›› 2005, Vol. 4 ›› Issue (3): 157-161.

• 基础研究 • 上一篇    下一篇

核因子-κB诱捕物ODN对大鼠移植肾的转染及对细胞因子基因表达的影响

张宏涛 赵显国 丰贵文   

  1. 450000 郑州,郑州大学第一附属医院血液净化中心 
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-03-12 发布日期:2005-03-12

  • Received:1900-01-01 Revised:1900-01-01 Online:2005-03-12 Published:2005-03-12

摘要:

目的 探讨核因子-κB(NFκB)诱补物寡聚核苷酸(ODN)对大鼠供肾的转染效率及对移植肾细 胞因子基因表达影响。方法 建立大鼠移植肾模型,应用含有1μM Liposome/ODN Euro-Collins保存液(Ecs)灌注移植肾并低温缺氧保存6h后移植到受体大鼠;24只大鼠分成4组:假手术组(control组),ECs组,NFκB诱补物(decoy ODN)组,错配ODN(scrambled ODN)组。应用荧光显微镜和多光子共聚焦显微镜观察转染有FITC标记的ODN移植肾,并观看其分布;用RT-PCR技术,检测术后移植肾中ICAM-1、VCAM-1、CINC的mRNA表达。结果 移植术后15min,FITC标记的decoy ODN主要分布在移植肾的肾小球,并且有部分ODN进入细胞核;在术后6h时,诱补物ODN主要分布在肾小球和近曲肾小管中,并且大部分进入细胞核。而不含有脂质体诱补物ODN复合物的Ecs保存的移植肾中则无荧光颗粒;与control组相比,ECs组移植肾,ICAM-1、VCAM-1、CINC 的mRNA水平明显升高 (P<0.05);与 ECs组相比,NF-κB诱捕物ODN处理组移植肾的ICAM-1、VCAM-1、CINC的mRNA水平显著下降( P<0.05),而 Scram. ODN处理组移植肾的ICAM-1、VCAM-1、CINC的mRNA表达则未见抑制( P>0.05)。结论 诱捕物 ODN脂质体复合物在ECs中及低温缺氧条件下可较好地转染大鼠肾脏;降低移植肾的ICAM-1、VCAM-1、CINC的mRNA表达水平。

关键词: NFκB诱补物ODN, 细胞因子, 肾移植

Abstract:

Objective To evaluate the transfection efficiency of nuclear factor κB(NFκB)decoy Oligodeoxynucletides (ODN)into donor kidney and the influnence of decoy ODN on the expression of chemokines of the renal grafts. Methods The rats were divided into 4 groups. Control group: the Wistar rats underwent the sham operation and the kidney was harvested after 6h. ECs roup: The donor kidney was stored for 6h under of hypothermic and hypoxia condition in ECs without ODN/Liposome complexes, then the donor kidney was transplanted into a SD recipient. The allograft was harvested at 6h posttransplantation. Decoy ODN group: The donor kidney was stored for 6h in ECs containing 1μM Liposome/decoy ODN complexes under hypothermic and hypoxia condition, then the donor kidney was transplanted into a SD recipient. The allograft was harvested at 6h posttransplantation. Scrambled ODN group: The donor kidney was stored for 6h in ECs containing 1μM Liposome/scrambled ODN complexes under hypothermal and hypoxia condition, then the donor kidney was transplanted into a SD recipient, and at 6h posttransplantation the allograft was harvested. The distribution of FITC-labeled ODN in renal tissue were observed by fluorescence microscope and TCS SP2 Confocal microscope. The gene expression of ICAM-1 VCAM-1 CINC in renal allografts were detected by RT-PCR. Result FITC-labeled ODN was mainly distributed in glomeruli of the transfected renal allograft at 15min posttransplantation, and mainly in glomeruli and the renal tubules at 6h posttransplantation. The most of ODN was transduced into nuclei at 6h posttransplantation. The fluroscence cann`t be seen in the ECs group. Compared to the control group, the gene expression of ICAM-1, VCAM-1,CINC was observably increased in ECs group ( P>0.05); it was significantly inhibited in the decoy ODN group ( P<0.05) compared to the ECs group and the scram.ODN group. There was no significant difference between the ECs group and the scram.ODN group ( P>0.05). Conclusion Decoy ODN can be efficiently transfected into kidney using in vivo liposome method in ECs under hypothermic and hypoxic condition; The renal allo- graft treated with decoy ODN can inhibit the gene expression of ICAM-1, VCAM-1, CINC.

Key words: Kidney transplantation, Chemokines

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