摘要
目的以大鼠肾移植为载体，研究腺病毒Fas配体重组体（Ad-FasL）局部转染供肾对肾移植排斥反应的抑制作用。方法供者为近交系Lewis大鼠，受者为近交系Wistar大鼠，建立大鼠的肾移植模型。实验分为3组，每组供、受者各8只。（1）Ad-FasL组：供者麻醉后，开腹，夹闭肾血管以下的腹主动脉、下腔静脉和肝门血管，经腹主动脉插管先向肾动脉内注入5ml UW液，随后灌注9×10^9PFU／ml Ad-FasL液1 ml，灌注时间为3min，取左肾作为供肾进行移植，取右肾检测Ad-FasL转染效率；（2）CsA组：受者肾移植术日至术后5d采用CsA5mg／kg灌胃；（3）对照组：实验方法与Ad-FasL组相同，不同的是灌注液为AdV-5（Pfgl40）空载体。术后比较各组的存活时间、移植肾组织学、肾功能和免疫组织化学等的改变。结果Ad-FasL组受者平均存活时间为31．3d，其中2例超过48d；CsA组平均存活为16．5d；对照组平均存活为9．1d；Ad-FasL组与CsA组和对照组比较，差异均有统计学意义。Ad-FasL组的移植肾功能基本保持稳定，对照组在术后5d切除对侧肾脏后血清肌酐浓度迅速上升；CsA组血清肌酐浓度呈现缓慢上升趋势。结论Ad-FasL能成功转染大鼠供肾；转染Ad-FasL能使移植肾存活时间延长。Ad-FasL对淋巴细胞增殖具有明显的抑制作用，从而抑制了肾移植排斥反应。
Objective To study adenovirus （Ad）-FasL recombinant inhibiting renal-graft rejection in rats. Methods Allogeneic kidney transplants were performed between Lewis donors and Wistar recipients. The experimental rats were divided into three groups. In Ad-FasL group, the donor kidney of the Lewis rat was locally transfected by Ad-FasL and then transplanted to the recipient Wistar rat. In CsA group, the donor kidney of the Lewis rat was directly transplanted to the recipient Wistar rat, but after the operation, the recipient was treated by CsA. In control group, the donor kidney of the Lewis rat was directly transplanted to the recipient Wistar rat without any therapy. Donor kidneys were perfused in situ with Ad-5 （control）, or 9 109 plaque forming units of Ad-FasL. Kidney allografts harvested at 7th day were evaluated by HE staining and irrtmunohistochemical staining. Transduced allografts were stained for the FasL expression using a monoclonal antibody and tested for FasL mRNA production by reverse transcriptase-polymerase chain reaction. Results Through animal exper- iment, it was found the mean survival time in Ad-FasL, control and CsA group was 31.3, 9. 1 and 16. 5 days respectively days. The mean survival time increased 22 days in Ad-FasL treated group. There was significant difference between Ad-FasL group and control group or CsA group （P〈0. 05）. Conclusions Adenoviral vectors can be successfully transduced into rat kidneys with the FasL cDNA. Ad-mediated transduction of the FasL gene can significantly prolong the survival of rat renal allograft. The results of in vitro studies suggested that FasL-mediated cytotoxicity to the activated Fas-expressing cells contributes, at least in part, to the delayed graft rejection in this model. Ad-FasL can induce immune tolerance of renal transplantation.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2008年第2期78-81,共4页
Chinese Journal of Organ Transplantation
关键词
配体
器官移植
移植物排斥
腺病毒科
Ligands
Organ transplantation
Graft rejection
Adenovirdae
