• 1.首都醫(yī)科大學(xué)宣武醫(yī)院普外科(北京 100053);;
  • 2.首都醫(yī)科大學(xué)宣武醫(yī)院血液科流式細(xì)胞室(北京 100053);;
  • 3.首都醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院人體解剖與組織胚胎學(xué)系(北京 100054);

目的  探討雷公藤內(nèi)酯醇(triptolide,TPT)在小鼠同種異體胰島移植中的抗排斥作用及其機(jī)理。
方法  采用BALB/c小鼠作為供體,進(jìn)行胰島分離,以鏈脲佐菌素(streptozotocin,STZ)誘導(dǎo)的C57BL/6糖尿病小鼠作為受體,行左腎被膜下胰島移植。將移植后糖尿病小鼠隨機(jī)(隨機(jī)數(shù)字表法)分為3組,每組8只。于胰島移植術(shù)后前5d分別給予腹腔注射1%吐溫80溶劑(對照組)、TPT 50μg/kg (L-TPT組)和100μg/kg (H-TPT組),之后隔天注射1次,至術(shù)后第14天結(jié)束。術(shù)后監(jiān)測受體血糖水平變化; 并于術(shù)后第10天每組隨機(jī)(隨機(jī)數(shù)字表法)選取3只小鼠,切取左側(cè)腎臟行病理學(xué)檢查,流式細(xì)胞術(shù)檢測脾淋巴細(xì)胞中CD4+CD25+Foxp3+調(diào)節(jié)性 T細(xì)胞比例。
結(jié)果  對照組、L-TPT組和H-TPT組移植胰島的中位存活時(shí)間分別為12.6d (9 ~16d)、21.4d (14~27d)和27.6d (19~34d);脾淋巴細(xì)胞中CD4+CD25+Foxp3+調(diào)節(jié)性 T細(xì)胞比例分別為(5.2±0.6) %、(12.0±1.3)%和(15.7±1.8)%。與對照組相比,L-TPT組和H-TPT組小鼠移植胰島的中位存活時(shí)間明顯延長(P<0.05),CD4+CD25+Foxp3+調(diào)節(jié)性T細(xì)胞比例顯著增高(P<0.05)。
結(jié)論  TPT通過上調(diào)移植受體CD4+CD25+Foxp3+調(diào)節(jié)性T細(xì)胞比例,減輕了胰島移植后的排斥反應(yīng),顯著延長了移植胰島的存活時(shí)間,其免疫抑制作用呈劑量依賴性。

引用本文: 梁闊,劉爽,崔葉青,孫海晨,羅斌,萬歲桂,孫雪靜,郭多,李非. 雷公藤內(nèi)酯醇在小鼠同種異體胰島移植中的抗排斥作用△. 中國普外基礎(chǔ)與臨床雜志, 2013, 20(7): 737-741. doi: 復(fù)制

1. Shapiro AM, Lakey JR, Ryan EA, et al. Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen[J]. N Engl J Med, 2000, 343(4):230-238.
2. Ryan EA, Paty BW, Senior PA, et al. Five-year follow-up after clinical islet transplantation[J]. Diabetes, 2005, 54(7):2060-2069.
3. Larsen JL, Bennett RG, Burkman T, et al. Tacrolimus and sirolimus cause insulin resistance in normal sprague dawley rats[J]. Transplantation, 2006, 82(4): 466-470.
4. Wang J, Xu R, Jin R, et al. Immunosuppressive activity of the Chinese medicinal plant Tripterygium wilfordii. Ⅰ. Prolongation of rat cardiac and renal allograft survival by the PG27 extract and immunosuppressive synergy in combination therapy with cyclosporine[J]. Transplantation, 2000, 70(3): 447-455.
5. Li R, Takazawa K, Suzuki H, et al. Synergistic effect of triptolideand tacrolimus on rat cardiac allotransplantation[J]. Jpn Heart J, 2004, 45(4):657-665.
6. Liu Y, Chen Y, Liu FQ, et al. Combined treatment with triptolideand rapamycin prolongs graft survival in a mouse model of cardiactransplantation[J]. Transpl Int, 2008, 21(5):483-494.
7. Zhang G, Liu Y, Guo H, et al. Triptolide promotes generation of FoxP3+ T regulatory cells in rats[J]. J Ethnopharmacol, 2009, 125(1): 41-46.
8. 鄭樹森, 郭華, 俞軍, 等. 雷公藤多甙抑制大鼠異種肝移植急性排斥的研究[J]. 中華器官移植雜志, 2003, 24 (6):223-225.
9. 劉爽, 崔士華, 孫海晨, 等. 一種改良的小鼠胰島分離和移植模型[J]. 首都醫(yī)科大學(xué)學(xué)報(bào), 2009, 30(2):259-261.
10. Xin MJ, Cui SH, Liu S, et al. Triptolide prolonged allogeneic islet graft survival in chemically induced and spontaneously diabeticmice without impairment of islet function[J]. Hepatobiliary Pancreat Dis Int, 2010, 9(3):312-318.
11. Liang K, Du W, Zhu W, et al. Contribution of different mecha-nisms to pancreatic beta-cell hyper-secretion in non-obese diabetic(NOD) mice during pre-diabetes[J]. J Biol Chem, 2011, 286(45):39537-39545.
12. 徐澤寬, 苗毅. 胰腺移植和胰島移植的現(xiàn)狀[J]. 中國普外基礎(chǔ)與臨床雜志, 2003, 10(6): 532-534.
13. 張劍, 陳規(guī)劃, 翁建平, 等. 抗CD40L 單克隆抗體在異種胰島移植排斥反應(yīng)中的作用[J]. 中國普外基礎(chǔ)與臨床雜志, 2008, 15(5): 329-332.
14. 李柏峰, 劉永鋒, 程穎, 等. 氨基胍對大鼠胰島保護(hù)作用的實(shí)驗(yàn)研究[J]. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(9): 729-733.
15. Lo DJ, Anderson DJ, Weaver TA, et al. Belatacept and sirolimusprolong nonhuman primate renal allograft survival without arequirement for memory T cell depletion[J]. Am J Transplant, 2013, 13(2): 320-328.
16. Lowe M, Badell IR, Thompson P, et al. A novel monoclonal antibody to CD40 prolongs islet allograft survival[J]. Am J Transplant, 2012, 12(8): 2079-2087.
17. Yin N, Zhang N, Xu J, et al. Targeting lymphangiogenesis after islet transplantation prolongs islet allograft survival[J]. Transplantation, 2011, 92(1) :25-30.
18. Nataraju A, Saini D, Ramachandran S, et al. Oleanolic acid, a plant triterpenoid, significantly improves survival and function of islet allograft[J]. Transplantation, 2009, 88(8):987-994.
19. Zhou R, Zhang F, He PL, et al. (5R)-5-hydroxytriptolide (LLDT-8), a novel triptolide analog mediates immunosuppressive effects in vitro and in vivo[J]. Int Immunopharmacol, 2005, 5(13-14):1895-1903.
20. Liu Q, Chen T, Chen G, et al. Triptolide impairs dendritic cell migration by inhibiting CCR7 and COX-2 expression through PI3-K/Akt and NF-kappaB pathways[J]. Mol Immunol, 2007,44(10): 2686-2696.
21. Hong Y, Zhou W, Li K, et al. Triptolide is a potent suppressant of C3, CD40 and B7h expression in activated human proximaltubular epithelial cells[J]. Kidney Int, 2002, 62(4):1291-1300.
22. Liu Y, Chen Y, Lamb JR, et al. Triptolide, a component of Chinese herbal medicine, modulates the functional phenotype of dendritic cells[J]. Transplantation, 2007, 84(11): 1517-1526.
23. Hoffmann P, Ermann J, Edinger M, et al. Donor-type CD4+CD25+T regulatory T cells suppress lethal acute graft-versus-host disease after allogeneic bone marrow transplantation[J]. J Exp Med, 2002, 196(3): 389-399.
24. Nguyen VH , Shashidhar S, Chang DS, et al. The impact of regulatory T cells on T-cell immunity following hematopoietic cell transplantation[J] . Blood, 2008, 111(2): 945- 953.
25. Jiang X, Sun W, Guo D, et al. Cardiac allograft acceptance induced by blockade of CD40-CD40L costimulation is dependent on CD4+CD25+regulatory T cells[J] . Surgery, 2011, 149(3):336-346.
26. Sakaguchi S. Naturally arising CD4+ regulatory T cells for immu-nologic self tolerance and negative control of immune responses[J]. Annu Rev Immunol, 2004, 22: 531-562.
27. Shevach EM, McHugh RS, Piccirillo CA, et al. Control of T cell activation by CD4+CD25+T suppressor T cells[J]. ImmunolRev, 2001, 182:58-67.
28. Cobbokl SP, Graca L, Lin CY, et al. Regulatory T cells in the induction and maintenance of peripheral transplantation tolerance[J]. Transpl Int, 2003, 16(2) : 66-75.
29. Zorn E, Kim HT, Lee SJ, et al. Reduced frequency of FOXP3+CD4+CD25+regulatory T cells in patients with chronic graft-versus-host disease[J]. Blood, 2005, 106(8) : 2903-2911.
30. Hori S, Nomura T, Sakaguchi S. Control of regulatory T cell development by the transcription factor FoxP3[J]. Science, 2003, 299(5609):1057-1061.
  1. 1. Shapiro AM, Lakey JR, Ryan EA, et al. Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen[J]. N Engl J Med, 2000, 343(4):230-238.
  2. 2. Ryan EA, Paty BW, Senior PA, et al. Five-year follow-up after clinical islet transplantation[J]. Diabetes, 2005, 54(7):2060-2069.
  3. 3. Larsen JL, Bennett RG, Burkman T, et al. Tacrolimus and sirolimus cause insulin resistance in normal sprague dawley rats[J]. Transplantation, 2006, 82(4): 466-470.
  4. 4. Wang J, Xu R, Jin R, et al. Immunosuppressive activity of the Chinese medicinal plant Tripterygium wilfordii. Ⅰ. Prolongation of rat cardiac and renal allograft survival by the PG27 extract and immunosuppressive synergy in combination therapy with cyclosporine[J]. Transplantation, 2000, 70(3): 447-455.
  5. 5. Li R, Takazawa K, Suzuki H, et al. Synergistic effect of triptolideand tacrolimus on rat cardiac allotransplantation[J]. Jpn Heart J, 2004, 45(4):657-665.
  6. 6. Liu Y, Chen Y, Liu FQ, et al. Combined treatment with triptolideand rapamycin prolongs graft survival in a mouse model of cardiactransplantation[J]. Transpl Int, 2008, 21(5):483-494.
  7. 7. Zhang G, Liu Y, Guo H, et al. Triptolide promotes generation of FoxP3+ T regulatory cells in rats[J]. J Ethnopharmacol, 2009, 125(1): 41-46.
  8. 8. 鄭樹森, 郭華, 俞軍, 等. 雷公藤多甙抑制大鼠異種肝移植急性排斥的研究[J]. 中華器官移植雜志, 2003, 24 (6):223-225.
  9. 9. 劉爽, 崔士華, 孫海晨, 等. 一種改良的小鼠胰島分離和移植模型[J]. 首都醫(yī)科大學(xué)學(xué)報(bào), 2009, 30(2):259-261.
  10. 10. Xin MJ, Cui SH, Liu S, et al. Triptolide prolonged allogeneic islet graft survival in chemically induced and spontaneously diabeticmice without impairment of islet function[J]. Hepatobiliary Pancreat Dis Int, 2010, 9(3):312-318.
  11. 11. Liang K, Du W, Zhu W, et al. Contribution of different mecha-nisms to pancreatic beta-cell hyper-secretion in non-obese diabetic(NOD) mice during pre-diabetes[J]. J Biol Chem, 2011, 286(45):39537-39545.
  12. 12. 徐澤寬, 苗毅. 胰腺移植和胰島移植的現(xiàn)狀[J]. 中國普外基礎(chǔ)與臨床雜志, 2003, 10(6): 532-534.
  13. 13. 張劍, 陳規(guī)劃, 翁建平, 等. 抗CD40L 單克隆抗體在異種胰島移植排斥反應(yīng)中的作用[J]. 中國普外基礎(chǔ)與臨床雜志, 2008, 15(5): 329-332.
  14. 14. 李柏峰, 劉永鋒, 程穎, 等. 氨基胍對大鼠胰島保護(hù)作用的實(shí)驗(yàn)研究[J]. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(9): 729-733.
  15. 15. Lo DJ, Anderson DJ, Weaver TA, et al. Belatacept and sirolimusprolong nonhuman primate renal allograft survival without arequirement for memory T cell depletion[J]. Am J Transplant, 2013, 13(2): 320-328.
  16. 16. Lowe M, Badell IR, Thompson P, et al. A novel monoclonal antibody to CD40 prolongs islet allograft survival[J]. Am J Transplant, 2012, 12(8): 2079-2087.
  17. 17. Yin N, Zhang N, Xu J, et al. Targeting lymphangiogenesis after islet transplantation prolongs islet allograft survival[J]. Transplantation, 2011, 92(1) :25-30.
  18. 18. Nataraju A, Saini D, Ramachandran S, et al. Oleanolic acid, a plant triterpenoid, significantly improves survival and function of islet allograft[J]. Transplantation, 2009, 88(8):987-994.
  19. 19. Zhou R, Zhang F, He PL, et al. (5R)-5-hydroxytriptolide (LLDT-8), a novel triptolide analog mediates immunosuppressive effects in vitro and in vivo[J]. Int Immunopharmacol, 2005, 5(13-14):1895-1903.
  20. 20. Liu Q, Chen T, Chen G, et al. Triptolide impairs dendritic cell migration by inhibiting CCR7 and COX-2 expression through PI3-K/Akt and NF-kappaB pathways[J]. Mol Immunol, 2007,44(10): 2686-2696.
  21. 21. Hong Y, Zhou W, Li K, et al. Triptolide is a potent suppressant of C3, CD40 and B7h expression in activated human proximaltubular epithelial cells[J]. Kidney Int, 2002, 62(4):1291-1300.
  22. 22. Liu Y, Chen Y, Lamb JR, et al. Triptolide, a component of Chinese herbal medicine, modulates the functional phenotype of dendritic cells[J]. Transplantation, 2007, 84(11): 1517-1526.
  23. 23. Hoffmann P, Ermann J, Edinger M, et al. Donor-type CD4+CD25+T regulatory T cells suppress lethal acute graft-versus-host disease after allogeneic bone marrow transplantation[J]. J Exp Med, 2002, 196(3): 389-399.
  24. 24. Nguyen VH , Shashidhar S, Chang DS, et al. The impact of regulatory T cells on T-cell immunity following hematopoietic cell transplantation[J] . Blood, 2008, 111(2): 945- 953.
  25. 25. Jiang X, Sun W, Guo D, et al. Cardiac allograft acceptance induced by blockade of CD40-CD40L costimulation is dependent on CD4+CD25+regulatory T cells[J] . Surgery, 2011, 149(3):336-346.
  26. 26. Sakaguchi S. Naturally arising CD4+ regulatory T cells for immu-nologic self tolerance and negative control of immune responses[J]. Annu Rev Immunol, 2004, 22: 531-562.
  27. 27. Shevach EM, McHugh RS, Piccirillo CA, et al. Control of T cell activation by CD4+CD25+T suppressor T cells[J]. ImmunolRev, 2001, 182:58-67.
  28. 28. Cobbokl SP, Graca L, Lin CY, et al. Regulatory T cells in the induction and maintenance of peripheral transplantation tolerance[J]. Transpl Int, 2003, 16(2) : 66-75.
  29. 29. Zorn E, Kim HT, Lee SJ, et al. Reduced frequency of FOXP3+CD4+CD25+regulatory T cells in patients with chronic graft-versus-host disease[J]. Blood, 2005, 106(8) : 2903-2911.
  30. 30. Hori S, Nomura T, Sakaguchi S. Control of regulatory T cell development by the transcription factor FoxP3[J]. Science, 2003, 299(5609):1057-1061.