• 1. 四川大學華西醫(yī)院肝移植中心（成都 610041）;;
  • 2. 成都地奧集團公司新藥部（成都 610041）;

目的　 探討鋅指蛋白A20對同種異體大鼠30%小體積移植肝再生與排斥及受體存活時間的影響。
方法 　采用急性排斥組合DA (RT1a)大鼠→Lewis (RT1l)大鼠建立同種異體大鼠30%小體積肝移植模型。 75只大鼠均分為A20腺病毒組（A20組）、對照空腺病毒組（rAdEasy組）及對照生理鹽水組（PS組）3組。 供肝切取后采用離體門靜脈灌注法轉(zhuǎn)基因干預(yù)，肝臟灌洗后移植。 觀察受體存活時間及排斥反應(yīng)，檢測移植肝A20表達、肝細胞再生與凋亡、肝血竇內(nèi)皮細胞(LSECs)中NF-κB活性與ICAM-1 mRNA表達、移植肝浸潤單核細胞（LIMCs）總數(shù)及自然殺傷(NK)細胞和自然殺傷T（NKT）細胞數(shù)以及血清IFN-γ 水平。
結(jié)果 　A20組受體大鼠存活時間長于PS組大鼠和rAdEasy組大鼠(P=0.001 8)，而PS組大鼠存活時間又長于rAdEasy組大鼠(P=0.001 8)。 A20促進小體積移植肝再生，肝移植后4 d A20組大鼠肝細胞BrdU標記指數(shù)高于PS組大鼠和rAdEasy組大鼠(P＜0.01)； A20組大鼠肝細胞凋亡指數(shù)低于PS組大鼠和rAdEasy組大鼠(P＜0.01)。 肝移植術(shù)后4 d，組織學檢測結(jié)果顯示A20組大鼠移植肝有少量細胞浸潤，呈輕度排斥； 而PS 組和rAdEasy組大鼠移植肝有大量細胞浸潤，呈重度排斥。 A20能抑制移植后早期(1 d) LSECs 中NF-κB活性和 ICAM-1 mRNA 表達。 流式細胞術(shù)檢測結(jié)果顯示A20能下調(diào)移植肝LIMCs數(shù)，包括下調(diào)NK和NKT 亞群細胞數(shù)，并下調(diào)血清IFN-γ 水平(P＜0.05)。
結(jié)論 　A20能有效促進同種異體大鼠30%小體積移植肝再生，抑制排斥并延長受體存活時間，其功效可能與抑制LSECs中NF-κB活性、抑制LIMCs浸潤及減少NK 細胞和NKT 細胞浸潤入肝以及抑制肝細胞凋亡有關(guān)。

引用本文： 徐明清,嚴律南,茍興華,彭方興,李德華,韓蕾,黃蘭英,黃迎春,胡海洋. 鋅指蛋白A20促進同種異體大鼠小體積移植肝再生并抑制急性排斥. 中國普外基礎(chǔ)與臨床雜志, 2008, 15(8): 572-578. doi: 復(fù)制

1. 　Omura T, Nakagawa T, Randall HB, et al. Increased immune responses to regenerating partial liver grafts in the rat [Ｊ]. J Surg Res, 1997; 70(1)∶ 34.
2. 　Shiraishi M, Csete ME, Yasunaga C, et al. Regeneration-induced accelerated rejection in reduced-size liver grafts [Ｊ]. Transplantation, 1994； 57(3)∶ 336.
3. 　Huda KA, Guo L, Haga S, et al. Ex vivo adenoviral gene transfer of constitutively activated STAT3 reduces post-transplant liver injury and promotes regeneration in a 20% rat partial liver transplant model [Ｊ]. Transplant Int， 2006; 19(5)∶ 415.
4. 　Braet F, De Zanger R, Sasaoki T, et al. Assessment of a method of isolation, purification and cultivation of rat liver sinusoidal endothelial cells [Ｊ]. Lab Invest, 1994; 70(6)∶ 944.
5. 　Ogura Y, Martinez OM, Villanueva JC, et al. Apoptosis and allograft rejection in the absence of CD8+ T cells [Ｊ]. Transplantation, 2001; 71(12)∶ 1827.
6. 　Xu MQ, Suo YP, Gong JP, et al. Augmented regeneration of partial liver allograft induced by nuclear factor-κB decoy oligodeoxynucleotides-modified dendritic cells [Ｊ]. World J Gastroentrol, 2004； 10(4)∶ 573.
7. 　Luo D, Vanderkerken K, Bouwens L, et al. The role of adhesion molecules in the recruitment of hepatic natural killer cells (pit cells) in rat liver [Ｊ]. Hepatology, 1996; 24(6)∶ 1475.
8. 　Adams DH， Hubscher SG， Shaw J, et al. Intercellular adhesion molecule-1 on liver allografts during rejection [Ｊ]. Lancet， 1989; 2(8672)∶ 1122.
9. 　Steinhoff G, Behrend M, Schrader B, et al. Intercellular immune adhesion molecules in human liver transplants: Overview on expression patterns of leukocyte receptor and ligand molecules [Ｊ]. Hepatology, 1993; 18(2)∶ 440.
10. 　Omura T, Ishikura H, Nakajima Y, et al. The expression of LFA-1/ICAM-1 in liver transplantation in rats [Ｊ]. Transplant Proc, 1992; 24(4)∶ 1618.
11. 　Opipari AW Jr, Boguski MS, Dixit VM. The A20 cDNA induced by tumor necrosis factor alpha encodes a novel type of zinc finger protein [Ｊ]. J Biol Chem, 1990; 265(25)∶ 14705.
12. 　Cooper JT, Stroka DM, Brostjan C, et al. A20 blocks endothelial cell activation through a NF-κB-dependent mechanism [Ｊ]. J Biol Chem, 1996; 271(30)∶ 18068.
13. 　Ferran C, Stroka DM, Badrichani AZ, et al. A20 inhibits NF-κB activation in endothelial cells without sensitizing to tumor necrosis factor-mediated apoptosis [Ｊ]. Blood, 1998; 91(7)∶ 2249.
14. 　Longo CR, Arvelo MB, Patel VI, et al. A20 protects from CD40-CD40 ligand-mediated endothelial cell activation and apoptosis [Ｊ]. Circulation, 2003; 108(9)∶ 1113.
15. 　Arvelo MB, Cooper JT, Longo C, et al. A20 protects mice from galactosmine/lipopolysaccharide acute toxic lethal hepatitis [Ｊ]. Hepatology, 2002; 35(3)∶ 535.
16. 　Hoffmann MW， Wonigeit K, Steinhoff G, et al. Production of cytokines (TNF-alpha, IL-1-beta) and endothelial cell activation in human liver allograft rejection [Ｊ]. Transplantation, 1993; 55(2)∶ 329.
17. 　Tamura F, Masuhara A, Sakaida I, et al. FK506 promotes liver regeneration by suppressing natural killer cell activity [Ｊ]. J Gastroenterol Hepatol， 1998; 13(7)∶ 703.
18. 　Huang W, Dong Z, Wei H, et al. Selective elimination of hepatic natural killer T cells with concanavalin A improves liver regeneration in mice [Ｊ]. Liver Int， 2006; 26(3)∶ 339.
19. 　Sato Y, Tsukada K, Matsumoto Y, et al. Interferon-gamma inhibits liver regeneration by stimulating major histocompatibility complex class II antigen expression by regenerating liver [Ｊ]. Hepatology, 1993; 18(2)∶ 340.
20. 　Obara H, Nagasaki K, Hsieh CL, et al. IFN-γ, produced by NK cells that infiltrate liver allografts early after transplantation, links the innate and adaptive immune responses [Ｊ]. Am J Transplant, 2005; 5(9)∶ 2094.
21. 　Navarro F, Portalès P, Candon S, et al. Natural killer cell and αβ and γδ lymphocyte traffic into the liver graft immediately after liver transplantation [Ｊ]. Transplantation, 2000; 69(4)∶〖KG-8〗 633.
22. 　Longo CR, Patel VI, Shrikhande GV, et al. A20 protects mice from lethal radical hepatectomy by promoting hepatocyte proliferation via a p21waf1-dependent mechanism [Ｊ]. Hepatology, 2005; 42(1)∶ 156.
23. 　Yamamoto H, Ohdan H, Shintaku S, et al. Role of the bcl-2/bax pathway in hepatocyte apoptosis during acute rejection after rat liver transplantation [Ｊ]. Transpl Int， 1998; 11 (Suppl 1)∶ S179.
24. 　Krams SM, Egawa H, Quinn MB, et al. Apoptosis as a mechanism of cell death in liver allograft rejection [Ｊ]. Transplantation, 1995; 59(4)∶ 621.
25. 　Hsieh CL, Obara H, Ogura Y, et al. NK cells and transplantation [Ｊ]. Transpl Immunol， 2002; 9(2-4)∶ 111.
26. 　Kojima Y, Kawasaki-Koyanagi A, Sueyoshi N, et al. Localization of Fas ligand in cytoplasmic granules of CD8+ cytotoxic T lymphocytes and natural killer cells： participation of Fas ligand in granule exocytosis model of cytotoxicity [Ｊ]. Biochem Biophys Res Commun， 2002; 296(2)∶ 328.
27. 　Ariki N, Morimoto Y, Yagi T, et al. Activated T cells and soluble molecules in the portal venous blood of patients with cholestatic and hepatitis C virus-positive liver cirrhosis. Possible promotion of Fas/FasL-mediated apoptosis in the bile-duct cells and hepatocyte injury [Ｊ]. J Int Med Res， 2003; 31(3)∶ 170.
28. 　Wang J, Li W, Min J, et al. Fas siRNA reduces apoptotic cell death of allogeneic-transplanted hepatocytes in mouse spleen [Ｊ]. Transplant Proc， 2003; 35(4)∶ 1594.
  1. 1. 　Omura T, Nakagawa T, Randall HB, et al. Increased immune responses to regenerating partial liver grafts in the rat [Ｊ]. J Surg Res, 1997; 70(1)∶ 34.
  2. 2. 　Shiraishi M, Csete ME, Yasunaga C, et al. Regeneration-induced accelerated rejection in reduced-size liver grafts [Ｊ]. Transplantation, 1994； 57(3)∶ 336.
  3. 3. 　Huda KA, Guo L, Haga S, et al. Ex vivo adenoviral gene transfer of constitutively activated STAT3 reduces post-transplant liver injury and promotes regeneration in a 20% rat partial liver transplant model [Ｊ]. Transplant Int， 2006; 19(5)∶ 415.
  4. 4. 　Braet F, De Zanger R, Sasaoki T, et al. Assessment of a method of isolation, purification and cultivation of rat liver sinusoidal endothelial cells [Ｊ]. Lab Invest, 1994; 70(6)∶ 944.
  5. 5. 　Ogura Y, Martinez OM, Villanueva JC, et al. Apoptosis and allograft rejection in the absence of CD8+ T cells [Ｊ]. Transplantation, 2001; 71(12)∶ 1827.
  6. 6. 　Xu MQ, Suo YP, Gong JP, et al. Augmented regeneration of partial liver allograft induced by nuclear factor-κB decoy oligodeoxynucleotides-modified dendritic cells [Ｊ]. World J Gastroentrol, 2004； 10(4)∶ 573.
  7. 7. 　Luo D, Vanderkerken K, Bouwens L, et al. The role of adhesion molecules in the recruitment of hepatic natural killer cells (pit cells) in rat liver [Ｊ]. Hepatology, 1996; 24(6)∶ 1475.
  8. 8. 　Adams DH， Hubscher SG， Shaw J, et al. Intercellular adhesion molecule-1 on liver allografts during rejection [Ｊ]. Lancet， 1989; 2(8672)∶ 1122.
  9. 9. 　Steinhoff G, Behrend M, Schrader B, et al. Intercellular immune adhesion molecules in human liver transplants: Overview on expression patterns of leukocyte receptor and ligand molecules [Ｊ]. Hepatology, 1993; 18(2)∶ 440.
  10. 10. 　Omura T, Ishikura H, Nakajima Y, et al. The expression of LFA-1/ICAM-1 in liver transplantation in rats [Ｊ]. Transplant Proc, 1992; 24(4)∶ 1618.
  11. 11. 　Opipari AW Jr, Boguski MS, Dixit VM. The A20 cDNA induced by tumor necrosis factor alpha encodes a novel type of zinc finger protein [Ｊ]. J Biol Chem, 1990; 265(25)∶ 14705.
  12. 12. 　Cooper JT, Stroka DM, Brostjan C, et al. A20 blocks endothelial cell activation through a NF-κB-dependent mechanism [Ｊ]. J Biol Chem, 1996; 271(30)∶ 18068.
  13. 13. 　Ferran C, Stroka DM, Badrichani AZ, et al. A20 inhibits NF-κB activation in endothelial cells without sensitizing to tumor necrosis factor-mediated apoptosis [Ｊ]. Blood, 1998; 91(7)∶ 2249.
  14. 14. 　Longo CR, Arvelo MB, Patel VI, et al. A20 protects from CD40-CD40 ligand-mediated endothelial cell activation and apoptosis [Ｊ]. Circulation, 2003; 108(9)∶ 1113.
  15. 15. 　Arvelo MB, Cooper JT, Longo C, et al. A20 protects mice from galactosmine/lipopolysaccharide acute toxic lethal hepatitis [Ｊ]. Hepatology, 2002; 35(3)∶ 535.
  16. 16. 　Hoffmann MW， Wonigeit K, Steinhoff G, et al. Production of cytokines (TNF-alpha, IL-1-beta) and endothelial cell activation in human liver allograft rejection [Ｊ]. Transplantation, 1993; 55(2)∶ 329.
  17. 17. 　Tamura F, Masuhara A, Sakaida I, et al. FK506 promotes liver regeneration by suppressing natural killer cell activity [Ｊ]. J Gastroenterol Hepatol， 1998; 13(7)∶ 703.
  18. 18. 　Huang W, Dong Z, Wei H, et al. Selective elimination of hepatic natural killer T cells with concanavalin A improves liver regeneration in mice [Ｊ]. Liver Int， 2006; 26(3)∶ 339.
  19. 19. 　Sato Y, Tsukada K, Matsumoto Y, et al. Interferon-gamma inhibits liver regeneration by stimulating major histocompatibility complex class II antigen expression by regenerating liver [Ｊ]. Hepatology, 1993; 18(2)∶ 340.
  20. 20. 　Obara H, Nagasaki K, Hsieh CL, et al. IFN-γ, produced by NK cells that infiltrate liver allografts early after transplantation, links the innate and adaptive immune responses [Ｊ]. Am J Transplant, 2005; 5(9)∶ 2094.
  21. 21. 　Navarro F, Portalès P, Candon S, et al. Natural killer cell and αβ and γδ lymphocyte traffic into the liver graft immediately after liver transplantation [Ｊ]. Transplantation, 2000; 69(4)∶〖KG-8〗 633.
  22. 22. 　Longo CR, Patel VI, Shrikhande GV, et al. A20 protects mice from lethal radical hepatectomy by promoting hepatocyte proliferation via a p21waf1-dependent mechanism [Ｊ]. Hepatology, 2005; 42(1)∶ 156.
  23. 23. 　Yamamoto H, Ohdan H, Shintaku S, et al. Role of the bcl-2/bax pathway in hepatocyte apoptosis during acute rejection after rat liver transplantation [Ｊ]. Transpl Int， 1998; 11 (Suppl 1)∶ S179.
  24. 24. 　Krams SM, Egawa H, Quinn MB, et al. Apoptosis as a mechanism of cell death in liver allograft rejection [Ｊ]. Transplantation, 1995; 59(4)∶ 621.
  25. 25. 　Hsieh CL, Obara H, Ogura Y, et al. NK cells and transplantation [Ｊ]. Transpl Immunol， 2002; 9(2-4)∶ 111.
  26. 26. 　Kojima Y, Kawasaki-Koyanagi A, Sueyoshi N, et al. Localization of Fas ligand in cytoplasmic granules of CD8+ cytotoxic T lymphocytes and natural killer cells： participation of Fas ligand in granule exocytosis model of cytotoxicity [Ｊ]. Biochem Biophys Res Commun， 2002; 296(2)∶ 328.
  27. 27. 　Ariki N, Morimoto Y, Yagi T, et al. Activated T cells and soluble molecules in the portal venous blood of patients with cholestatic and hepatitis C virus-positive liver cirrhosis. Possible promotion of Fas/FasL-mediated apoptosis in the bile-duct cells and hepatocyte injury [Ｊ]. J Int Med Res， 2003; 31(3)∶ 170.
  28. 28. 　Wang J, Li W, Min J, et al. Fas siRNA reduces apoptotic cell death of allogeneic-transplanted hepatocytes in mouse spleen [Ｊ]. Transplant Proc， 2003; 35(4)∶ 1594.