肝素对大鼠减体积肝移植术后肝再生的影响
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摘要
【目的】供肝短缺严重地制约着临床肝移植的发展。随着活体肝移植技术的不断成熟,其已成为缓解供体短缺的重要途径之一。临床活体肝移植术后早期常应用肝素、低分子肝素钠或低分子肝素钙来预防或治疗肝动脉血栓形成,而肝素对活体肝移植术后移植肝脏和供体残余肝脏再生的影响缺乏相关研究。本实验在大鼠减体积肝移植模型的基础上,初步探讨肝素对肝再生的影响。
【方法】实验前期进行显微外科训练,尝试二袖套法建立大鼠全肝肝移植模型和系列减体积肝移植(60%、50%、30%)模型。在此基础上选取同季节健康雄性wistar大鼠120只(北京军事医学科学院实验动物中心),体重230-280g,随机分为四组(随机数表法):A、B两组每组42只,供受体随机选取,C、D两组每组18只。A、B两组行50%减体积肝移植术后分别给予生理盐水(4ml/kg)、肝素(1000IU/kg),C组行50%肝切除术后给予生理盐水(4ml/kg),D组为假手术组,术后给予生理盐水(剂量同C组),各组均为术后20h皮下给药。各组大鼠术后3天、5天、7天随机牺牲6只。牺牲鼠麻醉开腹后下腔静脉取血5ml,静置后离心取血清,酶联免疫吸附试验检测肝细胞生长因子(HGF);取移植肝脏,记录牺牲前大鼠重量、肝脏湿重,计算肝再生速率,取部分肝脏制作肝脏单细胞悬液检测肝细胞增殖指数,部分肝脏-80℃深低温保存,剩余肝脏甲醛固定,做常规HE染色和免疫组化染色(肝细胞增殖核抗原PCNA),记录PCNA表达情况。统计学分析比较A与B组、A与C组肝脏再生指标的差异(SPSS13.0,独立样本t检验),比较A与B组、A与C组HGF水平的差异。
【结果】前期完成一百余例大鼠全肝肝移植和系列减体积肝移植,建立了稳定的大鼠肝移植模型。A、B两组大鼠50%减体积肝移植术中供体切取时间分别为30±3min、31±4min,无肝期分别为15±2min、16±2min,冷缺血时间分别为56±5min、54±7min,受体手术时间分别为54±7min、58±5min。A、B两组术后48h存活率分别为95.2%(20/21)、100%(21/21),移植鼠无出血和感染死亡,A组两例因术后发生胆道并发症排除,胆道并发症发生率为10%(2/20),B组3例因术后发生胆道并发症排除,胆道并发症发生率14.3%(3/21)。C组完成大鼠50%肝切除18例,手术用时(14±2)min。C、D两组无死亡和并发症病例。A、B、C三组术后3天、5天、7天肝再生速率分别为(33.5±6.6)%、(43.7±5.4)%、(45.6±6.3)%,(40.1±2.8)%、(71.9±17.8)%、(56.9±5.4)%,(86.6±11.9)%、(111±19.2)%、(98.4±4.6)%。A、B两组术后3天、5天、7天肝再生速率比较均有统计学差异(P=0.015＜0.05、P=0.002＜0.01、P=0.023＜0.05)。A、C两组术后3天、5天、7天肝再生速率比较均有统计学差异(P=0.012＜0.05、P=0.011＜0.05、P=0.031＜0.05)。A、B、C组大鼠术后3天、5天、7天血清HGF水平分别为(263.3±101.1)pg/ml、(420.7±56.8)pg/ml、(365.3±6.4)pg/ml,(312.9±10.1)pg/ml、(440.9±23.4)pg/ml、(384.4±32.2)pg/ml,(242.4±22.1)pg/ml、(409.9±23.0)pg/ml、(376.3±22.3)pg/ml。A、B两组大鼠术后3天、5天、7天血清HGF水平比较有统计学差异(P=0.008＜0.01、P=0.000＜0.01、P=0.000＜0.01)。A、C两组术后3、5、7天血清HGF水平比较均有统计学差异(P=0.016＜0.05、P=0.038＜0.05、P=0.001＜0.01)。A、B、C组术后3天、5天、7天肝细胞增殖指数PI分别为:(30.4±4)%、(48.1±8)%、(38.4±2)%,(29.3±3)%、(43.5±8)%、(36.9±2)%,(30.3±3)%、(47.3±8)%、(31.6±3)%。A、B两组不同时间段PI指数水平比较均有统计学差异(P=0.001＜0.01、P=0.002＜0.01、P=0.02＜0.05)。A、C两组术后3、5天PI指数比较均有统计学差异(P=0.025＜0.05、P=0.018＜0.05),A、C两组术后7天PI指数比较无统计学差异(P=0.86＞0.05)。A、B两组术后3天、5天PCNA表达均有统计学差异,而术后7天PCNA表达无统计学差异。A、C两组术后3天、7天PCNA表达无统计学差异,术后5天PCNA表达有统计学差异。
【结论】皮下给予肝素(1000IU/kg)可促进大鼠50%减体积肝移植术后肝再生;大鼠减体积肝移植术后肝脏再生速率较肝切除术后肝再生延缓;本实验肝素促进血清HGF水平的升高可能是肝素促进肝再生的机制之一。
【Objective】Unfractionated heparin,low molecular weight heparin were regularly utilized to prevent or treat hepatic artery thrombosis during early postoperative in clinical living donor liver transplantation.However,there is few literatures on effects of heparin on hepatic regeneration after 50%partial liver transplantation in rats.This study was designed to investigate effects of unfractionated heparin on liver regeneration in rats after reduced-size liver transplantation.
【Methods】Try to set up stable Orthotopic Liver Transplantation model and serial reduced-size liver transplantation(60%,50%,30%) model in rats using two-cuff technique.To master the surgical skills of Liver transplantation in rats and reduce the complications.On this basis to select the same season,the same batch healthy male wistar rats 120(Beijing Academy of Military Medical Sciences),weighting 230-280g, were randomly divided into four groups(random number table method):A,B two sets were 42 rats each group,C,D were 18 in each group,the donors and receptors were randomly selected.50%reduced-size orthotopic liver transplantation was performed in A,B groups using two-cuff technique.The rats in A,B groups were given normal saline(4ml/kg) and unfractionated heparin(1000U/kg) respectively. 50%hepatectomy was performed in C group given saline after hepatectomy(4ml/kg). D group was sham-operated group,given normal saline(dose with the C group) postoperative.All of the drugs were administrated subcutaneous 20h postoperative. Rats in each group after 3 days,5 days,7 days were randomly sacrificed 6.Blood samples(5ml) were obtained from inferior vena cava of rat when Rats were sacrificed after anesthesia.Get serum after standing and centrifugalization,then put serum in the refrigerator(-80℃),and detect hepatocyte growth factor(HGF) by enzyme-linked immunosorbent assay(ELISA);check transplanted liver,Record rat weight,liver wet weight,the calculation of liver regeneration rate,make single-cell suspension from part of a liver by mechanical method;Detect Liver cell proliferation index by flow cytometer,part of a liver cryopreserved -80℃,and the remaining part of liver was fixed by formaldehyde,make conventional HE staining and immunohistochemical staining(liver cells proliferating nuclear antigen PCNA),Record PCNA expression. Results were analyzed using SPSS 13.0 statistical analysis software,observe compa rison of various indicators of group differences in liver regeneration(independent samples t test),and compare each group differences in the levels of HGF.
【Results】Complete one hundred cases of orthotopic liver transplantation in rat and a series of reduced-size liver transplantation(60%,50%,30%) before formal experiment,and set up a stable model of rat liver transplantation.Rats of group A and B were performed 50%reduced-size liver transplantation,the time for donor operation was(30±3)min and(31±4)min respectively,the recipient operation time was(54±7)min and(58±5)min,respectively,the anhepatic time was(15±2)min and(16±2)min,cold ischemia time was(56±5)min、(54±7)min respectively,the 48 hours survival rate was 95.2%(20/21)and 100%(21/21),no rats died of infection and bleeding.Two rats of group A were excluded due to biliary complications of postoperative,the incidence of biliary complications was 10%(2/20).Three cases of group B were excluded due to biliary complications of postoperative,the incidence of biliary complications was 14.3%(3/21).The rats of group C(18 cases ) were completed 50%hepatectomy,time for the operation was(14±2)min.There was no case of death and complications of group C and D.Liver regeneration rate of groups A,B,C on the third,fifth,seventh day after surgery was(33.5±6.6)%、(43.7±5.4)%、(45.6±6.3)%,(40.1±2.8)%、(71.9±17.8)%、(56.9±5.4)%,(86.6±11.9) %、(111±19.2)%、(98.4±4.6)%,respectively.There were significant differences(P = 0.015 <0.05,P= 0.002 <0.01,P= 0.023 <0.05) between A and B on the third,fifth, seventh day after surgery.There were significant differences(P=0.012<0.05,P= 0.011<0.05,P=0.031<0.05) between A and C on the third,fifth,seventh day after surgery.Serum HGF levels of groups A,B,C on the third,fifth,seventh day after surgery were(263.3±101.1)pg/ml、(420.7±56.8) pg/ml、(365.3±6.4)pg/ml, (312.9±10.1)pg/ml、(440.9±23.4)pg/ml、(384.4±32.2)pg/ml,(242.4±22.1) pg/ml、(409.9±23.0)pg/ml、(376.3±22.3)pg/ml.There were significant differences on serum HGF levels on the third,fifth,seventh day after surgery(P = 0.008 <0.01,P = 0.000 <0.01,P = 0.000 <0.01) between A and B.There were significant differences on serum HGF levels on the third,fifth,seventh day after surgery(P=0.016<0.05,P=0.038<0.05,P=0.001 <0.01) between A and C. Hepatocyte PI index of groups A,B,C on the third,fifth,seventh day after surgery were as follows:(30.4±4)%、(48.1±8)%、(38.4±2)%、(29.3±3)%、(43.5±8) %、(36.9±2)%,(30.3±3)%、(47.3±8)%、(31.6±3)%,there were significant differences on hepatocyte PI index on the third,fifth,seventh day after surgery(P = 0.001 <0.01,P = 0.002 <0.01,P = 0.02 <0.05)between A and B,and significant differences(P=0.025<0.05、P=0.018<0.05) on the third,fifth day between A and C,but have no difference on the seventh day between A and C(P=0.86>0.05).There were differences on PCNA expression between A and B on the third,fifth day,but no significant difference on PCNA expression on the seventh day.There were differences on PCNA expression between A and C on the fifth day,but no significant difference on PCNA expression on the third,seventh day.
【Conclusion】unfractionated heparin(1000IU/kg) given subcutaneous significantly promote hepatic regeneration after 50%reduced-size liver transplantation in rats;the liver regeneration of 50%reduced-size liver transplantation was delayed comparing to hepatectomy.In this experiment,it may be one of the mechanisms beneficial to liver regeneration that heparin elevates the plasma production of hepatocyte growth factor.
【方法】实验前期进行显微外科训练,尝试二袖套法建立大鼠全肝肝移植模型和系列减体积肝移植(60%、50%、30%)模型。在此基础上选取同季节健康雄性wistar大鼠120只(北京军事医学科学院实验动物中心),体重230-280g,随机分为四组(随机数表法):A、B两组每组42只,供受体随机选取,C、D两组每组18只。A、B两组行50%减体积肝移植术后分别给予生理盐水(4ml/kg)、肝素(1000IU/kg),C组行50%肝切除术后给予生理盐水(4ml/kg),D组为假手术组,术后给予生理盐水(剂量同C组),各组均为术后20h皮下给药。各组大鼠术后3天、5天、7天随机牺牲6只。牺牲鼠麻醉开腹后下腔静脉取血5ml,静置后离心取血清,酶联免疫吸附试验检测肝细胞生长因子(HGF);取移植肝脏,记录牺牲前大鼠重量、肝脏湿重,计算肝再生速率,取部分肝脏制作肝脏单细胞悬液检测肝细胞增殖指数,部分肝脏-80℃深低温保存,剩余肝脏甲醛固定,做常规HE染色和免疫组化染色(肝细胞增殖核抗原PCNA),记录PCNA表达情况。统计学分析比较A与B组、A与C组肝脏再生指标的差异(SPSS13.0,独立样本t检验),比较A与B组、A与C组HGF水平的差异。
【结果】前期完成一百余例大鼠全肝肝移植和系列减体积肝移植,建立了稳定的大鼠肝移植模型。A、B两组大鼠50%减体积肝移植术中供体切取时间分别为30±3min、31±4min,无肝期分别为15±2min、16±2min,冷缺血时间分别为56±5min、54±7min,受体手术时间分别为54±7min、58±5min。A、B两组术后48h存活率分别为95.2%(20/21)、100%(21/21),移植鼠无出血和感染死亡,A组两例因术后发生胆道并发症排除,胆道并发症发生率为10%(2/20),B组3例因术后发生胆道并发症排除,胆道并发症发生率14.3%(3/21)。C组完成大鼠50%肝切除18例,手术用时(14±2)min。C、D两组无死亡和并发症病例。A、B、C三组术后3天、5天、7天肝再生速率分别为(33.5±6.6)%、(43.7±5.4)%、(45.6±6.3)%,(40.1±2.8)%、(71.9±17.8)%、(56.9±5.4)%,(86.6±11.9)%、(111±19.2)%、(98.4±4.6)%。A、B两组术后3天、5天、7天肝再生速率比较均有统计学差异(P=0.015＜0.05、P=0.002＜0.01、P=0.023＜0.05)。A、C两组术后3天、5天、7天肝再生速率比较均有统计学差异(P=0.012＜0.05、P=0.011＜0.05、P=0.031＜0.05)。A、B、C组大鼠术后3天、5天、7天血清HGF水平分别为(263.3±101.1)pg/ml、(420.7±56.8)pg/ml、(365.3±6.4)pg/ml,(312.9±10.1)pg/ml、(440.9±23.4)pg/ml、(384.4±32.2)pg/ml,(242.4±22.1)pg/ml、(409.9±23.0)pg/ml、(376.3±22.3)pg/ml。A、B两组大鼠术后3天、5天、7天血清HGF水平比较有统计学差异(P=0.008＜0.01、P=0.000＜0.01、P=0.000＜0.01)。A、C两组术后3、5、7天血清HGF水平比较均有统计学差异(P=0.016＜0.05、P=0.038＜0.05、P=0.001＜0.01)。A、B、C组术后3天、5天、7天肝细胞增殖指数PI分别为:(30.4±4)%、(48.1±8)%、(38.4±2)%,(29.3±3)%、(43.5±8)%、(36.9±2)%,(30.3±3)%、(47.3±8)%、(31.6±3)%。A、B两组不同时间段PI指数水平比较均有统计学差异(P=0.001＜0.01、P=0.002＜0.01、P=0.02＜0.05)。A、C两组术后3、5天PI指数比较均有统计学差异(P=0.025＜0.05、P=0.018＜0.05),A、C两组术后7天PI指数比较无统计学差异(P=0.86＞0.05)。A、B两组术后3天、5天PCNA表达均有统计学差异,而术后7天PCNA表达无统计学差异。A、C两组术后3天、7天PCNA表达无统计学差异,术后5天PCNA表达有统计学差异。
【结论】皮下给予肝素(1000IU/kg)可促进大鼠50%减体积肝移植术后肝再生;大鼠减体积肝移植术后肝脏再生速率较肝切除术后肝再生延缓;本实验肝素促进血清HGF水平的升高可能是肝素促进肝再生的机制之一。
【Objective】Unfractionated heparin,low molecular weight heparin were regularly utilized to prevent or treat hepatic artery thrombosis during early postoperative in clinical living donor liver transplantation.However,there is few literatures on effects of heparin on hepatic regeneration after 50%partial liver transplantation in rats.This study was designed to investigate effects of unfractionated heparin on liver regeneration in rats after reduced-size liver transplantation.
【Methods】Try to set up stable Orthotopic Liver Transplantation model and serial reduced-size liver transplantation(60%,50%,30%) model in rats using two-cuff technique.To master the surgical skills of Liver transplantation in rats and reduce the complications.On this basis to select the same season,the same batch healthy male wistar rats 120(Beijing Academy of Military Medical Sciences),weighting 230-280g, were randomly divided into four groups(random number table method):A,B two sets were 42 rats each group,C,D were 18 in each group,the donors and receptors were randomly selected.50%reduced-size orthotopic liver transplantation was performed in A,B groups using two-cuff technique.The rats in A,B groups were given normal saline(4ml/kg) and unfractionated heparin(1000U/kg) respectively. 50%hepatectomy was performed in C group given saline after hepatectomy(4ml/kg). D group was sham-operated group,given normal saline(dose with the C group) postoperative.All of the drugs were administrated subcutaneous 20h postoperative. Rats in each group after 3 days,5 days,7 days were randomly sacrificed 6.Blood samples(5ml) were obtained from inferior vena cava of rat when Rats were sacrificed after anesthesia.Get serum after standing and centrifugalization,then put serum in the refrigerator(-80℃),and detect hepatocyte growth factor(HGF) by enzyme-linked immunosorbent assay(ELISA);check transplanted liver,Record rat weight,liver wet weight,the calculation of liver regeneration rate,make single-cell suspension from part of a liver by mechanical method;Detect Liver cell proliferation index by flow cytometer,part of a liver cryopreserved -80℃,and the remaining part of liver was fixed by formaldehyde,make conventional HE staining and immunohistochemical staining(liver cells proliferating nuclear antigen PCNA),Record PCNA expression. Results were analyzed using SPSS 13.0 statistical analysis software,observe compa rison of various indicators of group differences in liver regeneration(independent samples t test),and compare each group differences in the levels of HGF.
【Results】Complete one hundred cases of orthotopic liver transplantation in rat and a series of reduced-size liver transplantation(60%,50%,30%) before formal experiment,and set up a stable model of rat liver transplantation.Rats of group A and B were performed 50%reduced-size liver transplantation,the time for donor operation was(30±3)min and(31±4)min respectively,the recipient operation time was(54±7)min and(58±5)min,respectively,the anhepatic time was(15±2)min and(16±2)min,cold ischemia time was(56±5)min、(54±7)min respectively,the 48 hours survival rate was 95.2%(20/21)and 100%(21/21),no rats died of infection and bleeding.Two rats of group A were excluded due to biliary complications of postoperative,the incidence of biliary complications was 10%(2/20).Three cases of group B were excluded due to biliary complications of postoperative,the incidence of biliary complications was 14.3%(3/21).The rats of group C(18 cases ) were completed 50%hepatectomy,time for the operation was(14±2)min.There was no case of death and complications of group C and D.Liver regeneration rate of groups A,B,C on the third,fifth,seventh day after surgery was(33.5±6.6)%、(43.7±5.4)%、(45.6±6.3)%,(40.1±2.8)%、(71.9±17.8)%、(56.9±5.4)%,(86.6±11.9) %、(111±19.2)%、(98.4±4.6)%,respectively.There were significant differences(P = 0.015 <0.05,P= 0.002 <0.01,P= 0.023 <0.05) between A and B on the third,fifth, seventh day after surgery.There were significant differences(P=0.012<0.05,P= 0.011<0.05,P=0.031<0.05) between A and C on the third,fifth,seventh day after surgery.Serum HGF levels of groups A,B,C on the third,fifth,seventh day after surgery were(263.3±101.1)pg/ml、(420.7±56.8) pg/ml、(365.3±6.4)pg/ml, (312.9±10.1)pg/ml、(440.9±23.4)pg/ml、(384.4±32.2)pg/ml,(242.4±22.1) pg/ml、(409.9±23.0)pg/ml、(376.3±22.3)pg/ml.There were significant differences on serum HGF levels on the third,fifth,seventh day after surgery(P = 0.008 <0.01,P = 0.000 <0.01,P = 0.000 <0.01) between A and B.There were significant differences on serum HGF levels on the third,fifth,seventh day after surgery(P=0.016<0.05,P=0.038<0.05,P=0.001 <0.01) between A and C. Hepatocyte PI index of groups A,B,C on the third,fifth,seventh day after surgery were as follows:(30.4±4)%、(48.1±8)%、(38.4±2)%、(29.3±3)%、(43.5±8) %、(36.9±2)%,(30.3±3)%、(47.3±8)%、(31.6±3)%,there were significant differences on hepatocyte PI index on the third,fifth,seventh day after surgery(P = 0.001 <0.01,P = 0.002 <0.01,P = 0.02 <0.05)between A and B,and significant differences(P=0.025<0.05、P=0.018<0.05) on the third,fifth day between A and C,but have no difference on the seventh day between A and C(P=0.86>0.05).There were differences on PCNA expression between A and B on the third,fifth day,but no significant difference on PCNA expression on the seventh day.There were differences on PCNA expression between A and C on the fifth day,but no significant difference on PCNA expression on the third,seventh day.
【Conclusion】unfractionated heparin(1000IU/kg) given subcutaneous significantly promote hepatic regeneration after 50%reduced-size liver transplantation in rats;the liver regeneration of 50%reduced-size liver transplantation was delayed comparing to hepatectomy.In this experiment,it may be one of the mechanisms beneficial to liver regeneration that heparin elevates the plasma production of hepatocyte growth factor.
引文
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4.王学浩,杜竞辉,张峰等.活体供肝原位部分肝移植一例报告[J].中华器官移植杂志,1995,16(3):133-134.
5.Boyvat F,Aytekin C,Firat A,et al.Diagnostic and therapeutic management of hepatic artery thrombosis and stenosis after orthotopic and heterotopic liver transplantation[J].Transplant Proc,2003;35(7):2791-2795.
6.李岗山,叶启发.背驮式肝移植血管并发症的预防及处理[J]。中华临床医师杂志,2007,1(5):3-7。
7.王晨宇,刘连新,姜洪池。肝移植后肝动脉血栓形成的危险因素及防治措施[J]。中国普外基础与临床杂志,2007,14(5):516-519。
8.潘光栋,严律南.成人活体右半肝移植和小肝综合征[J].国际外科学杂志,2006,33(4):256-258.
9.马跃峰,李相成。成人间活体肝移植中小肝综合征的预防策略[J]。世界华人消化杂志,2005,13(18):2246-2250。
10.Kunio M,Toshikazu N.Heparin Functions as a Hepatotrophic Factor by Inducing Prod uction of Hepatocyte Growth Factor[J].Biochemical and biophysical research communications.1996,227:455-461.
11.SEIDE C,HANSEN H,WAAGE A,et al.Hepatocyte growth factor in serum after injectionof unfractionated and low molecular weight heparin in healthy individuals[J].British Journal of Haematology,1999,105,641-647.
12.Teizo N,Shigeki A,Masayuki I.Heparin accelerates liver regeneration following portal branch ligation in normal and cirrhotic rats with increased plasma hepatocyte growth factor levels[J].Journal of Hepatology,2002,(37):87-92.
13.Yuceturk H,Yagmurdur M C,et al.Roles of Heparin on TNF-α and IL-6 levels in liver regeneration after hepatic resection[J].Europ surgical.2007,39:216-221.
14.Lee S,Charters AC,Chandler JG,et al.A technique of orthotopic liver transplanta-tion in the rat[J].Transplantation,1973,16(6):664-666.
15.Kamada N,Calne RY.A surgical experience with five hundred thirty liver transplants in the rats[J].Surgery,1983,93(1):64-69.
16.Miyata M,Fischer JH,Fuhsm M,et al.A simple method for orthotopic liver transplantation in the rat[J].Yransplantation.1980,30(5):335-338.
17.彭勇,龚建平,严律南等。大鼠原位肝移植模型制作过程中麻醉方法的选择[J]。中国普通外科杂志,2003,12(9):673-676.
18.刘金彪,郑树国,陈丽君.提高大鼠原位肝移植手术质量的技巧和注意事项[J].中华器官移植杂志,2005,26(8):500-502.
19.李军,窦科峰,张洪涛等.供肝快速切取法大鼠原位肝移植模型的建立[J].中国普通外科杂志,2006,15(7):501-503.
20.马跃峰,李相成,游伟.两袖套法大鼠原位肝移植模型的技术改进[J].中国现代医学杂志,2006,16(2):229-231.
21.宋康,樊嘉,汤钊酞等。大鼠部分肝移植模型的建立[J].复旦学报(医学版),2007,34(2):307-309.
22.叶晟,韩本立,董家鸿。大鼠系列减体积原位肝移植的实验研究[J].肝胆外科杂志,2003,11(3):220-222.
23.梁廷波,郑树森,范上达等.两种不同方法的大鼠小体积肝移植模型建立的比较[J].中华肝胆外科杂志.2004,10(5):320-322.
24.汪泳,宇汝胜,钱海鑫。大鼠原位肝移植模型手术并发症的分析[J].肝胆外科杂志,2008,16(5):355-357.
25.李涛,孙星,彭志海.大鼠肝移植术后并发症的预防[J].肝胆胰外科杂志.2007,19(4):215-217.
26.邰升,苏志雷,包少瑜等.大鼠原位肝移植模型不同术式改进的研究[J].中华肝胆外科杂志,2006,12(5):333-335.
27.Dowing LJ,Strieter RM,Kadell AM,et al.Low-dose low-molecular weight heparin is anti-inflammatory during venous thrombosis[J].J Vase Surg,1998,28(5):848-854.
28.Manduteanu I,Voinea M,Capraru M,et al.A novel attribute of enoxapa rin:inhibition o fm onocytea dhesiont oe ndothelialc else by a mechanism in volvingc el adhesion molecules[J].Pharmacology,2002,65(1):32-37.
29.Yamabe H,Osawa H,Inuma H,et al.Some structural determinants of the antiproliferative effect of heparin-like molecules on human airway smooth muscle[J].Thromb Haemost,1996,76(2):215-219.
30.Moreno E,Meneu JC,delaCalle A,et al.Modulation of Hepatocyte Growth Factor Plasma Levels in Relation to the Dose of Exogenous Heparin Administered:An Experimental Study in Rats[J].Transplantation Proceedings,2005,37:3943-3947.
31.Akira Matsumori,Koh Ono,Shigetake Sasayama,et al.Immediate Increase in Cir culating Hepatocyte Growth Factor/Scatter Factor by Heparin[J].J Mol Cell Cardiol,1998,30:2145-2149.
32.李宁,赵晓飞,周天琼等.促肝细胞生长素肠溶胶囊与其注射剂对大鼠肝切除后肝再生影响的对照实验研究[J].中国现代应用药学,2008,25(6):495-498.
33.Nimer Assy,Osamah Hussein,Gadi Spira et al.The Beneficial Effect of Aspirin and Enoxaparin on Fibrosis Progression and Regenerative Activity in a Rat Model of Cirrhosis[J].Dig Dis Sci,2007,52:1187-1193.
34.CARINA S,HENRIK H H,ANDERS WAAGE.Hepatocyte growth factor in serum after injection of unfractionated and low molecular weight heparin in healthy individuals[J].British Journal o f Haematology,1999,105,641-647。
35.Okada,K;Ueshima,S;Kawao,N,et al.Binding of plasminogen to hepatocytes isolated from injured mice liver and nonparenchymal cell-dependent proliferation of hepatocytes[J].Blood coagulation & fibrinolysis,2008,19(6):503-511.
1.Eleftheriadis,Theodoros,Ioannis,etal.About the effect of low-molecular-weight heparin on platelet function in haemodialysis patients[J].Nephrol Dial Transplant,2009,24(3):1063-1064.
2.Broughton-Head VJ,Shur J,Shute JK,et al.Unfractionated heparin reduces the elasticity of sputum from patients with cystic fibrosis.American journal of physiology-lung cellular and molecular physiology[J].2007,293(5):1240-1249.
3.Hach-Wunderle V.Diagnosis and treatment of venous thrombosis[J].Hamostaseologie,2008,28(5):376-386.
4.Davenport A.Low-molecular-weight heparin for routine hemodialysis[J]. Hemodial Int.2008,12(2):S34-37.
5.Michalopoulos G K,DeFrances M C.Liver regeneration[J].Science,1997,276:60-66.
6.Rebecca Taub.liver regeneration:from myth to mechanism[J].Nature review molecular cell biology,2004,5:836-847.
7.KHAN A Z,MUDAN S S.Liver regeneration:mechanisms,mysteries,and more[J].ANZ J.Surg.2007,77:9-14.
8.FuastoN.Liver regeneration[J].JHepatol,2000:32:19-31.
9.Widman JJ,Fahimi HD.Proliferation of mononuclear phagocytes(Kupffer cells)and endothelial cells in regeneration rat liver[J].Am.J.Pathol.1975;80:349-66.
10.Tajima F,Tsuchitya H,Shiota G.Contribution of HGF to liver regeneration by strong mobilization of bone marrow stem cells[J].Blood.2006,108(11):409-410.
11.Little EB,Hornyak SC,Radi A,Gehlsen KR,Haaparanta T.Histamine treatment accelerates liver regeneration following partial hepatectomy by up-regulating TGF-alpha despite pretreatment with lipopolysaccharide[J].HEPATOLOGY,2003,38(4):545-547.
12.Cressman DE,Greenbaumm LE,DeAngelis RA et al.Liver failure and defective hepatocyte regeneration in interleukin-6 deficient mice[J].Science 1996;274:1379-1383.
13.Court F G,Wemyss-Holden S A,Maddern G J,et al.The mystery of liver regeneration[J].British Journal of Surgery,2002,89:1089-1095.
14.Zhong Z,Tsukada S,Lemasters JJ,et al.TGF-beta suppresses regeneration of small-for-size liver grafts by activating Smad and P21(CIP1) pathways[J].HEPATOLOGY,2006,44(4):218-222.
15.FuastoN.Liverergeneration[J].Jhepatol,2000:32:19-31.
16.顾文杨。肝动脉重建对大鼠部分肝移植后移植肝的影响[D]。南京:南京医科大学,2005:40-45。
17.杨靖.半肝切除术后血清白细胞介素2、白细胞介素6、白细胞介素8、肿瘤坏死因子及胰岛素样生长因子变化[J].中国组织工程研究与临床康复. 2008,12(18):3483-3486.
18.Haga S,Ogawa W,Ozaki M,et al.Compensatory recovery of liver mass by Akt-mediated hepatocellular hypertrophy in liver-specific STAT3-deficient mice[J].Journal ofhepatology,2005,43(5):799-807.
19.汪谦,李湘竑,陈俊任。PPARα和Bcl-XL在体外肝手术后肝组织中的表达及意义[J].中华肝胆外科杂志,2005,11(12):832-836.
20.Kiba,TSaitoS,ShiraishiM,etal.Fas(APO-1/CD95)mRNA Is down-reuglated in liver regeneration after hepatectomy in rats[J].J Gastroenterol,2000;35:34-38.
21.TairaK,HiroyasuS,ShiarishiM,et al.Epxression of bcl-2 family reudces apoptotic hepatocytes after exeessive hepatectomy[J].Eur SurgRes,2001,33:334-341.
22.Desbarats J,Newell M K.Fas engagement accelerates liver regeneration after partial hepatoetomy[J].Nat Med,2000,6:920-923.
23.Boyvat F,Aytekin C,Firat A,et al.Diagnostic and therapeuticmanagement of hepatic artery t hrombosis and stenosis after orthotopic and heterotopic liver transplantation[J].Transplant Proc,2003;35(7):2791-2795.
24.李岗山,叶启发.背驮式肝移植血管并发症的预防及处理[J].中华临床医师杂志,2007,1(5):3-7。
25.Kunio M,Toshikazu N.Heparin Functions as a Hepatotrophic Factor by Inducing Production of Hepatocyte Growth Factor[J].Biochemical and biophysical research communications.1996,227:455-461.
26.Akira Matsumori,Koh Ono,Shigetake Sasayama,et al.Immediate Increase in Circulating Hepatocyte Growth Factor/Scatter Factor by Heparin[J].J Mol Cell Cardiol,1998,30:2145-2149.
27.Teizo N,Shigeki A,Masayuki I.Heparin accelerates liver regeneration following portal branch ligation in normal and cirrhotic rats with increased plasma hepatocyte growth factor levels[J].Journal of Hepatology,2002,(37):87-92.
28.CARINA S,HENRIK H H,ANDERS WAAGE.Hepatocyte growth factor in serum after injection of unfractionated and low molecular weight heparin in healthy individuals[J].British Journal o f Haematology,1999,105,641-647.
29.Okada,K;Ueshima,S;Kawao,N,et al.Binding of plasminogen to hepatocytes isolated from injured mice liver and nonparenchymal cell-dependent proliferation of hepatocytes [J].Blood coagulation & fibrinolysis,2008,19(6):503-511.
30.Nimer Assy,Osamah Hussein,Gadi Spira et al.The Beneficial Effect of Aspirin and Enoxaparin on Fibrosis Progression and Regenerative Activity in a Rat[J].Model of Cirrhosis.Dig Dis Sci,2007,52:1187-1193.
31.Yuceturk H,Yagmurdur M C,et al.Roles of Heparin on TNF-aand IL-6 levels in liver regeneration after hepatic resection[J].Europ surgical.2007,39:216-221.
2.Strong RW,Lynch SV.Ethical issues in living related donor liver transplantation[J].Transplant Proc,1996,28:2366-2369.
3.Lo C,Fan S T,et al.Adult-to-Adult Living Donor Liver Transplantation Using Extended Right Lobe Grafts[J].Annals of surgery,1997,226(3):261-270.
4.王学浩,杜竞辉,张峰等.活体供肝原位部分肝移植一例报告[J].中华器官移植杂志,1995,16(3):133-134.
5.Boyvat F,Aytekin C,Firat A,et al.Diagnostic and therapeutic management of hepatic artery thrombosis and stenosis after orthotopic and heterotopic liver transplantation[J].Transplant Proc,2003;35(7):2791-2795.
6.李岗山,叶启发.背驮式肝移植血管并发症的预防及处理[J]。中华临床医师杂志,2007,1(5):3-7。
7.王晨宇,刘连新,姜洪池。肝移植后肝动脉血栓形成的危险因素及防治措施[J]。中国普外基础与临床杂志,2007,14(5):516-519。
8.潘光栋,严律南.成人活体右半肝移植和小肝综合征[J].国际外科学杂志,2006,33(4):256-258.
9.马跃峰,李相成。成人间活体肝移植中小肝综合征的预防策略[J]。世界华人消化杂志,2005,13(18):2246-2250。
10.Kunio M,Toshikazu N.Heparin Functions as a Hepatotrophic Factor by Inducing Prod uction of Hepatocyte Growth Factor[J].Biochemical and biophysical research communications.1996,227:455-461.
11.SEIDE C,HANSEN H,WAAGE A,et al.Hepatocyte growth factor in serum after injectionof unfractionated and low molecular weight heparin in healthy individuals[J].British Journal of Haematology,1999,105,641-647.
12.Teizo N,Shigeki A,Masayuki I.Heparin accelerates liver regeneration following portal branch ligation in normal and cirrhotic rats with increased plasma hepatocyte growth factor levels[J].Journal of Hepatology,2002,(37):87-92.
13.Yuceturk H,Yagmurdur M C,et al.Roles of Heparin on TNF-α and IL-6 levels in liver regeneration after hepatic resection[J].Europ surgical.2007,39:216-221.
14.Lee S,Charters AC,Chandler JG,et al.A technique of orthotopic liver transplanta-tion in the rat[J].Transplantation,1973,16(6):664-666.
15.Kamada N,Calne RY.A surgical experience with five hundred thirty liver transplants in the rats[J].Surgery,1983,93(1):64-69.
16.Miyata M,Fischer JH,Fuhsm M,et al.A simple method for orthotopic liver transplantation in the rat[J].Yransplantation.1980,30(5):335-338.
17.彭勇,龚建平,严律南等。大鼠原位肝移植模型制作过程中麻醉方法的选择[J]。中国普通外科杂志,2003,12(9):673-676.
18.刘金彪,郑树国,陈丽君.提高大鼠原位肝移植手术质量的技巧和注意事项[J].中华器官移植杂志,2005,26(8):500-502.
19.李军,窦科峰,张洪涛等.供肝快速切取法大鼠原位肝移植模型的建立[J].中国普通外科杂志,2006,15(7):501-503.
20.马跃峰,李相成,游伟.两袖套法大鼠原位肝移植模型的技术改进[J].中国现代医学杂志,2006,16(2):229-231.
21.宋康,樊嘉,汤钊酞等。大鼠部分肝移植模型的建立[J].复旦学报(医学版),2007,34(2):307-309.
22.叶晟,韩本立,董家鸿。大鼠系列减体积原位肝移植的实验研究[J].肝胆外科杂志,2003,11(3):220-222.
23.梁廷波,郑树森,范上达等.两种不同方法的大鼠小体积肝移植模型建立的比较[J].中华肝胆外科杂志.2004,10(5):320-322.
24.汪泳,宇汝胜,钱海鑫。大鼠原位肝移植模型手术并发症的分析[J].肝胆外科杂志,2008,16(5):355-357.
25.李涛,孙星,彭志海.大鼠肝移植术后并发症的预防[J].肝胆胰外科杂志.2007,19(4):215-217.
26.邰升,苏志雷,包少瑜等.大鼠原位肝移植模型不同术式改进的研究[J].中华肝胆外科杂志,2006,12(5):333-335.
27.Dowing LJ,Strieter RM,Kadell AM,et al.Low-dose low-molecular weight heparin is anti-inflammatory during venous thrombosis[J].J Vase Surg,1998,28(5):848-854.
28.Manduteanu I,Voinea M,Capraru M,et al.A novel attribute of enoxapa rin:inhibition o fm onocytea dhesiont oe ndothelialc else by a mechanism in volvingc el adhesion molecules[J].Pharmacology,2002,65(1):32-37.
29.Yamabe H,Osawa H,Inuma H,et al.Some structural determinants of the antiproliferative effect of heparin-like molecules on human airway smooth muscle[J].Thromb Haemost,1996,76(2):215-219.
30.Moreno E,Meneu JC,delaCalle A,et al.Modulation of Hepatocyte Growth Factor Plasma Levels in Relation to the Dose of Exogenous Heparin Administered:An Experimental Study in Rats[J].Transplantation Proceedings,2005,37:3943-3947.
31.Akira Matsumori,Koh Ono,Shigetake Sasayama,et al.Immediate Increase in Cir culating Hepatocyte Growth Factor/Scatter Factor by Heparin[J].J Mol Cell Cardiol,1998,30:2145-2149.
32.李宁,赵晓飞,周天琼等.促肝细胞生长素肠溶胶囊与其注射剂对大鼠肝切除后肝再生影响的对照实验研究[J].中国现代应用药学,2008,25(6):495-498.
33.Nimer Assy,Osamah Hussein,Gadi Spira et al.The Beneficial Effect of Aspirin and Enoxaparin on Fibrosis Progression and Regenerative Activity in a Rat Model of Cirrhosis[J].Dig Dis Sci,2007,52:1187-1193.
34.CARINA S,HENRIK H H,ANDERS WAAGE.Hepatocyte growth factor in serum after injection of unfractionated and low molecular weight heparin in healthy individuals[J].British Journal o f Haematology,1999,105,641-647。
35.Okada,K;Ueshima,S;Kawao,N,et al.Binding of plasminogen to hepatocytes isolated from injured mice liver and nonparenchymal cell-dependent proliferation of hepatocytes[J].Blood coagulation & fibrinolysis,2008,19(6):503-511.
1.Eleftheriadis,Theodoros,Ioannis,etal.About the effect of low-molecular-weight heparin on platelet function in haemodialysis patients[J].Nephrol Dial Transplant,2009,24(3):1063-1064.
2.Broughton-Head VJ,Shur J,Shute JK,et al.Unfractionated heparin reduces the elasticity of sputum from patients with cystic fibrosis.American journal of physiology-lung cellular and molecular physiology[J].2007,293(5):1240-1249.
3.Hach-Wunderle V.Diagnosis and treatment of venous thrombosis[J].Hamostaseologie,2008,28(5):376-386.
4.Davenport A.Low-molecular-weight heparin for routine hemodialysis[J]. Hemodial Int.2008,12(2):S34-37.
5.Michalopoulos G K,DeFrances M C.Liver regeneration[J].Science,1997,276:60-66.
6.Rebecca Taub.liver regeneration:from myth to mechanism[J].Nature review molecular cell biology,2004,5:836-847.
7.KHAN A Z,MUDAN S S.Liver regeneration:mechanisms,mysteries,and more[J].ANZ J.Surg.2007,77:9-14.
8.FuastoN.Liver regeneration[J].JHepatol,2000:32:19-31.
9.Widman JJ,Fahimi HD.Proliferation of mononuclear phagocytes(Kupffer cells)and endothelial cells in regeneration rat liver[J].Am.J.Pathol.1975;80:349-66.
10.Tajima F,Tsuchitya H,Shiota G.Contribution of HGF to liver regeneration by strong mobilization of bone marrow stem cells[J].Blood.2006,108(11):409-410.
11.Little EB,Hornyak SC,Radi A,Gehlsen KR,Haaparanta T.Histamine treatment accelerates liver regeneration following partial hepatectomy by up-regulating TGF-alpha despite pretreatment with lipopolysaccharide[J].HEPATOLOGY,2003,38(4):545-547.
12.Cressman DE,Greenbaumm LE,DeAngelis RA et al.Liver failure and defective hepatocyte regeneration in interleukin-6 deficient mice[J].Science 1996;274:1379-1383.
13.Court F G,Wemyss-Holden S A,Maddern G J,et al.The mystery of liver regeneration[J].British Journal of Surgery,2002,89:1089-1095.
14.Zhong Z,Tsukada S,Lemasters JJ,et al.TGF-beta suppresses regeneration of small-for-size liver grafts by activating Smad and P21(CIP1) pathways[J].HEPATOLOGY,2006,44(4):218-222.
15.FuastoN.Liverergeneration[J].Jhepatol,2000:32:19-31.
16.顾文杨。肝动脉重建对大鼠部分肝移植后移植肝的影响[D]。南京:南京医科大学,2005:40-45。
17.杨靖.半肝切除术后血清白细胞介素2、白细胞介素6、白细胞介素8、肿瘤坏死因子及胰岛素样生长因子变化[J].中国组织工程研究与临床康复. 2008,12(18):3483-3486.
18.Haga S,Ogawa W,Ozaki M,et al.Compensatory recovery of liver mass by Akt-mediated hepatocellular hypertrophy in liver-specific STAT3-deficient mice[J].Journal ofhepatology,2005,43(5):799-807.
19.汪谦,李湘竑,陈俊任。PPARα和Bcl-XL在体外肝手术后肝组织中的表达及意义[J].中华肝胆外科杂志,2005,11(12):832-836.
20.Kiba,TSaitoS,ShiraishiM,etal.Fas(APO-1/CD95)mRNA Is down-reuglated in liver regeneration after hepatectomy in rats[J].J Gastroenterol,2000;35:34-38.
21.TairaK,HiroyasuS,ShiarishiM,et al.Epxression of bcl-2 family reudces apoptotic hepatocytes after exeessive hepatectomy[J].Eur SurgRes,2001,33:334-341.
22.Desbarats J,Newell M K.Fas engagement accelerates liver regeneration after partial hepatoetomy[J].Nat Med,2000,6:920-923.
23.Boyvat F,Aytekin C,Firat A,et al.Diagnostic and therapeuticmanagement of hepatic artery t hrombosis and stenosis after orthotopic and heterotopic liver transplantation[J].Transplant Proc,2003;35(7):2791-2795.
24.李岗山,叶启发.背驮式肝移植血管并发症的预防及处理[J].中华临床医师杂志,2007,1(5):3-7。
25.Kunio M,Toshikazu N.Heparin Functions as a Hepatotrophic Factor by Inducing Production of Hepatocyte Growth Factor[J].Biochemical and biophysical research communications.1996,227:455-461.
26.Akira Matsumori,Koh Ono,Shigetake Sasayama,et al.Immediate Increase in Circulating Hepatocyte Growth Factor/Scatter Factor by Heparin[J].J Mol Cell Cardiol,1998,30:2145-2149.
27.Teizo N,Shigeki A,Masayuki I.Heparin accelerates liver regeneration following portal branch ligation in normal and cirrhotic rats with increased plasma hepatocyte growth factor levels[J].Journal of Hepatology,2002,(37):87-92.
28.CARINA S,HENRIK H H,ANDERS WAAGE.Hepatocyte growth factor in serum after injection of unfractionated and low molecular weight heparin in healthy individuals[J].British Journal o f Haematology,1999,105,641-647.
29.Okada,K;Ueshima,S;Kawao,N,et al.Binding of plasminogen to hepatocytes isolated from injured mice liver and nonparenchymal cell-dependent proliferation of hepatocytes [J].Blood coagulation & fibrinolysis,2008,19(6):503-511.
30.Nimer Assy,Osamah Hussein,Gadi Spira et al.The Beneficial Effect of Aspirin and Enoxaparin on Fibrosis Progression and Regenerative Activity in a Rat[J].Model of Cirrhosis.Dig Dis Sci,2007,52:1187-1193.
31.Yuceturk H,Yagmurdur M C,et al.Roles of Heparin on TNF-aand IL-6 levels in liver regeneration after hepatic resection[J].Europ surgical.2007,39:216-221.