Celsior液、HTK液和UW液对心脏保存效果的实验研究

目的应用Langendorff灌注模型低温灌注离体鼠心6h,检验器官保存液Celsior液、HTK液和UW液对心脏保存的效果.方法实验分为3组,每组8只Wistar大白鼠.麻醉并抗凝后,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.分别用三种器官保存液灌注离体鼠心,在4℃下低温浸泡保存6h.重新复温、复灌,再次测定血流动力学值和冠状动脉流量,留取标本分别测定心肌水含量、心肌酶漏出量、心肌细胞ATP和CP含量和观察心肌细胞超微结构变化.结果①血流动力学恢复率,Celsior液和HTK液组均优于UW液组.②心肌酶漏出量,Celsior液和HTK液组明显低于UW液组.③ATP和CP含量,Celsior液和HTK液组高于UW液组.④超微结构变化,Celsior液组和HTK液组心肌损害最轻,UW液组心肌损害较重.结论Celsior液、HTK液和UW液对心脏保护的效果均较好,均为较合适的心肌保存液.Celsior液和HTK液对心脏保存的效果无显著差异,但均优于UW液.     

新型肾脏保存液低温保存后犬肾移植效果评价

目的建立犬自体肾移植模型,评价经新型肾脏保存液（HCA-Ⅱ）低温保存后的犬肾移植效果。方法60只杂种犬随机分为A（H-CA液,保存48h）、B（HCA—I液,保存48h）、C（UW液,保存48h）、D（H-CA液,保存72h）、E（HCA-Ⅱ液,保存72h）、F（UW液,保存72h）等六组。切取左肾,灌洗并放入该保存液保存。左肾动脉与右侧髂外动脉端端吻合,静脉与右侧髂外静脉端侧吻合,输尿管与膀胱抗反流吻合,同时切除右侧肾脏。检测移植前后肾功能指标。结果手术成功率95％,HCA-Ⅱ液各组术后血肌酐的峰值均低于H-CA液组,且血肌酐恢复正常的时间也较H—CA液组早（P〈0．05）;以上各指标均与UW液组无明显差别（P〈0．05）。结论HCA-Ⅱ肾脏保存液对供肾的保存效果明显好于H—CA保存液,可安全保存犬肾72h。     

常用心脏保存液及其改良方法

心脏移植是治疗终末期心力衰竭的最佳方案,但由于捐献器官的不足,心脏移植手术的开展受到极大限制。心脏保存液的改良对于扩大移植心脏供体池极为关键,该文介绍UW液、HTK液和Celsior液等3种常见的心脏保存液及其改良方法。     

SMO保存液对犬肾低温保存期间氧自由基变化的研究

建立犬离体肾脏单纯低温保存模型,对照组供肾灌注和保存用HTK保存液和UW保存液,实验组用自制SMO保存液,分别在犬肾保存24、48、72h切取肾皮质标本,测定皮质丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性。结果显示,实验组SOD活性48h明显高于对照组,MDA含量各时点均显著低于对照组（P均〈0．05）。实验组各时点SOD活性和MDA含量与UW液组比较均无统计学差异。提示SMO液在减轻氧自由基损伤方面优于HTK液,与UW液相当。     

左旋泊尼汀对离体大鼠心脏保存效果的影响

将25只成年雄性Wistar大鼠（体重350～400g）随机分为A组（对照组）及B、C、D、E组,每组5只,经麻醉和抗凝后,快速取下心脏。A组心脏悬挂在Langendorff灌注模型上K-H液恒压灌注,37℃下测定血流动力学指标;B、C、D、E组分别经主动脉灌注4℃ HTK液、UW液、HTK液＋左旋泊尼汀、UW液＋左旋泊尼汀心肌保存液,置于4℃相应的心肌保存液中保存6h;重新复温复灌。再次测定血流动力学指标,留取标本分别测定心肌含水量、心肌酶漏出量、ATP含量变化。结果 D组及E组平均左室发展压（LVDP）与B、C组比较显著升高,磷酸肌酸激酶同工酶（CK—MB）漏出量明显减少,心肌ATP含量明显增多。认为HTK液和UW液中加入左旋泊尼汀能改善离体鼠心脏的低温保存效果。     

UW器官保存液对供心保护的研究进展

UW器官保存液的特点是高钾、低钠、无钙，主要胶体成分是羟乙基淀粉、蜜三糖及乳糖醛酸。本文将介绍UW器官保存液近年来的研究进展。     

供体猪心在UW液中的保存时间和温度对其组织结构的影响

目的:探究体外保存时间和UW液温度对供体猪心组织结构的影响.方法:按照临床上现行的供体心脏标准获取流程获取18头中华小型猪的心脏,分别用21℃、4℃和-4℃的UW液在体外保存供体猪心;每一温度下,分别保存供体猪心3 h、6 h和9 h(每种保存条件下保存2个猪心).然后对体外保存后的猪心进行组织取材,通过苏木素-伊红(...     

自制AMU液对低温保存兔肾氧自由基表达的影响

目的观察自制AMU液对低温保存兔肾氧自由基表达的影响。方法分别采用HCA液、AMU液保存兔肾（HCA组和AMU组）。于保存24、48、72 h时切取肾皮质,分别用硫代巴比妥酸法（TBA法）和黄嘌呤氧化酶法检测肾皮质丙二醛（MDA）含量和超氧化物歧化酶（SOD）活力。结果随保存时间延长,两组SOD活力均下降、MDA含量均上升,但保存48、72 h时AMU组SOD活力明显高于HCA组（P均〈0.05）,保存各时点AMU组MDA含量低于HCA组（P均〈0.05）。结论在阻止低温保存兔肾SOD活力下降及MDA含量上升方面,AMU液优于HCA液。     

零下非结冰UW液、Celsior液和HTK液保存C3A细胞的效果比较

目的 比较UW液、Celsior液和HTK液零下非结冰(-0.8℃)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-0.8℃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)、LDH释放、尿素合成功能及白蛋白分泌功能.结果 UW液及Celsior液比HTK液显著提高了零下非结冰保存72 h的C3A细胞的存活率[(84.34±4.25)%,(83.53±3.73)% vs (75.65±3.01)%,P<0.001]和细胞内ATP含量[(5.54±1.44)μg/106 个细胞,(5.51±1.31)μg/106个细胞 vs (2.75±1.03)μg/106 个细胞,P<0.001];抑制了LDH释放(P<0.001);更好地维持了细胞尿素合成功能[(0.63±0.10)mmol/L,(0.62±0.06)mmol/L vs (0.39±0.04)mmol/L,P<0.001]和白蛋白分泌功能[(1.99±0.38)g/L,(1.96±0.24)g/L vs (1.50±0.18)g/L,P<0.05].UW液与Celsior液零下非结冰保存C3A细胞的效果无差异.结论 同HTK液相比,使用UW液或者Celsior液零下非结冰(-0.8℃)保存C3A细胞可以明显地提高复温后细胞存活率和细胞内ATP含量,降低低温损伤引起的LDH释放,有效地保护肝细胞尿素合成功能和白蛋白分泌功能.UW液同Celsior液零下非结冰保存C3A细胞的效果无差异,保存时间不宜超过72 h.     

新型心脏保存液研究进展

静态冷藏保存液保存心脏的临床效果表现良好,但其仅4-6小时的安全保存时限局限了供体的选择范围,限制了心脏移植手术的广泛开展。因此改良保存液成分,提升其性能延长保存时间以获得更好的保护效果势在必行。本文就心脏保存液的临床现状以及新型心脏保存液的研究进展作一综述。     

UW solution: a promising tool for cryopreservation of primarily isolated rat hepatocytes

Cryopreserved hepatocytes are a ready source of metabolic and synthetic functions for hepatocyte transplantation and bioartificial livers. In this study, we evaluated a cytoprotective effect of University of Wisconsin (UW) solution during cryopreservation of rat hepatocytes. We also investigated the feasibility of lentivirus‐based gene transfer into thawed hepatocytes after cryopreservation. Primary rat hepatocytes were isolated using a two‐step collagenase perfusion technique, and the resulting hepatocytes with more than 85% viability assessed by a trypan blue exclusion test were subjected to the present study. These cells were subjected to the present study. Cells were cryopreserved with UW solution containing 10% fetal bovine serum (FBS) with 12% dimethylsulfoxide (DMSO) (group 1, G1), Cellbanker solution (group 2, G2), and 10% FBS‐containing Dulbecco modified Eagle medium (DMEM) with 12% DMSO (group 3, G3). After thawing the cryopreserved hepatocytes, cell viability, plating efficiency, morphological appearance, and ammonia clearance activity were determined for each group. The efficacy of lentivirus‐mediated Escherichia coli LacZ gene delivery was evaluated. Hepatocyte viabilities after 3‐ and 7‐day cryopreservation were 73.2% and 62.5% for G1, 57.5% and 46.5% for G2, and 57.3% and 41.5% for G3, respectively. Plating efficiency and ammonia clearance activity were improved in G1 hepatocytes compared to G2 and G3 cells. Lentiviral transfer of a LacZ gene was confirmed in the thawed hepatocytes after cryopreservation by an X‐gal stain assay.     

不同保存液对静脉移植物的保护作用研究

目的 探究搭桥过程中不同保存液对血管的近期及远期保护作用.方法 健康新西兰大耳兔48只,分4组.A组：GV液（维拉帕米＋硝酸甘油）组,B组：肝素化全血组,C组：罂粟碱组及D组：对照组（林格液＋肝素）.取兔颈外静脉,分别放置于不同保存液中,30 min后移植至颈内动脉.于7 d、14 d、28 d取出移植物,HE染色观察不同时期内皮增厚及中膜增生情况,计算I/M值.同时取5只兔股静脉,粗细均匀,分为40段约0.5 cm长短静脉,分别放入1 ml不同保存液中,30 min后取出静脉,Elisa法测量1 ml保存液中内皮素（ET-1）含量,HE染色观察短期保存后内皮细胞破坏情况.结果 14 d及28 d时各组比较,A组及B组血管内皮破坏及内膜增厚最轻,C组及D组内皮破坏及增生最为严重.第2周及第4周时A组及B组I/M值比较差异无统计学意义,与C组及D组比较差异有统计学意义（P＜0.05）.Elisa法测定ET-1含量,A组释放量较B组、C组、D组均明显降低（P＜0.01）,A组及B组短期保存后的血管内皮破坏情况较C组、D组要轻,内皮细胞覆盖率较高.结论 GV保存液及肝素化全血保存液可以更好地保护血管内膜,A组保存液较B、C、D组的ET-1释放量更少.     

The challenge of improving donor heart preservation

Heart transplantation has in recent years become the treatment of choice for end stage heart failure. However while the waiting list for transplantation is growing steadily, the donor pool is not increasing. Therefore, in order to meet demand, transplant programs are using older, "marginal donors" and accepting longer ischaemic times for their donor hearts. As donor organs are injured as a consequence of brain death, during the period of donor management, at organ harvest, preservation, implantation and reperfusion, expansion of acceptance criteria places a great burden on achieving optimal long-term outcomes. However, at each step in the process of transplantation strategies can be employed to reduce the injury suffered by the donor organs. In this review, we set out what steps can be taken to improve the quality of donor organs.     

University of Wisconsin solution versus modified collins solution for canine heart preservation: An experimental study

We investigated the efficacy of University of Wisconsin (UW) solution for its ability to preserve hearts for 12 hours as compared with the results with modified Collins (MC) solution. After orthotopic transplantation, hearts stored in the UW solution (68%) showed significantly (p<0.05) better recovery of maximum rate of increase of left ventricular pressure than those stored in the MC solution (41%) (MC group: n=6, UW group: n=7). The serum CPK values 2 hours after reperfusion were significantly (p<0.01) higher in the MC group (6528 IU/L) than in the UW group (2021 IU/L). Myocardial adenosine triphosphate (ATP) levels were measured using phosphorus 31 nuclear magnetic resonance spectroscopy during 12 hours of cold storage (MC group: n=5, UW group: n=4). ATP levels continued to decrease throughout the storage, and fell to 39% (MC group) and 23% (UW group) of the control after 12 hours with no significant difference. Significant differences between the two groups were present in water content before (MC group: 77.6%, UW group: 75.8%;p<0.05) and after (MC group: 77.6%, UW group: 75.8%;p<0.025) 12 hours of storage (MC group:n=6, UW group:n=5). These results demonstrated that the UW solution was superior to the MC solution when used to preserve hearts for 12 hours.     

一氧化碳气体干燥保存法对小鼠心脏保存效果的实验研究

目的观察一氧化碳（carbon monoxide,CO）气体干燥保存法对同种小鼠心脏异位移植的影响。方法采用C57BL／6雄性小鼠,建立同系小鼠心脏腹部异位移植模型。4—6周龄小鼠60只,分为3组（n=20）,即空白对照组,HTK供体心组及CO供体心组;8～10周龄小鼠40只,分为HTK受体组及CO受体组（n=20）。取HTK供体心组小鼠心脏浸泡于HTK液中;取CO组小鼠心脏悬挂并置于高压气体罐中（PO2：3．2×101．3kPa＋PCO：0．8×101．3kPa=4×101．3kPa）。两组供体心均置于4℃冰箱中保存8h后,分别移植于两受体组小鼠腹部。空白对照组仅取供体心,不做移植术。移植完成后,比较HTK组及CO组复跳时间。移植2h后,对比观察两组移植心脏的颜色及复跳情况。取3组供体心心肌组织,分别采用苏木素伊红染色和TUNEL染色法对比供体心病理学改变及心肌细胞的凋亡情况。结果HTK供体心组6只复跳,CO供体心组13只复跳,复跳率差异有统计学意义[86．7％（13／20）vs.53.3％（7／20）,P=0．027];与HTK供体心组相比,CO供体心组供体心复跳时间显著缩短,差异有统计学意义[（203．92±73．52）s vs．（352．67±61．19）s,P=0．001];病理方面,与空白对照组比较,CO供体心组供体心病理损伤明显,但与HTK组比较明显减轻;Tunel检测显示,CO组凋亡率较空白对照组升高,差异有统计学意义（P〈0．05）;但比HTK组明显减少,差异有统计学意义（P〈0．05）。结论CO气体保存法可以缩短供体小鼠心脏复跳时间及减少心肌细胞的凋亡,对离体心脏具有显著的保护作用。     

Efficacy of hypothermic perfusion using University of Wisconsin solution in extended hepatectomy with hepatic inflow occlusion in a canine model

This study was designed to elucidate the efficacy of University of Wisconsin (UW) solution for preventing liver injury, when used as a hypothermic perfusate infused into the systemic circulation during extended hepatectomy with hepatic inflow occlusion. Adult mongrel dogs (9.5–17.5 kg, n= 14) were subjected to 75% hepatectomy under 60 min hepatic inflow occlusion. The animals were divided into two groups. The UW group (n= 7) underwent hypothermic perfusion using 4°C UW solution (core temperature of the liver: 12.3±0.2°C). The control group designated as the Ringer's lactate (LR) group (n= 7) underwent hypothermic perfusion using 4°C LR solution. The perfusate was introduced into the systemic circulation via the hepatic vein. Blood from the hepatic vein was sampled, and alanine amino-transferase, purine nucleoside phosphorylase activities and the ammonia concentration were measured. The 7 day survival rate was higher in the UW group than in the LR group. The parameters of liver function were less significantly altered in the UW group than in the LR group. The plasma ammonia concentration was significantly (P<0.05) lower 6h after reperfusion in the UW group than in the LR group. A small volume of hypothermic perfusion of the liver using UW solution was safe if it returned to systemic circulation. Hypothermic perfusion of the liver using UW solution may be effective for preventing hepatic tissue injury during extended hepatectomy with hepatic vascular occlusion.     

Adenosine 5＇triphosphate transport and accumulation during the cold preservation of rat hepatocytes in University of Wisconsin solution

AIM: We used isolated hepatocytes to investigate how different concentrations of ATP in the University of Wisconsin (UW) solution affected both cellular ATP content and cell viability during the cold storage and the rewarming step. The mechanism involved in ATP transport and accumulation in hypothermia was also determined. METHODS: The cells were preserved up to 72 h in different conditions: UW solution without ATP (a-group), UW+5 mmol/L ATP (b-group), and UW+10 mmol/L ATP (c-group). The ATP content and the cell viability (LDH release) were determined during the cold storage and the rewarming step. In the groups a and c, the respiratory function of the cells at rewarming was studied. In addition, the cell volume of hepatocytes and the mechanism involved in ATP transport and accumulation were assessed. The extracellular degradation of exogenous nucleotides during transport experiments was investigated by a HPLC technique. RESULTS: After three days of cold storage a loss of cellular ATP content was observed in hepatocytes preserved either without nucleotides (a-group) or with 5 mmol/L ATP (b-group). In contrast, 10 mmol/L ATP (c-group) was able to maintain a normal ATP cellular content, with only a 6% diminution after 72 h of cold storage. The respiratory function was significantly different in hepatocytes preserved with 10 mmol/L ATP than without ATP. No significant change was detected For the three groups in cellular volume during the cold storage. We also report that the time course accumulation of [3H]-ATP by cold stored hepatocytes is a rapid process that is completed after 180 s with linear dependence on the extracellular ATP concentration (linear fitting results in a slope of 0.5624±0.1179 mmol/L ATP intracell/mmcl/L ATP extracell). CONCLUSION: Our results show that, during hypothermic storage in UW solution, hepatocytes are permeable to ATP by a diffusive mechanism. Also, we found that it is ATP the main extracellular nucleotide available for transport and it is not the breakdown products.     

缬沙坦对自发性高血压大鼠肾脏的保护作用

目的探讨缬沙坦对自发性高血压大鼠(SHR)肾脏的保护作用,以及与结缔组织生长因子(CTGF)在SHR肾脏中表达的关系。方法本实验以12周龄雄性SHR和同龄雄性WistarKyotoRats(WKY)大鼠为研究对象,SHR分为治疗组[缬沙坦,30mg/(kg·d)、灌胃]与对照组,WKY大鼠为正常对照组,测量不同时期(12周,24周)大鼠尾动脉压、肾功能、尿β2微球蛋白(β2MG)的变化情况,以及用免疫组化的方法观察结缔组织生长因子在肾脏中的表达。结果12周后,相对于同龄WKY大鼠、SHR治疗组而言,SHR对照组的尿素氮、血肌酐变化无显著意义,而尾动脉压、尿β2MG均有显著增高(P<0.01,P<0.05)。经免疫组化半定量检测显示:在24周龄SHR对照组,其肾脏中CTGF的表达相对于同龄WKY大鼠及SHR治疗组明显升高(P<0.01),同时与尿β2MG的水平成正比(P<0.05)。结论缬沙坦能显著减少尿蛋白,同时使CTGF在SHR肾脏的表达明显减少,说明CTGF可能是慢性高血压导致的肾损害的重要机制之一。     

葡萄糖-胰岛素-钾液对缺血/再灌注心肌的保护作用

目的:观察极化液（葡萄糖-胰岛素-钾液,G IK）对急性心肌缺血/再灌注（M I/R）犬心脏功能、冠脉血流量及心肌损伤的影响,分析胰岛素在G IK上述效应中的作用。方法:制备犬M I/R模型,心肌定量缺血（左前降支血流量降低80%）50 m in,再灌注4 h。24只杂种犬随机分为G IK、葡萄糖-钾液（GK）和盐水对照组（n=8/组）,再灌注前5m in输注G IK、GK、生理盐水。观察冠脉血流量及血流动力学指标;检测不同时间血清乳酸脱氢酶（LDH）、肌酸激酶（CK）活性;再灌注4 h后测量并计算心肌梗死范围。结果:与盐水对照组相比,G IK明显增加左前降支冠脉血流量（CBFLAD）,改善再灌注后左室收缩及舒张功能,降低血清CK、LDH,减少心肌梗死范围,而GK无上述作用。结论:再灌注时输注G IK可促进再灌注心脏功能恢复及减轻心肌损伤,该作用可能与G IK增加冠脉血流量有关;胰岛素是G IK上述作用的关键成分。