大鼠供肾转染Fas配体基因延长移植肾存活时间

目的观察大鼠供肾转染Fas配体(FasL)基因是否能延长肾移植后受者和移植肾的存活时间,研究其对移植肾的保护作用。方法供者为SD大鼠,受者为Wistar大鼠,随机配对分为实验组和对照组。实验组供肾移植前用1ml含重组腺病毒AdV-FasL的HC-A肾脏冷保存液(0~4℃)经肾动脉灌注;对照组用1mlHC-A肾脏冷保存液(0~4℃)灌注。建立同种肾移植模型。应用逆转录聚合酶链法(RT-PCR)及免疫组织化学方法检测外源FasL基因的表达;透射电镜观察移植肾超微结构的变化;并对肾移植后大鼠的存活率及血肌酐水平进行观察。结果实验组移植肾转基因后FasLmRNA及蛋白均呈阳性表达,FasL蛋白的表达主要分布于小动脉、肾小球及近曲小管。实验组移植肾免疫排斥反应及超微结构变化均较对照组减轻。实验组及对照组肾移植后大鼠的存活时间中位数分别为32.2d和12.1d;术后第7d,实验组血肌酐水平为(315.2±19.2)μmol/L,对照组为(416.2±48.6)μmol/L;两组相比,差异有统计学意义(P<0.05)。结论腺病毒载体可成功介导FasL基因对大鼠肾脏的转移,并对移植肾起免疫保护作用、延长移植肾的存活时间。

The effect of fas ligand gene transfer to renal allografts by adenovirus vector in rats

Objective To investigate the protective effect of Fas ligand (FasL) gene transfer to renal allografts in rats. Methods FasL recombinant adenovirus vector was contruccted and transduced into rat renal allografts by renal artery perfusion. RT-PCR and immunohistochemistry were use d to detect the expression of exogenous FasL gene. Electronic microscopy was use d to observe the changes in the ultra-structure. At the same time, mean surviva l of animals and the level of serum creatinine were observed. Results FasL mRNA and protein were positively expressed in FasL gene transduced renal al lografts. FasL protein mainly distributed in renal small artery, glomerular and proximal tubues. FasL transduced allografts had less renal graft rejection and c hange in the ultra-structure, and prolonged allograft survival and lower serum creatinine as compared to control animals (P< 0.05 ). Conclusions Adenoviral vector can successfully transduce rat kidneys with the FasL cDNA. Fas L gene transfer can protect rat renal allografts from immunologic attack and pro long the allograft survival.