Expression of Transforming Growth Factor-β1 in Liver Tissue of Acute Rejection after Liver Transplantation in Rhesus_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 RANJiang-hua , ZHANGXi-bing , LIUJing , ZHANGSheng-ning , LILai-bang , CHENYi-ming , GAOYang , LIWang ,  LILi

Department of Hepatobiliary and Pancreatic Surgery, Affiliated Ganmei Hospital of Kunming Medical University, Liver Transplantation Center of Organ Transplantation Institute of Yunnan Province, Kunming 650011, Yunnan Province, China;

Corresponding author：

LILi, Email: ynkmlili62@hotmail.com

Keywords：

Rhesus; Liver transplantation; Acute rejection; Transforming growth factor-β1; Immunohistoche-mistry method; Western blot method

DOI：

10.7507/1007-9424.20140331

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Objective To investigate the early diagnostic value of transforming growth factor-β1(TGF-β1) on acute rejection after liver transplantation in rhesus by detecting the expression of TGF-β1 in the liver tissue. Methods Liver transplantation models in rhesus were constructed by the improved vascular dual cuff, supporting tube of biliary tract, and artery anastomosis method.The successful models were randomly divided into experimental group (no immunosuppressant treatment in perioperative period) and control group (treated by immunosuppressant in perioperative period).Then the blood samples and liver tissues were collected at 6, 12, 24, and 72 hours after surgery.Allograft rejections of liver tissue after liver transplantation were monitored by liver function test, hematoxylin-eosin staining and Banff score.Finally, the expression level of TGF-β1 was detected by Western blot analysis or immunohistochemistry technique. Results ①The acute rejection happened in all the rhesus at 12 h, 24 h and 72 h after liver transplantation, especially at 72 h after liver transplantation in the experimental group, the Banff grade levels of acute rejection in the liver tissue was more severe than that in the control group (P < 0.05).②The levels of ALT, AST, and TBIL after liver transplantation was gradually increased, which were similar at 6 h and 12 h after transplantation between the two groups, but which at 24 h and 72 h after transplantation in the experimental group were significantly higher than those in the control group (P < 0.05).③The results of TGF-β1 protein expression using immunohistochemical detection:The percentage of positive area of TGF-β1 of liver tissue at 12 h in the experimental group was significantly higher than that in the control group (P < 0.05).With the extension of time, it was gradually increased and significantly higher than that in the control group at 24 h or 72 h (P < 0.05).④The semi-quantitative results of TGF-β1 protein expression using Western blot detection:The TGF-β1 protein expressions began to increase at 6 h after liver transplantation in the experimental group and the control group, and the magnitude of increase was more obvious in the experimental group.The TGF-β1 protein expressions at different time (6 h, 12 h, 24 h, and 72 h) in the experimental group were significantly higher than those in the control group (P value was 0.003, 0.001, 0.001, and 0.001, respectively). Conclusions The elevated level of TGF-β1 of liver tissue after liver transplantation might suggest the enhanced cellular immune function, it might have certain significance for early diagnosis of acute rejection after liver transplantation.

Citation： RANJiang-hua, ZHANGXi-bing, LIUJing, ZHANGSheng-ning, LILai-bang, CHENYi-ming, GAOYang, LIWang, LILi. Expression of Transforming Growth Factor-β1 in Liver Tissue of Acute Rejection after Liver Transplantation in Rhesus. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2014, 21(11): 1384-1390. doi: 10.7507/1007-9424.20140331 Copy

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2.	Rook M, Rand E.Predictors of long-term outcome after liver transplant[J].Curr Opin Organ Transplant, 2011, 16(5):499-504.
3.	Tambur AR, Ortegel JW, Ben-Ari Z, et al.Role of cytokine gene polymorphism in hepatitis C recurrence and allograft rejection among liver transplant recipients[J].Transplantation, 2001, 71(10):1475-1480.
4.	Morris-Stiff G.TGFbeta-1 and the development of chronic graft nephropathy:relative roles of gene, mRNA and protein[J].Ann R Coll Surg Engl, 2005, 87(5):326-330.
5.	Hoffmann S, Park J, Jacobson LM, et al.Donor genomics influence graft events:the effect of donor polymorphisms on acute rejection and chronic allograft nephropathy[J].Kidney Int, 2004, 66(4):1686-1693.
6.	Di Filippo S, Zeevi A, McDade KK, et al.Impact of TGFbeta1 gene polymorphisms on acute and chronic rejection in pediatric heart transplant allografts[J].Transplantation, 2006, 81(6):934-939.
7.	中华人民共和国科学技术部.关于发布《关于善待实验动物的指导性意见》的通知[EB/OL].http://www.most.gov.cn/fggw/zfwj/zfwj2006/zf06wj/zf06bfw/200609/t20060930_54196.htm.
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10.	Busuttil RW, Lake JR.Role of tacrolimus in the evolution of liver transplantation[J].Transplantation, 2004, 77(9 suppl):S44-S51.
11.	Solez K.History of the Banff classification of allograft pathology as it approaches its 20th year[J].Curr Opin Organ Transplant, 2010, 15(1):49-51.
12.	Racusen LC, Halloran PF, Solez K.Banff 2003 meeting report:new diagnostic insights and standards[J].Am J Transplant, 2004, 4(10):1562-1566.
13.	Rüdiger T, Höfler H, Kreipe HH, et al.Quality assurance in immunohistochemistry:results of an interlaboratory trial involving 172 pathologists[J].Am J Surg Pathol, 2002, 26(7):873-882.
14.	Dickson C.Protein techniques:immunoprecipitation, in vitro kinase assays, and Western blotting[J].Methods Mol Biol, 2008, 461:735-744.
15.	Opelz G, Döhler B, Collaborative Transplant Study Report.Influence of time of rejection on long-term graft survival in renal transplantation[J].Transplantation, 2008, 85(5):661-666.
16.	Stehlik J, Edwards LB, Kucheryavaya AY, et al.The registry of the international society for heart and lung transplantation:29th official adult heart transplant report-2012[J].J Heart Lung Transplant, 2012, 31(10):1052-1064.
17.	Thurairajah PH, Carbone M, Bridgestock H, et al.Late acute liver allograft rejection; a study of its natural history and graft survival in the current era[J].Transplantation, 2013, 95(7):955-959.
18.	Geissmann F, Revy P, Regnault A, et al.TGF-beta 1 prevents the noncognate maturation of human dendritic Langerhans cells[J].J Immunol, 1999, 162(8):4567-4575.
19.	Ohtani T, Mizuashi M, Nakagawa S, et al.TGF-beta1 dampens the susceptibility of dendritic cells to environmental stimulation, leading to the requirement for danger signals for activation[J].Immunology, 2009, 126(4):485-499.
20.	Mollah ZU, Aiba S, Nakagawa S, et al.Interleukin-3 in cooperation with transforming growth factor beta induces granulocyte macrophage colony stimulating factor Independent differentiation of human CD34⁺ hematopoietic progenitor cells into dendritic cells with features of Langerhans cells[J].J Invest Dermatol, 2003, 121(6):1397-1401.
21.	Border WA, Ruoslahti E.Transforming growth factor-beta in disease:the dark side of tissue repair[J].J Clin Invest, 1992, 90(1):1-7.
22.	Böttinger EP.TGF-βin renal injury and disease[J].Semin Nephrol, 2007, 27(3):309-320.
23.	Rifkin DB, Kojima S, Abe M, et al.TGF-beta:structure, function, and formation[J].Thromb Haemost, 1993, 70(1):177-179.
24.	Flaumenhaft R, Kojima S, Abe M, et al.Activation of latent transforming growth factor beta[J].Adv Pharmacol, 1993, 24:51-76.
25.	Hutchinson IV.The role of transforming growth factor-beta in transplant rejection[J].Transplant Proc, 1999, 31(7A):9S-13S.

1. Alex Bishop G, Bertolino PD, Bowen DG, et al.Tolerance in liver transplantation[J].Best Pract Res Clin Gastroenterol, 2012, 26(1):73-84.
2. Rook M, Rand E.Predictors of long-term outcome after liver transplant[J].Curr Opin Organ Transplant, 2011, 16(5):499-504.
3. Tambur AR, Ortegel JW, Ben-Ari Z, et al.Role of cytokine gene polymorphism in hepatitis C recurrence and allograft rejection among liver transplant recipients[J].Transplantation, 2001, 71(10):1475-1480.
4. Morris-Stiff G.TGFbeta-1 and the development of chronic graft nephropathy:relative roles of gene, mRNA and protein[J].Ann R Coll Surg Engl, 2005, 87(5):326-330.
5. Hoffmann S, Park J, Jacobson LM, et al.Donor genomics influence graft events:the effect of donor polymorphisms on acute rejection and chronic allograft nephropathy[J].Kidney Int, 2004, 66(4):1686-1693.
6. Di Filippo S, Zeevi A, McDade KK, et al.Impact of TGFbeta1 gene polymorphisms on acute and chronic rejection in pediatric heart transplant allografts[J].Transplantation, 2006, 81(6):934-939.
7. 中华人民共和国科学技术部.关于发布《关于善待实验动物的指导性意见》的通知[EB/OL].http://www.most.gov.cn/fggw/zfwj/zfwj2006/zf06wj/zf06bfw/200609/t20060930_54196.htm.
8. 冉江华, 张升宁, 刘静, 等.恒河猴同种异体原位肝移模型的改进[J].中国普外基础与临床杂志, 2010, 17(7):694-698.
9. 冉江华, 李铸, 刘静, 等.二袖套法建立恒河猴原位肝移植的稳定模型[J].中国组织工程研究与临床康复, 2011, 15(5):762-768.
10. Busuttil RW, Lake JR.Role of tacrolimus in the evolution of liver transplantation[J].Transplantation, 2004, 77(9 suppl):S44-S51.
11. Solez K.History of the Banff classification of allograft pathology as it approaches its 20th year[J].Curr Opin Organ Transplant, 2010, 15(1):49-51.
12. Racusen LC, Halloran PF, Solez K.Banff 2003 meeting report:new diagnostic insights and standards[J].Am J Transplant, 2004, 4(10):1562-1566.
13. Rüdiger T, Höfler H, Kreipe HH, et al.Quality assurance in immunohistochemistry:results of an interlaboratory trial involving 172 pathologists[J].Am J Surg Pathol, 2002, 26(7):873-882.
14. Dickson C.Protein techniques:immunoprecipitation, in vitro kinase assays, and Western blotting[J].Methods Mol Biol, 2008, 461:735-744.
15. Opelz G, Döhler B, Collaborative Transplant Study Report.Influence of time of rejection on long-term graft survival in renal transplantation[J].Transplantation, 2008, 85(5):661-666.
16. Stehlik J, Edwards LB, Kucheryavaya AY, et al.The registry of the international society for heart and lung transplantation:29th official adult heart transplant report-2012[J].J Heart Lung Transplant, 2012, 31(10):1052-1064.
17. Thurairajah PH, Carbone M, Bridgestock H, et al.Late acute liver allograft rejection; a study of its natural history and graft survival in the current era[J].Transplantation, 2013, 95(7):955-959.
18. Geissmann F, Revy P, Regnault A, et al.TGF-beta 1 prevents the noncognate maturation of human dendritic Langerhans cells[J].J Immunol, 1999, 162(8):4567-4575.
19. Ohtani T, Mizuashi M, Nakagawa S, et al.TGF-beta1 dampens the susceptibility of dendritic cells to environmental stimulation, leading to the requirement for danger signals for activation[J].Immunology, 2009, 126(4):485-499.
20. Mollah ZU, Aiba S, Nakagawa S, et al.Interleukin-3 in cooperation with transforming growth factor beta induces granulocyte macrophage colony stimulating factor Independent differentiation of human CD34⁺ hematopoietic progenitor cells into dendritic cells with features of Langerhans cells[J].J Invest Dermatol, 2003, 121(6):1397-1401.
21. Border WA, Ruoslahti E.Transforming growth factor-beta in disease:the dark side of tissue repair[J].J Clin Invest, 1992, 90(1):1-7.
22. Böttinger EP.TGF-βin renal injury and disease[J].Semin Nephrol, 2007, 27(3):309-320.
23. Rifkin DB, Kojima S, Abe M, et al.TGF-beta:structure, function, and formation[J].Thromb Haemost, 1993, 70(1):177-179.
24. Flaumenhaft R, Kojima S, Abe M, et al.Activation of latent transforming growth factor beta[J].Adv Pharmacol, 1993, 24:51-76.
25. Hutchinson IV.The role of transforming growth factor-beta in transplant rejection[J].Transplant Proc, 1999, 31(7A):9S-13S.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Expression of Transforming Growth Factor-β1 in Liver Tissue of Acute Rejection after Liver Transplantation in Rhesus

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