity of the donor liver to warm ischaemia is a major obstacle to the successful use of livers from NHBD and the limit of non‐heart‐beating time in the donor remains unclear. Materials and Methods : Rat orthotopic liver transplantation was performed in 5 groups (groups 1–5) with non‐heart‐beating time in the donor ranging from zero to 15, 30, 45 and 60 min. The cardiac‐arrest time was counted from clamping the base of the donor’s heart to the beginning of cold flushing the liver. Graft pathological changes, hepatic function and the recipients’ survival rates in each group were compared. Results : With a stepwise increase of the non‐heart‐beating time from zero to 15, 30, 45 and 60 min in donors, the recipient 1‐week survival rates from groups 1–5 were 100%, 75%, 62.5%, 25% and 0%, respectively. The stepwise increase of non‐heart‐beating time significantly reduced recipient survival rates after 30 min. The recipients in groups 1–4 could survive more than 60 days. Conclusions : The liver is less sensitive to warm ischaemia than was formerly believed. Rat liver can be used for transplantation after cardiac arrest for up to 45 min with a chance of survival.      

Novel short-term hypothermic oxygenated perfusion (HOPE) system prevents injury in rat liver graft from non-heart beating donor

OBJECTIVE: To assess a machine perfusion system in rescuing liver grafts from non-heart-beating donors (NHBD). SUMMARY BACKGROUND DATA: The introduction of extracorporeal liver perfusion systems in the clinical routine depends on feasibility. Conceivably, perfusion could be performed during recipient preparation. We investigated whether a novel rat liver machine perfusion applied after in situ ischemia and cold storage can rescue NHBD liver grafts. METHODS: We induced cardiac arrest in male Brown Norway rats by phrenotomy and ligation of the subcardial aorta. We studied 2 experimental groups: 45 minutes of warm in situ ischemia + 5 hours cold storage versus 45 minutes of warm in situ ischemia + 5 hours cold storage followed by 1 hour hypothermic oxygenated extracorporeal perfusion (HOPE). In both groups, livers were reperfused in a closed sanguineous isolated liver perfusion device for 3 hours at 37 degrees C. To test the benefit of HOPE on survival, we performed orthotopic liver transplantation in both experimental groups. RESULTS: After cold storage and reperfusion, NHBD livers showed necrosis of hepatocytes, increased release of AST, and decreased bile flow. HOPE improved NHBD livers significantly with a reduction of necrosis, less AST release, and increased bile flow. ATP was severely depleted in cold-stored NHBD livers but restored in livers treated by HOPE. After orthotopic liver transplantation, grafts treated by HOPE demonstrated a significant extension on animal survival. CONCLUSIONS: We demonstrate a beneficial effect of HOPE by preventing reperfusion injury in a clinically relevant NHBD model.     

不同热缺血时间的无心跳供肝在肝移植中应用的疗效评价

目的:探讨不同热缺血时间的无心跳供肝在临床肝移植中应用的疗效。方法:回顾性分析142例冷保存时间在12h之内的不同热缺血时间的无心跳供肝肝移植患者的临床资料, 根据热缺血时间分为3组：Ⅰ组为5 min之内, 43例; Ⅱ组为5～10 min, 77例; Ⅲ组为10～15 min, 22例。比较3组间肝移植术后谷丙转氨酶（ALT）峰值、原发性移植肝无功能、急性排斥反应、胆道并发症、血管并发症、感染以及移植肝存活和受体存活的差异。结果:3组患者术后均未发生原发性移植肝无功能。Ⅱ组术后ALT峰值及感染发生率显著高于Ⅰ组; 而Ⅲ组术后ALT峰值、胆道并发症发生率、血管并发症发生率及感染发生率均显著升高。随访 8~32个月。Ⅲ组移植肝存活率及受体存活率均降低, 与Ⅰ组和Ⅱ组比较, 差异均有统计学意义。结论:冷保存时间在12 h之内的无心跳供肝能耐受热缺血的安全时限为10 min; 热缺血时间大于10 min的供肝, 移植术后相关并发症发生率显著增加, 移植肝和受体存活率显著降低。     

Liver transplantation using non-heart-beating donors: Belgian experience

Mortality on liver transplantation (OLT) waiting lists has increased dramatically. Until recently, non-heart-beating donors (NHBD) were not considered suitable for OLT, because of a higher risk of primary graft nonfunction (PNF) and biliary strictures. However, recent experimental/clinical evidence has indicated that NHBD-OLT is feasible when the period of warm ischemia is short. PURPOSE: To characterize the results of NHBD-OLT in Belgium, a survey was sent to all Belgian OLT centers. RESULTS: Between January 2003 and November 2005, 16 livers originating from NHBD were procured and transplanted. The mean donor age was 48.8 years, including 9 males and 7 females with mean time of stop-therapy to cardiac arrest being 18 minutes and from cardiac arrest to liver cold perfusion, 10.5 minutes. Mean recipient age was 52.2 years including 12 males and 4 females. Mean cold ischemia time was 7 hours 15 minutes. No PNF requiring re-OLT was observed. Mean post-OLT peak transaminase was 2209 IU/L, which was higher among imported versus locally procured grafts. Biliary complications occurred in 6 patients requiring re-OLT (n = 2), endoscopic treatment (n = 2), surgical treatment (n = 1), or left untreated (n = 1). These tended to be more frequent after prolonged warm ischemia. Graft and patient survivals were 62.5% and 81.3%, respectively, with a follow-up of 3 to 36 months. CONCLUSION: This survey showed acceptable graft/patient survivals after NHBD-LT. The NHBD-liver grafts suffered a high rate of ischemic injury and biliary complications and therefore should be used carefully, namely with no additional donor risk factors, lower risk recipients, and short cold/warm ischemia.     

Continuous infusion of nitroglycerin improves pulmonary graft function of non-heart-beating donor lungs

BACKGROUND: The warm ischemic period of lungs harvested from a non-heart-beating donor (NHBD) results in an increased ischemia-reperfusion injury after transplantation. The intravenous application of nitroglycerin (NTG), a nitric oxide (NO) donor, proved to be beneficial during reperfusion of lung grafts from heart-beating donors. The objective of the present study was to investigate the effect of nitroglycerin on ischemia-reperfusion injury after transplantation of long-term preserved NHBD-lungs. METHODS: Sixteen pigs (body weight, 20-30 kg) underwent left lung transplantation. In the control group (n=5), lungs were flushed (Perfadex, 60 mL/kg) and harvested immediately after cardiac arrest. In the NHBD group (n=5) and the NHBD-NTG group (n=6), lungs were flushed 90 min (warm ischemia) after cardiac arrest. After a total ischemia time of 19 hr, lungs were reperfused and graft function was observed for 5 hr. Recipient animals in the NHBD-NTG group received 2 microg/kg/min of NTG administered intravenously during the observation period starting 5 min before reperfusion. Tissue specimens and bronchoalveolar lavage fluid (BALF) were obtained at the end of the observation period. RESULTS: Compared with the control group, pulmonary gas exchange was significantly impaired in the NHBD group, whereas graft function in the NHBD-NTG group did not change. Leukocyte fraction and protein concentration in the BALF and histologic alteration of the NHBD-NTG group were not different from controls. CONCLUSIONS: Continuous infusion of NTG in the early reperfusion period improves pulmonary graft function of NHBD lungs after long-term preservation. The administration of an NO donor during reperfusion may favor the use of NHBD lungs to alleviate the critical organ shortage in lung transplantation.     

Pre-arrest heparinization and ventilation during warm ischemia preserves lung function in non-heart-beating donors

BACKGROUND/PURPOSE: To solve the problem of donor scarcity, many attempts have been made including improved community education, relaxed organ acceptance criteria, increased reliance on single lung transplantation, and the use of partial organ donation. Unfortunately, these efforts have produced only modest increases in lung allograft availability; therefore, the so-called non-heart-beating organ donation must be considered. The aim of this study is to assess the viability of the non-heart-beating donor (NHBD) lung transplant rat model and determine the best strategy to manage the donor before and after cardiac arrest. METHODS: Fifty-five inbred Fischer rats were used as donors and recipients in an isogenic model of left lung transplantation. The rats were divided into 6 groups (n = 5): group I, normal controls without transplant; group II, heart-beating donor controls (HBD); group III, NHBD, no heparin, no ventilation during warm ischemia; group IV, NHBD, heparin, no ventilation; group V, NHBD, no heparin, ventilation; group VI, NHBD, heparin, ventilation. All lungs were stored at 4 degrees C for 4 hours. Animals were killed 24 hours after implantation. Gas exchange, pulmonary artery pressure, compliance, chest x-ray score, and histological score were assessed. RESULTS: Heparinized and ventilated animals during warm ischemia (group VI) had similar performance than those transplanted without warm ischemia time in a scenario of heart-beating donor (group II). Groups III, IV, and V transplanted lungs showed severe damage. CONCLUSIONS: The authors conclude that the rat lung transplantation model is useful to study the phenomena that occur in a setting of transplantation using NHBD and that heparinization and ventilation before cardiac arrest is the best strategy to manage non-heart-beating donors in this model.     

31P NMR assessment of orthotopic rat liver transplant viability. The effect of warm ischemia.

The relationship between NMR visible high energy phosphates and transplant outcome for the case of liver damage by warm ischemia was investigated. In vivo 31P nuclear magnetic resonance (NMR) spectroscopy of rat liver was performed before the induction of warm ischemia in the donor and 20 min after reestablishment of portal blood flow in the recipient. Pretransplant damage was varied by subjecting the livers to 0, 15, 30, or 60 min of warm ischemia prior to harvesting. In the controls (0 min warm ischemia), 4 of 4 rats survived transplantation (one week survival end-point) and the mean NTP recovery was 94 +/- 8%; 3 of 6 rats survived in the 15 min warm ischemia group. Mean NTP recovery was 77 +/- 20% in the 15 min survival subgroup and 32 +/- 20% in the nonsurvival subgroup. Of 6 rats, 1 survived in the 30 min group. NTP recovery was 44% for the 30 min survivor and 37 +/- 5% in the nonsurvival subgroup. Of 4 rats, 1 survived in the 60 min warm ischemia group. NTP recovery was 56% for the 60 min survivor and 28 +/- 7% in the nonsurvival subgroup. Overall, there was a significant difference between the mean NTP recovery of the survival and nonsurvival subgroups (78 +/- 21% versus 31 +/- 18%, P < 0.001). The dividing line between the survival and nonsurvival groups was approximately 50% NTP recovery. Of 9 rats with liver NTP recovery greater than 50%, 8 survived while 10 of 11 rats with less than 50% recovery died. NMR visible NTP recovery 20 min after the reestablishment of portal blood flow was a good indicator of transplant outcome in the case of rat liver damage by warm ischemia.