留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

EPO预处理骨髓间充质干细胞预防大鼠肾移植术后急性排斥反应的初步研究

张亚 周松 刘永光 范礼佩 李民 郭颖 赵明

张亚, 周松, 刘永光, 等. EPO预处理骨髓间充质干细胞预防大鼠肾移植术后急性排斥反应的初步研究[J]. 器官移植, 2018, 9(4): 283-289. doi: 10.3969/j.issn.1674-7445.2018.04.008
引用本文: 张亚, 周松, 刘永光, 等. EPO预处理骨髓间充质干细胞预防大鼠肾移植术后急性排斥反应的初步研究[J]. 器官移植, 2018, 9(4): 283-289. doi: 10.3969/j.issn.1674-7445.2018.04.008
Zhang Ya, Zhou Song, Liu Yongguang, et al. Preliminary study of the effect of EPO pretreatment of bone marrow mesenchymal stem cells on preventing acute rejection after renal transplantation in rats[J]. ORGAN TRANSPLANTATION, 2018, 9(4): 283-289. doi: 10.3969/j.issn.1674-7445.2018.04.008
Citation: Zhang Ya, Zhou Song, Liu Yongguang, et al. Preliminary study of the effect of EPO pretreatment of bone marrow mesenchymal stem cells on preventing acute rejection after renal transplantation in rats[J]. ORGAN TRANSPLANTATION, 2018, 9(4): 283-289. doi: 10.3969/j.issn.1674-7445.2018.04.008

EPO预处理骨髓间充质干细胞预防大鼠肾移植术后急性排斥反应的初步研究

doi: 10.3969/j.issn.1674-7445.2018.04.008
基金项目: 

广东省重大科技专项 2015B020226005

广东省省级科技计划项目 2016A020220014

详细信息
    作者简介:

    张亚,男,1990年生,硕士研究生,Email:596187494@qq.com

    通讯作者:

    赵明,男,1962年生,博士研究生导师,主任医师,研究方向为移植免疫,Email:zhaoming02@hotmail.com

  • 中图分类号: R617

Preliminary study of the effect of EPO pretreatment of bone marrow mesenchymal stem cells on preventing acute rejection after renal transplantation in rats

More Information
  • 摘要:   目的  探讨促红细胞生成素(EPO)预处理骨髓间充质干细胞(BMSC)用于预防大鼠肾移植术后急性排斥反应的作用。  方法  将BMSC分为5组,分别为对照组(不含EPO)、A组(EPO终浓度10 IU/mL)、B组(EPO终浓度100 IU/mL)、C组(EPO终浓度500 IU/mL)和D组(EPO终浓度1 000 IU/mL),每组细胞培养24 h和48 h,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法测定BMSC的细胞增殖率。将BMSC分为2组,分别为BMSC组(不含EPO)和EPO-BMSC组(EPO终浓度500 IU/mL),每组细胞培养48 h后,采用蛋白印迹(Western blot)法检测各组BMSC表面趋化因子受体(CXCR)4的蛋白表达水平。以Wistar大鼠为供体,SD大鼠为受体,建立肾移植急性排斥反应模型后,将受体鼠随机分为4组,每组6只,分别为对照组(未进行任何干预)、EPO组(术后立即经尾静脉注射1 mL含有500 IU的EPO溶液)、BMSC组(术后立即经尾静脉注射1 mL的1×106/mL BMSC细胞液)、EPO-BMSC组(经尾静脉注射1 mL500 IU/mL EPO体外培养的含有1×106/mL BMSC细胞液),采用血清肌酐(Scr)试剂盒测定血标本中Scr水平,采用Western blot法检测移植肾组织中干扰素(IFN)-γ和白细胞介素(IL)-4的蛋白表达水平。  结果  培养24 h,各组BMSC增殖率比较,差异均无统计学意义(均为P > 0.05)。培养48 h,与对照组比较,C组(EPO终浓度500 IU/mL)的BMSC增殖率较高,差异有统计学意义(P < 0.05)。与BMSC组比较,EPO-BMSC组BMSC表面CXCR4的蛋白表达水平更高(P < 0.05)。肾移植术后1 d,各组受体鼠的Scr水平比较,差异均无统计学意义(均为P > 0.05);术后5 d,与对照组比较,EPO组、BMSC组和EPO-BMSC组受体鼠的Scr水平均明显降低,差异均有统计学意义(均为P < 0.05),且EPO-BMSC组受体鼠的Scr水平低于EPO组、BMSC组,差异均有统计学意义(均为P < 0.05)。术后1 d和术后5 d,各组受体鼠移植肾组织中IL-4的蛋白表达水平比较,差异均无统计学意义(均为P > 0.05)。术后1 d,与对照组比较,EPO组、BMSC组及EPO-BMSC组受体鼠移植肾组织中IFN-γ蛋白表达水平和IFN-γ/IL-4均有不同程度降低,差异均有统计学意义(均为P < 0.05)。术后5 d,与对照组比较,EPO组、BMSC组和EPO-BMSC组中IFN-γ蛋白表达水平和IFN-γ/IL-4均有不同程度降低,差异均有统计学意义(均为P < 0.05),其中EPO-BMSC组中IFN-γ蛋白表达水平和IFN-γ/IL-4均低于EPO组和BMSC组,差异均有统计学意义(均为P < 0.05)。  结论  EPO预处理BMSC可预防肾移植术后急性排斥反应,保护移植肾功能。

     

  • 图  1  大鼠肾移植过程图片

    A图为供肾的切取和灌注;B图为传统端端吻合法将供肾动静脉缝合到受体肾动静脉;C图为准备开放血流;D图为开放血流后

    Figure  1.  The pictures of renal transplantation process in rats

    图  2  各组BMSC培养24 h和48 h增殖率的比较

    A图为培养24 h;B图为培养48 h,与对照组比较,aP < 0.05

    Figure  2.  Comparison of proliferation rate of BMSC cultured for 24 h and 48 h among each group

    图  3  EPO-BMSC组和BMSC组中BMSC表面CXCR4蛋白表达水平的比较

    与对照组比较,aP < 0.05

    Figure  3.  Comparison of the protein expression level of CXCR4 on the surface of BMSC betweenEPO-BMSC group and BMSC group

    图  4  各组受体鼠移植肾组织中相关细胞因子的蛋白表达

    A图为IFN-γ的蛋白表达水平;B图为IL-4的蛋白表达水平;C图为各组IFN-γ和IL-4的蛋白表达水平及IFN-γ/IL-4的比较,与对照组比较,aP < 0.05,与EPO组比较,bP < 0.05,与BMSC组比较,cP < 0.05

    Figure  4.  Protein expression of cytokines in renal allograft tissues of recipient rats in each group

  • [1] DAZZI F, KRAMPERA M. Mesenchymal stem cells and autoimmune diseases[J]. Best Pract Res Clin Haematol, 2011, 24(1): 49-57. DOI: 10.1016/j.beha.2011.01.002.
    [2] CONTRERAS-KALLENS P, TERRAZA C, OYARCE K, et al. Mesenchymal stem cells and their immunosuppressive role in transplantation tolerance[J]. Ann N Y Acad Sci, 2018, 1417(1): 35-56. DOI: 10.1111/nyas.13364.
    [3] CHAMBERLAIN G, FOX J, ASHTON B, et al. Concise review: mesenchymal stem cells: their phenotype, differentiation capacity, immunological features, and potential for homing[J]. Stem Cells, 2007, 25(11): 2739-2749.
    [4] LOEBINGER MR, JANES SM. Stem cells as vectors for antitumour therapy[J]. Thorax, 2010, 65(4): 362-369. DOI: 10.1136/thx.2009.128025.
    [5] HASHEMI SM, GHODS S, KOLODGIE FD, et al. A placebo controlled, dose-ranging, safety study of allogenic mesenchymal stem cells injected by endomyocardial delivery after an acute myocardial infarction[J]. Eur Heart J, 2008, 29(2): 251-259.
    [6] IKEGUCHI R, KAKINOKI R, OHTA S, et al. Recipient bone marrow-derived stromal cells prolong graft survival in a rat hind limb allotransplantation model[J]. Microsurgery, 2017, 37(6): 632-640. DOI: 10.1002/micr.30128.
    [7] PURROY C, FAIRCHILD RL, TANAKA T, et al. Erythropoietin receptor-mediated molecular crosstalk promotes T cell immunoregulation and transplant survival[J]. J Am Soc Nephrol, 2017, 28(8): 2377-2392. DOI: 10.1681/ASN.2016101100.
    [8] GALLATIN WM, WEISSMAN IL, BUTCHER EC. A cell-surface molecule involved in organ-specific homing of lymphocytes. 1983[J]. J Immunol, 2006, 177(1): 5-9.
    [9] SHEN L, GAO Y, QIAN J, et al. A novel mechanism for endothelial progenitor cells homing: the SDF-1/CXCR4-Rac pathway may regulate endothelial progenitor cells homing through cellular polarization[J]. Med Hypotheses, 2011, 76(2): 256-258. DOI: 10.1016/j.mehy.2010.10.014.
    [10] GHADGE SK, MÜHLSTEDT S, OZCELIK C, et al. SDF-1α as a therapeutic stem cell homing factor in myocardial infarction[J]. Pharmacol Ther, 2011, 129(1): 97-108. DOI: 10.1016/j.pharmthera.2010.09.011.
    [11] TARI K, ATASHI A, KAVIANI S, et al. Erythropoietin induces production of hepatocyte growth factor from bone marrow mesenchymal stem cells in vitro[J]. Biologicals, 2017, 45: 15-19. DOI: 10.1016/j.biologicals.2016.10.010.
    [12] LIU N, TIAN J, CHENG J, et al. Effect of erythropoietin on the migration of bone marrow-derived mesenchymal stem cells to the acute kidney injury microenvironment[J]. Exp Cell Res, 2013, 319(13): 2019-2027. DOI: 10.1016/j.yexcr.2013.04.008.
    [13] LIAO W, ZENG F, KANG K, et al. Lipoxin A4 attenuates acute rejection via shifting TH1/TH2 cytokine balance in rat liver transplantation[J]. Transplant Proc, 2013, 45(6): 2451-2414. DOI: 10.1016/j.transproceed.2013.01.069.
    [14] LI B, TIAN L, DIAO Y, et al. Exogenous IL-10 induces corneal transplantation immune tolerance by a mechanism associated with the altered Th1/Th2 cytokine ratio and the increased expression of TGF-β[J]. Mol Med Rep, 2014, 9(6):2245-2250. DOI: 10.3892/mmr.2014.2073.
    [15] MOTA AP, VILAÇA SS, DAS MERCÊS FL JR, et al. Cytokines signatures in short and long-term stable renal transplanted patients[J]. Cytokine, 2013, 62(2): 302-309. DOI: 10.1016/j.cyto.2013.03.001.
    [16] 金潇, 周松, 胡建敏, 等.间充质干细胞在肾脏移植中的应用研究进展[J].实用医学杂志, 2018, 34(3): 499-502. DOI: 10.3969/j.issn.1006-5725.2018.03.040.

    JIN X, ZHOU S, HU JM, et al. Advances in the application of mesenchymal stem cells in renal transplantation[J]. J Pract Med, 2018, 34(3): 499-502. DOI: 10.3969/j.issn.1006-5725.2018.03.040.
    [17] XIE J, WANG Y, BAO J, et al. Immune tolerance induced by RelB short-hairpin RNA interference dendritic cells in liver transplantation[J]. J Surg Res, 2013, 180(1): 169-175. DOI: 10.1016/j.jss.2012.10.021.
  • 加载中
图(4)
计量
  • 文章访问数:  71
  • HTML全文浏览量:  44
  • PDF下载量:  4
  • 被引次数: 0
出版历程
  • 收稿日期:  2018-04-18
  • 网络出版日期:  2021-01-19
  • 刊出日期:  2018-07-15

目录

    /

    返回文章
    返回