摘要
目的 探讨CTLA 4Ig基因重组腺病毒 (AdvCTLA 4Ig)有效转染大鼠肾脏的方法并初步观察该基因转染对移植肾存活的影响。方法 AdvCTLA 4Ig对大鼠肾脏分别转染 3h ,包括 :4℃下转染 (A组 )、经肾动脉注射在体转染 (B组 )、阻断肝脏血流以及肾血管以下躯体血流后经肾动脉注射在体转染 (C组 )以及对照 ;以免疫荧光法检测基因转染效率 ;以转染效率最高组肾脏行同种移植 ,未转染组为对照 ,观察移植肾存活时间。结果 A组、B组以及对照组肾脏内未见荧光显示 ,C组肾小管显示阳性荧光 ,肾小球内未见荧光显示 ;各组肾脏光镜下结构正常 ;转染组移植肾存活时间较对照组明显延长 ( 2 6 6± 6 9)dvs ( 8 1± 1 60 )d ,P <0 0 1。结论 高滴度CTLA 4Ig基因重组腺病毒可使在体肾脏中肾小管获得有效转染 ;
Objective To explore the methods of effective transfection of CTLA 4Ig gene into rat kidneys and the effects of gene transfection on the survival time of renal allografts. Methods BN (Brown Norway, RT.1n) rats were randomized into 4 groups according to the methods of kidney transfection by CTLA 4Ig gene recombinant adenovirus (AdvCTLA 4Ig, 5×10 9 pfu/ml): transfection at 4 ℃ in vitro (group A), transfection via renal artery injection of AdvCTLA 4Ig (group B), transfection via renal artery injection of AdvCTLA 4Ig after ligation of the liver vessel and the aorta and the vena cava below the renal vessel (group C) and the control group (D). The transfection efficiency of AdvCTLA 4Ig was determined by immunofluorescence method. The kidneys with the highest transfection efficiency were transplanted into Lewis rats (transfection group). BN→Lewis kidney transplantation with non transplanted kidneys served as the controls. The allograft survival time was observed. Results The results showed that no specific fluorescence was observed in kidneys in group A, group B, and the control group. However, evident specific fluorescence was observed in kidneys in group C. The transfection occurred in tubular cells, but no transfection was observed in glomerular structure. The allograft survival time in transfection group was significantly longer than that of the controls [(26.6±6.9) d vs (8.1±1.6) d, P < 0.01]. Conclusion The rat kidney (mainly the tubular cells) could be transfected efficiently by high titer AdvCTLA 4Ig in vivo , and the survival time of renal allografts could be prolonged by local transfection of CTLA 4Ig gene.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2004年第5期390-392,共3页
Journal of Third Military Medical University
关键词
基因
转染
肾脏移植
gene
transfection
kidney transplantation
