Yano M, Boasquevisque C H, Scheule R K, Botney M D, Cooper J D, Patterson G A
Division of Cardiothoracic Surgery, Washington University School of Medicine, St. Louis, Mo., USA.
J Thorac Cardiovasc Surg. 1997 Nov;114(5):793-801; discussion 801-2. doi: 10.1016/S0022-5223(97)70083-7.
Gene transfer to lung grafts may be useful in ameliorating ischemia-reperfusion injury and rejection. Efficient gene transfection to the whole organ may prove problematic. Proximal pulmonary artery endothelial transfection might provide beneficial downstream effects on the whole graft. The aim of this study was to determine the feasibility of transfecting proximal pulmonary artery segments in lung isografts.
Male Fischer rats were divided into six groups. In vivo transfection: In group I (n = 7), a proximal segment of the left pulmonary artery was isolated and injected with saline solution by means of a catheter inserted through the right ventricle. After an exposure period of 20 minutes, clamps were removed and blood flow was restored. In group II (n = 7), the isolated arterial segments were injected with adenovirus carrying the Escherichia coli LacZ gene encoding for beta-galactosidase. Ex vivo transfection: In group III (n = 5), arterial segments were injected ex vivo with saline solution and in group IV (n = 5) with the adenovirus construct. In group V (n = 6), arteries were injected with saline solution and in group VI (n = 11) with liposome chloramphenicol acetyl transferase cDNA. In groups I to IV, animals were killed on postoperative day 3 and transgene expression was assessed by Bluo-Gal staining. In groups V and VI, animals were killed on postoperative day 2 and transgene expression was assessed by chloramphenicol acetyl transferase activity assay.
Transgene expression was detected grossly and microscopically in endothelial and smooth muscle cells of pulmonary artery segments from all surviving animals of groups II and IV. In group VI, chloramphenicol acetyl transferase activity was significant in all assessed arterial segments.
Significant transgene expression is observed in proximal pulmonary artery segments after both in vivo and ex vivo exposure.
将基因导入肺移植器官可能有助于减轻缺血再灌注损伤和排斥反应。对整个器官进行高效基因转染可能存在问题。近端肺动脉内皮转染可能会对整个移植器官产生有益的下游效应。本研究的目的是确定在肺同基因移植中对近端肺动脉段进行转染的可行性。
将雄性Fischer大鼠分为六组。体内转染：在第一组（n = 7）中，分离左肺动脉的近端段，通过插入右心室的导管注入盐溶液。暴露20分钟后，松开夹子并恢复血流。在第二组（n = 7）中，向分离的动脉段注入携带编码β-半乳糖苷酶的大肠杆菌LacZ基因的腺病毒。体外转染：在第三组（n = 5）中，将动脉段在体外注入盐溶液，在第四组（n = 5）中注入腺病毒构建体。在第五组（n = 6）中，向动脉注入盐溶液，在第六组（n = 11）中注入脂质体氯霉素乙酰转移酶cDNA。在第一组至第四组中，动物在术后第3天处死，通过Bluo-Gal染色评估转基因表达。在第五组和第六组中，动物在术后第2天处死，通过氯霉素乙酰转移酶活性测定评估转基因表达。
在第二组和第四组所有存活动物的肺动脉段内皮细胞和平滑肌细胞中均在大体和显微镜下检测到转基因表达。在第六组中，所有评估的动脉段中氯霉素乙酰转移酶活性均显著。
在体内和体外暴露后，近端肺动脉段均观察到显著的转基因表达。
