单人直视下改良建立大鼠原位肝移植模型的体会
|
摘要
目的:建立稳定大鼠原位肝移植模型,缩短术中无肝期时间,提高手术成功率及受体存活率。方法:在Kamada"二袖套法"的基础上改进,单人直视下建立大鼠原位肝移植模型,行60例SD大鼠原位肝移植手术。本研究简化供受体麻醉方式,供肝采用经门静脉(必要时配合腹主动脉补救方式)进行冷灌注,缩短修肝时间,提前预置牵引线,固定进针位置,改进植入肝脏肝上下腔静脉吻合、肝下下腔静脉及门静脉套管。观察并记录各组大鼠供体手术、修肝套管、无肝期、受体手术及肝移植手术总时间。术后检测1,7,30天受体大鼠肝功能(血清丙氨酸转氨酶(ALT),天冬氨酸转氨酶(AST)及总胆红素(TB))并分析生存情况。结果:无肝期结束后,供体肝脏灌注良好,受体麻醉移除后较快苏醒。供体手术、修肝套管、无肝期、受体手术及肝移植手术总时间分别为(32.5±1.58)、(7.3±1.43)、(15.6±2.62)、(53.2±3.74)、(108.5±2.34)min。大鼠术后24 h(手术成功率)为95%,1周生存率分别为90%,1月生存率分别为86.7%。大鼠术后短时间内肝功能水平增高,24 h时ALT(228.5±54.5 IU/L),AST(439.3±86.3 IU/L),TB(6.2±0.7μM),1周后逐渐恢复正常。结论:改良后的方法可以简易麻醉流程,缩短无肝期,提高手术成功率及受体的生存率。
Objective: To establish a stable rat orthotopic liver transplantation model, reduce the time of anhepatic phase, improve the operation success and receptor survival rate. Methods: We make improvement of the Kamada's "two cuff method", the rat orthotopic liver transplantation model was established under the direct vision of single operator, and 60 cases were performed in the SD rats. We simplified the method of anesthesia for the donor and recipient, the donor liver was subjected to cold perfusion via the portal vein(if necessary with the abdominal aorta), reduced the donor liver repair time, an improved anastomosis of the suprahepatic and inferior vena cava was performed, the inferior hepatic vena cava and portal vein were cannulated by improving technique. Observe and record the time of donor surgery, mend liver and cannula, anhepatic phase, recipient surgery and the total time of liver transplantation. Liver function of recipient rats(including serum alanine aminotransferase(ALT), aspartate aminotransferase(AST) and total bilirubin(TB)) at 1, 7, and 30 days were detected and analyzed, their survival was also analyzed. Results: When the anhepatic period was over, the donor liver perfusion was perfused well and the receptor recovered quickly after the anesthesia removed. The time of donor operation, liver repair cannula,anhepatic phase, recipient operation and total time of liver transplantation were(32.5 ±1.58),(7.3±1.43),(15.6±2.62),(53.2±3.74),and(108.5±2.34) min respectively. After operation, the success rate of operation was 95 %, the one-week survival rate was 90 %, and the one-month survival rate was 86.7 %. The level of liver function increased in a short time after operation, at 24 h after operation the ALT/AST/TB in serum were detected and the values were(228.5±54.5 IU/L)/(439.3±86.3 IU/L)/(6.2±0.7 μM), then the liver function gradually returned to normal after one week. Conclusions: The modified method could simplify the anesthesia progress, shorten the period of anhepatic, improve the success rate of the operation and the survival rate of the receptor.
Objective: To establish a stable rat orthotopic liver transplantation model, reduce the time of anhepatic phase, improve the operation success and receptor survival rate. Methods: We make improvement of the Kamada's "two cuff method", the rat orthotopic liver transplantation model was established under the direct vision of single operator, and 60 cases were performed in the SD rats. We simplified the method of anesthesia for the donor and recipient, the donor liver was subjected to cold perfusion via the portal vein(if necessary with the abdominal aorta), reduced the donor liver repair time, an improved anastomosis of the suprahepatic and inferior vena cava was performed, the inferior hepatic vena cava and portal vein were cannulated by improving technique. Observe and record the time of donor surgery, mend liver and cannula, anhepatic phase, recipient surgery and the total time of liver transplantation. Liver function of recipient rats(including serum alanine aminotransferase(ALT), aspartate aminotransferase(AST) and total bilirubin(TB)) at 1, 7, and 30 days were detected and analyzed, their survival was also analyzed. Results: When the anhepatic period was over, the donor liver perfusion was perfused well and the receptor recovered quickly after the anesthesia removed. The time of donor operation, liver repair cannula,anhepatic phase, recipient operation and total time of liver transplantation were(32.5 ±1.58),(7.3±1.43),(15.6±2.62),(53.2±3.74),and(108.5±2.34) min respectively. After operation, the success rate of operation was 95 %, the one-week survival rate was 90 %, and the one-month survival rate was 86.7 %. The level of liver function increased in a short time after operation, at 24 h after operation the ALT/AST/TB in serum were detected and the values were(228.5±54.5 IU/L)/(439.3±86.3 IU/L)/(6.2±0.7 μM), then the liver function gradually returned to normal after one week. Conclusions: The modified method could simplify the anesthesia progress, shorten the period of anhepatic, improve the success rate of the operation and the survival rate of the receptor.
引文
[1]Welch CS.Liver graft[J].Maroc medical,1955,34(359):514-515
[2]Starzl TE,Marchioro TL,Vonkaulla KN,et al.Homotransplantation of the liver in humans[J].Surgery,gynecology&obstetrics,1963,117:659-676
[3]Lee S,Charters AC,Chandler JG,et al.A technique for orthotopic liver transplantation in the rat[J].Transplantation,1973,16(6):664-669
[4]Kamada N,Calne RY.Orthotopic liver transplantation in the rat.Technique using cuff for portal vein anastomosis and biliary drainage[J].Transplantation,1979,28(1):47-50
[5]陈忠华,夏穗生.改进的大鼠原位肝移植术[J].武汉医学院学报,1984,05(04):244Chen Zhong-hua,Xia Shui-sheng.Modification of rat orthotopic liver transplatation model[J].Journal of Wu han Yi Xue Yuan,1984,05(04):244
[6]徐峰,杨甲梅,吴孟超,等.改良Kamada法大鼠原位肝移植模型的建立[J].中华肝胆外科杂志,2001,7(12):758-759Xu Feng,Yang Jia-mei,Wu Meng-chao,et al.Modification of Kamada technique in the rat model of orthotopic liver transplantation[J].Chinese Journal of Hepatobiliary Surgery,2001,7(12):758-759
[7]Nagai K,Yagi S,Uemoto S,et al.Surgical procedures for a rat model of partial orthotopic liver transplantation with hepatic arterial reconstruction[J].Journal of Visualized Experiments,2013,37(73):e4376
[8]Kakizaki Y,Miyagi S,Shimizu K,et al.The effects of short-term subnormothermic perfusion after cold preservation on liver grafts from donors after cardiac death:an ex vivo rat model[J].Transplantation,2018,15
[9]Chen X,Zheng J,Cai J,et al.The cytoskeleton proteinβ-actin may mediate T cell apoptosis during acute rejection reaction after liver transplantation in a rat model[J].Am J Transl Res,2017,9(11):4888-4901
[10]Shinya O,Tadahiro U,Zhao Xiang-dong,et al.Liver graft preservation using perfluorocarbon improves the outcomes of simulated donation after cardiac death liver transplantation in rats[J].Liver Transpl,2017,23(6):804-812
[11]马毅,何晓顺,陈规划.大鼠原位肝移植模型的手术技巧及并发症的预防[J].中华显微外科杂志,2003,26(1):45-47Ma Yi,He Xiao-shun,Chen Gui-hua.Surgical technique of the model of orthotopic liver transplantation and prevention of operational complication in rat[J].Chinese Journal of Microsurgery,2003,26(1):45-47
[12]Wang R,Shen Z,Yang L,et al.Protective effects of heme oxygenase-1-transduced bone marrow-derived mesenchymal stem cells on reduced size liver transplantation:role of autophagy regulated by the ERK/m TOR signaling pathway[J].Int J Mol Med,2017,40(5):1537-1548
[13]刘立,方成,王加谋.丙泊芬、戊巴比妥钠和乌拉坦对肾移植大鼠麻醉效果的比较[J].咸宁学院学报(医学版),2012,26(4):277-278Liu Li,Fang Cheng,Wang Jia-mou.Comparison on anaesthetic effect between propofol,sodium pentobarbital and urethane in rats with kidney transplantation[J].Journal of Xianning Univarsity(medical Sciences),2012,26(4):277-278
[14]陈辉,姜骊,王雁,等.三种不同麻醉药对SD大鼠肝移植手术麻醉效果的比较和应用[J].实验动物科学,2009,26(4):57-59Chen Hui,Jiang Li,Wang Yan,et al.Effect of three anesthetic drugs in SD rat liver transplantation surgery[J].Laboratory Animal Science,2009,26(4):57-59
[15]Cao D,Liu Y,Li J,et al.Isoflurane:an ideal anesthetic for rodent orthotopic liver transplantation surgery?[J].Transplant Proc,2016,48(8):2815-2820
[16]Luo C,Yuan D,Li X,et al.Propofol attenuated acute kidney injury after orthotopic liver transplantation via inhibiting gap junction composed of connexin 32[J].Anesthesiology,2015,122(1):72-86
[17]Schemmer P,Schoonhoven R,Swenberg JA,et al.Gentle in situ liver manipulation during organ harvest decreases survival after rat liver transplantation:role of Kupffer cells[J].Transplantation,1998,65(8):1015-1020
[18]Zeng XP,Li XJ,Zhang QY,et al.Tert-butylhydroquinone protects liver against ischemia reperfusion injury in rats through nrf2-activating anti-oxidative activity[J].Transplant Proc,2017,49(2):366-372
[19]Ishii E,Shimizu A,Takahashi M,et al.Surgical technique of orthotopic liver transplantation in rats:the Kamada technique and a new splint technique for hepatic artery reconstruction[J].Journal of Nippon Medical School,2013,80(1):4-15
[20]Zhi X,Xue F,Chen W,et al.OSI-027 modulates acute graft-versus-host disease after liver transplantation in a rat model[J].Liver Transpl,2017,23(9):1186-1198
[21]葛勇胜,许戈良,李建生,等.供肝的不同灌注方法在大鼠肝移植中的应用[J].实用医学杂志,2006,22(22):2585-2587Ge Yong-sheng,Xu Ge-liang,Li Jian-sheng,et al.Different hepatic perfusion procedures for donor liver transplantation[J].The Journal of PracticalMedicine,2006,22(22):2585-2587
[22]许赤,杨扬,易述红,等.稳定大鼠肝移植模型的规范及移植肝灌注方式比较[J].南方医科大学学报,2006,26(11):1556-1558Xu Chi,Yang Yang,Yi Shu-hong,et al.Standardization of rat stable orthotopic liver transplantation model and comparison of the effect of two liver graft perfusion methods[J].Journal of Southern Medical University,2006,26(11):1556-1558
[23]陈强星,李坤,孔伟浩,等.单人直视下改良建立大鼠原位肝移植模型[J].中华肝脏外科手术学电子杂志,2017,6(2):127-133Chen Qiang-xing,Li Kun,Kong Wei-hao,et al.Modi覱ed approach for establishment of rat models with orthotopic liver transplantation under direct vision of single operator[J].Chin J Hepat Surg(Electronic Edition),2017,6(2):127-133
[24]Czigany Z,Iwasaki J,Yagi S,et al.Improving research practice in rat orthotopic and partial orthotopic liver transplantation:a review,recommendation,and publication guide[J].European surgical research Europaische chirurgische Forschung Recherches chirurgicales europeennes,2015,55(1-2):119-138
[25]Yu E,Ueta H,Kimura H,et al.Graft-versus-host disease following liver transplantation:development of a high-incidence rat model and a selective prevention method[J].Am J Transplant,2017,17(4):979-991
[26]Ma B,Yang JY,Song WJ,et al.Combining Exosomes Derived from Immature DCs with donor antigen-specific treg cells induces tolerance in a rat liver allograft model[J].Sci Rep,2016,6:32971
[27]Tang J,Yang R,Lv L,et al.Transforming growth factor-β-expressing mesenchymal stem cells induce local tolerance in a rat liver transplantation model of acute rejection[J].Stem Cells,2016,34(11):2681-2692
[28]白建华,陈刚,朱新锋,等.原位肝移植模型的建立技巧与改进[J].中国组织工程研究,2015,19(40):6526-6530Bai Jian-hua,Chen Gang,Zhu Xin-feng,et al.Techniques and improvements of establishing orthotopic liver transplantation rat models[J].Chinese Journal of Tissue Engineering Research,2015,19(40):6526-6530
[29]Yoshimura N,Matsui S,Hamashima T,et al.The effects of perioperative portal venous inoculation with donor lymphocytes on renal allograft survival in the rat.I.specific prolongation of donor grafts and suppressor factor in the serum[J].Transplantation,1990,49(1):167-71
[30]许永刚,翁明哲,徐军明,等.单人建立大鼠原位肝移植模型的手术体会[J].现代生物医学进展,2012,12(16):3026-3028Xu Yong-gang,Weng Ming-zhe,Xu Jun-ming,et al.Technique of orthotopic liver transplantation in rats operated by one person without surgical microscope[J].Progress in Modern Biomedicine,2012,12(16):3026-3028
[2]Starzl TE,Marchioro TL,Vonkaulla KN,et al.Homotransplantation of the liver in humans[J].Surgery,gynecology&obstetrics,1963,117:659-676
[3]Lee S,Charters AC,Chandler JG,et al.A technique for orthotopic liver transplantation in the rat[J].Transplantation,1973,16(6):664-669
[4]Kamada N,Calne RY.Orthotopic liver transplantation in the rat.Technique using cuff for portal vein anastomosis and biliary drainage[J].Transplantation,1979,28(1):47-50
[5]陈忠华,夏穗生.改进的大鼠原位肝移植术[J].武汉医学院学报,1984,05(04):244Chen Zhong-hua,Xia Shui-sheng.Modification of rat orthotopic liver transplatation model[J].Journal of Wu han Yi Xue Yuan,1984,05(04):244
[6]徐峰,杨甲梅,吴孟超,等.改良Kamada法大鼠原位肝移植模型的建立[J].中华肝胆外科杂志,2001,7(12):758-759Xu Feng,Yang Jia-mei,Wu Meng-chao,et al.Modification of Kamada technique in the rat model of orthotopic liver transplantation[J].Chinese Journal of Hepatobiliary Surgery,2001,7(12):758-759
[7]Nagai K,Yagi S,Uemoto S,et al.Surgical procedures for a rat model of partial orthotopic liver transplantation with hepatic arterial reconstruction[J].Journal of Visualized Experiments,2013,37(73):e4376
[8]Kakizaki Y,Miyagi S,Shimizu K,et al.The effects of short-term subnormothermic perfusion after cold preservation on liver grafts from donors after cardiac death:an ex vivo rat model[J].Transplantation,2018,15
[9]Chen X,Zheng J,Cai J,et al.The cytoskeleton proteinβ-actin may mediate T cell apoptosis during acute rejection reaction after liver transplantation in a rat model[J].Am J Transl Res,2017,9(11):4888-4901
[10]Shinya O,Tadahiro U,Zhao Xiang-dong,et al.Liver graft preservation using perfluorocarbon improves the outcomes of simulated donation after cardiac death liver transplantation in rats[J].Liver Transpl,2017,23(6):804-812
[11]马毅,何晓顺,陈规划.大鼠原位肝移植模型的手术技巧及并发症的预防[J].中华显微外科杂志,2003,26(1):45-47Ma Yi,He Xiao-shun,Chen Gui-hua.Surgical technique of the model of orthotopic liver transplantation and prevention of operational complication in rat[J].Chinese Journal of Microsurgery,2003,26(1):45-47
[12]Wang R,Shen Z,Yang L,et al.Protective effects of heme oxygenase-1-transduced bone marrow-derived mesenchymal stem cells on reduced size liver transplantation:role of autophagy regulated by the ERK/m TOR signaling pathway[J].Int J Mol Med,2017,40(5):1537-1548
[13]刘立,方成,王加谋.丙泊芬、戊巴比妥钠和乌拉坦对肾移植大鼠麻醉效果的比较[J].咸宁学院学报(医学版),2012,26(4):277-278Liu Li,Fang Cheng,Wang Jia-mou.Comparison on anaesthetic effect between propofol,sodium pentobarbital and urethane in rats with kidney transplantation[J].Journal of Xianning Univarsity(medical Sciences),2012,26(4):277-278
[14]陈辉,姜骊,王雁,等.三种不同麻醉药对SD大鼠肝移植手术麻醉效果的比较和应用[J].实验动物科学,2009,26(4):57-59Chen Hui,Jiang Li,Wang Yan,et al.Effect of three anesthetic drugs in SD rat liver transplantation surgery[J].Laboratory Animal Science,2009,26(4):57-59
[15]Cao D,Liu Y,Li J,et al.Isoflurane:an ideal anesthetic for rodent orthotopic liver transplantation surgery?[J].Transplant Proc,2016,48(8):2815-2820
[16]Luo C,Yuan D,Li X,et al.Propofol attenuated acute kidney injury after orthotopic liver transplantation via inhibiting gap junction composed of connexin 32[J].Anesthesiology,2015,122(1):72-86
[17]Schemmer P,Schoonhoven R,Swenberg JA,et al.Gentle in situ liver manipulation during organ harvest decreases survival after rat liver transplantation:role of Kupffer cells[J].Transplantation,1998,65(8):1015-1020
[18]Zeng XP,Li XJ,Zhang QY,et al.Tert-butylhydroquinone protects liver against ischemia reperfusion injury in rats through nrf2-activating anti-oxidative activity[J].Transplant Proc,2017,49(2):366-372
[19]Ishii E,Shimizu A,Takahashi M,et al.Surgical technique of orthotopic liver transplantation in rats:the Kamada technique and a new splint technique for hepatic artery reconstruction[J].Journal of Nippon Medical School,2013,80(1):4-15
[20]Zhi X,Xue F,Chen W,et al.OSI-027 modulates acute graft-versus-host disease after liver transplantation in a rat model[J].Liver Transpl,2017,23(9):1186-1198
[21]葛勇胜,许戈良,李建生,等.供肝的不同灌注方法在大鼠肝移植中的应用[J].实用医学杂志,2006,22(22):2585-2587Ge Yong-sheng,Xu Ge-liang,Li Jian-sheng,et al.Different hepatic perfusion procedures for donor liver transplantation[J].The Journal of PracticalMedicine,2006,22(22):2585-2587
[22]许赤,杨扬,易述红,等.稳定大鼠肝移植模型的规范及移植肝灌注方式比较[J].南方医科大学学报,2006,26(11):1556-1558Xu Chi,Yang Yang,Yi Shu-hong,et al.Standardization of rat stable orthotopic liver transplantation model and comparison of the effect of two liver graft perfusion methods[J].Journal of Southern Medical University,2006,26(11):1556-1558
[23]陈强星,李坤,孔伟浩,等.单人直视下改良建立大鼠原位肝移植模型[J].中华肝脏外科手术学电子杂志,2017,6(2):127-133Chen Qiang-xing,Li Kun,Kong Wei-hao,et al.Modi覱ed approach for establishment of rat models with orthotopic liver transplantation under direct vision of single operator[J].Chin J Hepat Surg(Electronic Edition),2017,6(2):127-133
[24]Czigany Z,Iwasaki J,Yagi S,et al.Improving research practice in rat orthotopic and partial orthotopic liver transplantation:a review,recommendation,and publication guide[J].European surgical research Europaische chirurgische Forschung Recherches chirurgicales europeennes,2015,55(1-2):119-138
[25]Yu E,Ueta H,Kimura H,et al.Graft-versus-host disease following liver transplantation:development of a high-incidence rat model and a selective prevention method[J].Am J Transplant,2017,17(4):979-991
[26]Ma B,Yang JY,Song WJ,et al.Combining Exosomes Derived from Immature DCs with donor antigen-specific treg cells induces tolerance in a rat liver allograft model[J].Sci Rep,2016,6:32971
[27]Tang J,Yang R,Lv L,et al.Transforming growth factor-β-expressing mesenchymal stem cells induce local tolerance in a rat liver transplantation model of acute rejection[J].Stem Cells,2016,34(11):2681-2692
[28]白建华,陈刚,朱新锋,等.原位肝移植模型的建立技巧与改进[J].中国组织工程研究,2015,19(40):6526-6530Bai Jian-hua,Chen Gang,Zhu Xin-feng,et al.Techniques and improvements of establishing orthotopic liver transplantation rat models[J].Chinese Journal of Tissue Engineering Research,2015,19(40):6526-6530
[29]Yoshimura N,Matsui S,Hamashima T,et al.The effects of perioperative portal venous inoculation with donor lymphocytes on renal allograft survival in the rat.I.specific prolongation of donor grafts and suppressor factor in the serum[J].Transplantation,1990,49(1):167-71
[30]许永刚,翁明哲,徐军明,等.单人建立大鼠原位肝移植模型的手术体会[J].现代生物医学进展,2012,12(16):3026-3028Xu Yong-gang,Weng Ming-zhe,Xu Jun-ming,et al.Technique of orthotopic liver transplantation in rats operated by one person without surgical microscope[J].Progress in Modern Biomedicine,2012,12(16):3026-3028