离体鼠心低温器官保存液保存后超微结构的分析

目的评价阜外改良心肌保存液(fuwai mod ified solution,FWM),HTK液(h istid ine-tryptophan-ketogluratesolution)和UW液(un iversity of w isconsin solution)对离体鼠心的保存效果。方法20只雄性W istar大鼠随机分为四组(Control组、FWM组、HTK组和UW组),每组5只。Control组切取鼠心后立即移至Langendorff装置灌注1h,其它三组分别用不同的保存液4℃下保存6 h,然后将离体鼠心移至Langendorff装置复灌1 h,测定复灌1h后的心肌含水量变化,并用透射电镜观察复灌1 h后心肌超微结构的改变。结果UW组心肌含水量高于Control、FWM和HTK组,而Control,FWM和HTK三组相比无显著统计学差异(P>0.05)。电镜结果显示,Control组和FWM组心肌超微结构基本正常,HTK组出现心肌超微结构改变,而UW组心肌超微结构改变严重。结论FWM液的心肌保存效果优于HTK和UW液。     

TRS液低温下长时间保存血管的效果

目的观察检测一种新型保存液（tongji rochester solution，TRS）在4℃下保存24h后对大鼠主动脉保护作用。方法用大鼠胸主动脉分别保存在4℃ TRS液、UW液、HTK液、生理盐水中保存24h，在甲氧胺和乙酰胆碱作用下分别检测其血管平滑肌细胞的收缩功能和舒张功能。未经保存的新鲜血管作为对照。结果保存24h后，血管平滑肌细胞的收缩功能和内皮依赖性舒张功能在TRS液、UW液、HTK液组之间差异无显著性（P〉0．05）。结论TRS在4℃、24h内对血管的保存效果与UW液、HTK液相当，为一种理想的保存液。     

上海多器官保存液保存离体大鼠肝脏的实验研究

目的:观察上海多器官保存液(Shanghai-mutil-organ solution,SMO液)对离体大鼠肝脏的保存效果,探讨应用SMO液保存离体供肝的可行性.方法:SD大鼠随机分为SMO液、UW液和HTK液保存组,建立离体肝脏单纯低温保存模型,保存液保存8、16、24、36 h分析肝脏组织能量代谢情况,观察肝组织形态学改变和肝脏细胞凋亡情况.结果:保存16、24、36 h,SMO液组肝组织三磷酸腺苷(ATP)、磷酸腺苷总量(TAN)及Atkinson能荷(AEC)均明显高于同时点HTK液组 (P＜0.05),与同时点UW液组无显著差异;形态学检查见SMO液组组织损伤较同时点HTK液组轻,除细胞肿胀较同时间点UW液组明显外,其余表现基本一致.保存24、36 h,SMO液组凋亡指数明显低于HTK液组(P＜0.05),而与UW液组无明显差异.结论:SMO液对大鼠离体肝脏的保存效果总体上与UW液相当,优于HTK液,仅在防止细胞水肿方面较UW液稍差.     

尼可地尔改善心脏停搏液对心脏的保护作用

目的　观察在HTK液和UW液中加入尼可地尔 (Nicorandil)对心肌保存液保存效果的影响。方法　实验分为 4组 :UW液 (U)组 ;HTK液 (H)组 ;UW液组 +Nicorandil(U +N)组 ;HTK液 +Nicorandil(H +N)组 ;每组 8只大白鼠。麻醉和抗凝后 ,快速取下鼠心并悬挂在Langendorff灌注模型上灌注 ,测定血流动力学基础值。分别用四组保存液灌停离体鼠心并低温 (4℃ )浸泡保存 6h。复温、复灌后再次测定血流动力学值 ,留取标本分别测定心肌水含量、心肌酶漏出量、心肌细胞ATP和CP含量和心肌细胞超微结构变化。结果　心肌血流动力学恢复率 :H +N组>U +N组 >H组 >U组。心肌水含量 :4组间差异无显著意义。灌脉流出液中心肌酶漏出量 :H +N组 U +N组 >H组 >U组。心肌细胞超微结构变化 :H +N组心肌损害最轻 ,H组轻于U +N组 ,U组心肌损害最严重。结论　HTK液和UW液都是比较合适的心肌保存液 ,HTK液对心肌的保护作用优于UW液 ,钾离子通道开放剂Nicorandil能够善器官保存液HTK液和UW液对心肌的保存效果。     

生脉散对大鼠离体肝脏保护作用的实验研究

目的:研究生脉散对大鼠离体肝脏的保存效果。方法:采用大鼠肝脏非循环离体灌注模型(noncirculated isolatedperfusion of rat liver),以生理盐水、生脉散、UW液(Universiev of Wisconsin)、生脉散联合UW液对大鼠离体肝脏保存。测定其8、16、24和48h灌注流出液中天冬氨酸转氨酶(AST)、氧自由基代谢产物丙二醛(MDA)的含量,以及保存8小时组织病理切片。通过生脉散组与生理盐水组比较和生脉散联合UW液与UW液相比较,从而了解生脉散对大鼠肝脏有无保护作用。结果:生脉散组各时相流出液内天冬氨酸转氨酶(AST)比生理盐水组明显低,有显著差异(P<0.05),MDA含量低于生理盐水组,有显著差异(P<0.05)。生脉散联合UW液灌注流出液各时相AST含量与UW液保存者相近,无显著差异(P>0.05),生脉散联合UW液灌注流出液MDA含量低于UW液组,有显著差异(P<0.05)。光镜下观察形态学,生脉散组优于生理盐水组,生脉散加UW液组与UW液组变化基本一致。结论:生脉散液对大鼠肝脏保存效果在MDA、AST都好于生理盐水组,与UW液联合运用MDA方面优UW液,在AST方面无明显差异。     

UW液、Celsior液和HTK液低温保存生物人工肝用L-02细胞的效果比较

目的 比较UW液、Celsior液和HTK液4℃常规低温保存生物人工肝用L-02细胞的效果。方法 制备好的L-02细胞悬液分以下3组：UW液保存组（UW液组）;Celsior液保存组（CS液组）;HTK液保存组（HTK液组）。各组细胞于4℃低温保存72h后,分别测定细胞存活率及死亡率（流式细胞术）,ALT及LDH释放,尿素合成功能及白蛋白分泌功能。结果 UW液较Celsior液和HTK液显著提高了4℃常规低温保存保存72h的L-02细胞的存活率[(60.05 ± 4.23)% vs (50.12 ± 3.99)%、(44.20 ±4.67)%],均P<0.05;降低了细胞死亡率[(39.95±4.23)%vs (49.88 ±3.99)%、(55.80%±4.67)%], 均P<0.05;抑制了ALT、LDH释放(均P <0.05);更好地维持了L-02细胞尿素合成功能[ (1.03 ± 0.23)mmol/L vs (0.80± 0.14)mmol/L、(0.48± 0.05)mmol/L]和白蛋白分泌功能[(8.36 ±1.38 )mg/L vs (6.41±1.25)mg/L、(5.19±0.41)mg/L), 均P<0.05。结论 同Celsior液和HTK液相比,使用UW液4℃常规低温保存L-02肝细胞可以明显的提高复温后细胞存活率,降低低温损伤引起的ALT、LDH释放,有效的保护肝细胞尿素合成功能和白蛋白分泌功能,但保存时间不应超过72h。     

三种肝保存液低温保存生物人工肝用C3A细胞的效果比较

目的比较UW液、Celsior液和HTK液4℃常规低温保存生物人工肝用C3A细胞的效果。方法制备好的C3A细胞悬液分以下3组:UW液保存组(UW液组);Celsior液保存组(CS液组);HTK液保存组(HTK液组)。各组细胞于4℃低温保存72 h后,分别测定细胞存活率及死亡率(流式细胞术),丙氨酸氨基转移酶(ALT)及乳酸释放,尿素合成功能及清蛋白分泌功能。结果 UW液及Celsior液比HTK液显著提高了4℃低温保存72 h的C3A细胞的存活率(78.56%±4.67%vs 76.03%±3.53%,60.54%±3.18%,P<0.05);抑制了C3A细胞低温保存时ALT释放及乳酸的释放(P<0.05);更好地维持C3A细胞尿素合成功能[(0.52±0.11)mmol/L vs(0.51±0.06)mmol/L,(0.32±0.05)mmol/L,P<0.05]和清蛋白分泌功能[(1.79±0.26)g/L vs(1.75±0.21)g/L,(1.20±0.17)g/L,P<0.05]。UW液同Celsior液4℃低温保存C3A细胞的效果无差异。结论同HTK液相比,使用UW液或者Celsior液4℃保存C3A细胞可以明显的提高复温后细胞存活率,降低低温损伤引起的ALT释放和乳酸释放,有效的保护肝细胞尿素合成功能和清蛋白分泌功能。     

深低温下阜外改良器官保护液对大鼠胸主动脉内皮源性超极化因子介导的舒张功能的影响

目的探讨深低温下阜外改良（Fu Wai Modified，FWM）器官保护液对离体大鼠胸主动脉内皮源性超极化因子（EDHF）介导的舒张功能的影响。方法取大鼠胸主动脉，切成2mm的血管环，分别用FWM液、UW液、HTK液、Krebs-Henseleit重碳酸盐缓冲液（KH）液在4℃缺氧条件下保存4h后，采用器官槽法，检测在环加氧酶阻断剂indomethacin（7μmol／L）和一氧化氮合成酶阻断剂LNNA（300μmol／L）作用下，前列腺素F2α（U46619 30nmol／L）及非受体介导钙离子载体（A23187—6log mol／1）引发的收缩舒张反应。结果A23187引发的舒张反应，FWM组高于KH组（P〈0．05），UW组低于KH组（P〈0．05），HTK组高于UW组（P〈0．01），FWM组与HTK组相比，差别无显著性（P〉0．05）。结论深低温下，FWM，HTK液对大鼠胸主动脉EDHF介导血管舒张功能有保护作用，UW液对其有抑制作用。     

SMO保存液对犬肾低温保存期间Na~+-K~+ ATPase的影响

目的:研究SMO保存液对犬肾低温保存期间Na+-K+ATPase的影响。方法:建立犬离体肾脏单纯低温保存模型,对照组用0~4℃HTK保存液和UW保存液对供肾进行灌注和保存,试验组用0~4℃自制SMO保存液对供肾进行灌注和保存,分别于24、48、72h后取皮质标本,进行组织病理学观察及检测Na+-K+ATPase的活性。结果:SMO液组所见组织病理学改变,48h和72h比HTK液组轻,在各时间点与UW液组基本相似。Na+-K+ATPase活性,保存各时点,SMO液组与UW液组无明显差异;保存24h和48h,SMO液组与HTK液组无明显差异;保存72h,SMO液组Na+-K+ATPase活性明显高于HTK液组,存在明显差异。结论:SMO保存液在减轻细胞形态学改变和减缓Na+-K+ATPase活性下降方面显著优于HTK保存液,与UW保存液相当。     

阜外极化停搏液大鼠离体心肌保护效果研究

目的研究阜外极化停搏（FWP）液对大鼠离体心肌保护的效果。方法SD大鼠24只,随机分为四组,对照组,STH-2液（St.ThomasⅡ）组,HTK液（histidine-trptophan-ketoglurate）组和FWP液（fuwai polarizing）组。除对照组,其他各组心脏均在4℃心肌保护液中保护1 h。观察左心室功能恢复情况、冠状动脉流量（CF）、冠状静脉引流液中肌酸激酶同工酶（CKMB）、乳酸脱氢酶（LDH）、肌钙蛋白I（cTnI）含量以及心肌细胞的超微结构等。结果各实验组心功能指标均有不同程度下降。FWP组和HTK组LVDP,CF和-dp/dtmax显著高于STH-2组（P〈0.01）;FWP组dp/dtmax显著高于HTK组和STH-2组（P〈0.01）;FWP组和HTK组心肌酶CKMB、LDH和cTnI含量显著低于STH-2组。心肌超微结构显示FWP组和HTK组超微结构保存较好,损伤比对照组小。结论FWP液组对心肌保护效果明显优于STH-2液组,与HTK液相似。     

The Effect of Fructose-1,6-diphosphate and HTK Solution on Protecting Primary Cardiac Muscle Cells of Rat with Cold Preservation

It is one of the sticking points for organtrans-plantation to protect the donated organs . With thedevelopment in the preservation solution for or-gans ,the survival rate of organ transplants hasbeen greatly i mproved. At present , heart trans-plantation has become an effective treat ment forend-stage heart diseases . The need for organs of-fered is increasing, which also requires investiga-ting and i mproving the preservation solution. Theexperi ment was based on cultured pri mary cardiacmusc…     

卡立泊来德对鼠移植供心的保护效果

目的 通过离体心脏灌流模型(Langendorff),评价卡立泊来德(Cariporide)对离体鼠移植供心的保护效果.方法 48只雄性Wistar大鼠随机分为四组.对照组两组:HTK(Histidine-Tryptophan-Ketoglurate Solution)8h、12h组,实验组(Cariporide 组)两组:HTK+Cariporide 20μmol/L,8h、12h组.切取鼠供心后即悬挂在Langendorff上灌注,测定血流动力学基础值.分别用两种器官保存液灌注离体鼠心后置于各自的冷心肌保护液(4℃)中保存8h、12h,恢复灌注后再次测定血流动力学指标和冠状动脉流量,留取标本分别测定心肌水含量、心肌酶漏出量、心肌细胞ATP含量,观察心肌细胞超微结构变化.结果 在冷缺血8h后,与对照组相比,实验组的平均左室发展压(LVDP)及心肌ATP含量均增高,磷酸肌酸激酶同工酶(CK-MB)漏出量减少,但差异无统计学意义.在冷缺血12h后,与HTK组相比,卡立泊来德组LVDP均显著升高[(98.37±16.34)mmHg、(43.21±14.33)mmHg,(P＜0.01)](1 mmHg=0.133kPa);CK-MB漏出量明显减少[(4.373±1.489)IU/L、(34.23±3.328)IU/L,(P＜0.01)];心肌ATP含量明显增多[(34.32±1.87)μg/g、(17.92±1.47)μg/g,(P＜0.01)].结论 Cariporide对离体鼠供心具有较好的保护效果,并可延长供心的低温保存时间.     

Comparative Study on the Effects of Self-made Liver Preservation Solution on Secretion of ICAM-1 and NO in Rats

In order to study the effect of self-made liver preservation solution on liver preservation by comparing with UW solution and HC-A solution, the self-made liver preservation solution （SM） and perfusion solution were prepared under the aseptic conditions. The isolated non-circulated perfusion rat liver model was established. According to the different preservation solutions, the rats were randomly divided into UW group, SM group and HC-A group. The three groups were divided into 6 subgroups according to the preservation duration （n=6 in each group）. The transferase in liver perfusion solution and intercellular adhesion molecule-1 （ICAM-1） and nitric oxide （NO） in liver tissues were determined at 2, 8 and 24 h respectively. The results showed that the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） had no significant difference between SM group and UW group, but significantly lower than in HC-A group. The levels of ICAM-1 and NO were increased simultaneously in SM group and UW group （P〉0.05）, but there was significant difference as compared with HC-A group （P〈0.05）. At the same time point, the level of ICAM-1 was higher in SM group than in UW group, but NO was lower. The preservation effect of SM solution is the same as UW solution, but better than HC-A solution.     

肝移植3种器官保存液的比较研究

目的 比较UW液、Celsior液和HTK液时移植供肝灌注保存的效果.方法 回顾性分析2002年1月～2004年3月施行的成人首次肝脏移植且术前诊断为良性肝病者263例,根据所采用的灌注保存液分为lAW液组(161例)、Celsior组(45例)和HTK组(57例),3组患者年龄、性别及原发病分布,术前肝功能指标及分级,肝移植术式及供肝冷,热缺血时间差异均无显著性,比较3组患者术后早期移植肝功能指标、血管及胆道并发症发生率和移植物存活率.结果 3组患者肝移植术后早期血清丙氨酸氨基转移酶、门冬氨酸氨基转移酶、碱性磷酸酶和γ-谷氨酰转肽酶差异均无显著性;UW液组术后第7天血清总胆红素(117.4±96.3)μmol/L高于Celsior组[(85.1±77.9)μmol/L,P=0.047]和HTK组[(86.5±82.6)μmol/L,P=0.033];HTK组术后即刻凝血酶原时间(21.9±5.8)s低于UW液组[(24.8±6.2)s,P=0.004]和Celsior组[(26.4±7.0)s,P=0.0011;HTK组凝血酶原时问峰值(24.0±5.2)s低于Celsior组[(27.5±6.1)s,P=0.002].UW液组发生移植肝原发无功能2例(1.2%),但组间差异无显著性(P=0.528).3组患者术后血管并发症、胆道并发症及排斥反应发生率和再次移植率差异均无显著性.移植物1年及3年存活率分别为:UW液组80.7%和73.9%,Celsior组91.1%和82.2%,HTK组94.7%和80.7%,存活时间差异无显著性(P=0.099).结论 UW液、Celsior液和HTK液对移植供肝灌注保存的效果差异无显著性.     

Comparative Study on the Effects of Serf-made Liver Preservation Solution on Secretion of ICAM-1 and NO in Rats

In order to study the effect of self-made liver preservation solution on liver preservation by comparing with UW solution and HC-A solution, the self-made liver preservation solution (SM) and perfusion solution were prepared under the aseptic conditions. The isolated non-circulated perfu-sion rat liver model was established. According to the different preservation solutions, the rats were randomly divided into UW group, SM group and HC-A group. The three groups were divided into 6subgroups according to the preservation duration (n=6 in each group). The transferase in liver perfu-sion solution and intercellular adhesion molecule-1 (ICAM-1) and nitric oxide (NO) in liver tissues were determined at 2, 8 and 24 h respectively. The results showed that the levels of alanine aml- notransferase (ALT) and aspartate aminotransferase (AST) had no significant difference between SM group and UW group, but significantly lower than in HC-A group. The levels of ICAM-1 and NO were increased simultaneously in SM group and UW group (P＞0.05), but there was significant dif-ference as compared with HC-A group (P＜0.05). At the same time point, the level of ICAM-1 was higher in SM group than in UW group, but NO was lower. The preservation effect of SM solution is the same as UW solution, but better than HC-A solution.     

托马斯液复合应用康斯特保护液对大鼠离体心肌保护作用的研究

目的：观察托马斯液（STH）复合应用不同剂量康斯特保护液（HTK）对大鼠长时间缺血心肌的保护作用。方法：雌性Wistar大鼠40只,体重230～280g,随机分为S、H、S＋H10、S＋H20和S＋H30共5组（n=8）。麻醉后开胸取出心脏立即连于langendorff灌注装置,制备心脏缺血再灌注模型,S组用STH液15ml／kg每30min灌注1次。H组用HTK液30ml／kg单次灌注,其余3组均先用STH液15ml／kg灌注后分别续灌HTK液10ml／kg、20ml／kg、30ml／kg。分别留取停搏前、再灌注15、30min时冠脉流出液2ml用于检测心肌乳酸脱氢酶（LDH）、肌酸激酶（CK）。实验末留取心尖部心肌组织,用于检测心肌组织三磷酸腺苷（ATP）、丙二醛（MDA）、超氧化物歧化酶（SOD）含量。结果：与S组比较,其余4组复灌15、30min后冠状动脉流出液心肌酶CK、LDH含量较低（P〈0．05,P〈0．01）;与S、H、S＋H10各组相比,复灌30min后,S＋H20组及S＋H30组心肌ATP、SOD含量增高（P〈0．05,P〈0．01）,心肌MDA含量降低（P〈0．05,P〈0．01）。结论：STH液复合一定剂量的HTK液诱导并维持心脏停搏能增加大鼠离体心肌ATP、SOD含量,降低MDA含量．减轻缺血再灌注损伤。     

Cold preservation of pig liver grafts with warm ischemia and pentoxifylline-UW solution

BACKGROUND: We undertook this study to investigate the safe time limits of cold preservation in UW solution of liver grafts subjected to warm ischemia (WI) for 20 min and the changes of the limits when pentoxifylline is added to UW solution. METHODS: The safe time limit was studied in a simple porcine orthotopic liver transplantation (LTx) model. In donors, livers were subjected to 20 min of WI and subsequent 12-h (group 1, n = 5), 16-h (group 2, n = 5), and 20-h (group 3, n = 3) cold preservation in UW solution, respectively. After the safe time limits were clear, another group (group 4, n = 5) was built to test whether or not the limits can be changed when pentoxifylline is added to UW solution in an unsafe time limit group. RESULTS: All five animals in group 1 survived up to 7 days of the survey endpoint. In group 2, only one animal survived up to the same survey endpoint and all animals in group 3 died within 12 h. The 1-week survival rate of group 1 was significantly higher than the other two groups. Group 1 had a lower level of alanine aminotransferase (ALT) or aspartase aminotransferase (AST) after LTx, less pathological damage, higher concentration of adenosine triphosphate (ATP) and higher microcirculation blood flux in the grafted liver tissue at 1 h after reperfusion than the other two groups. The results primarily showed that 12-h cold preservation was safe, 16 h was unsafe, and 20 h was highly unsafe. But when pentoxifylline was added to UW solution in cold preservation (16-h group, group 4), in contrast to group 2, the incidence of liver tissue necrosis and primary graft nonfunction was significantly lower in group 4 than in group 2. The 1-week survival rate of the pigs was 100% in the former and 20% in latter group. Levels of ALT and AST in recipients' artery blood, malondialdehyde and TNF-alpha concentration in grafted liver tissue, resistance of portal vein and hepatic artery after preservation in group 4 were significantly reduced, whereas microcirculation blood flux of the grafted liver, superoxide dismutase concentration and ATP concentration in grafted liver tissue were significantly elevated. CONCLUSIONS: The safe time limit of cold preservation in UW solution of liver grafts subjected to WI for 20 min was about 12 h and the limits can be prolonged to 16 h when pentoxifylline is added to UW solution. Many mechanisms were involved.