Osaragi Tomohiko, Nagahiro Itaru, Miyaguchi Naoyuki, Mori Hideaki, Sano Yoshifumi, Date Hiroshi, Shimizu Nobuyoshi
Department of Cancer and Thoracic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan.
Acta Med Okayama. 2008 Oct;62(5):297-302. doi: 10.18926/AMO/30969.
The aim of this study was to determine the optimal temperature of graft preservation after ex vivo gene transfer to rat lung isografts. Left lungs were harvested and infused with cationic lipid/LacZ-DNA complex via the pulmonary artery, and the grafts were stored for 4h. The grafts (n7) were allocated into groups I-IV according to the storage temperature:4, 10, 16, and 23, respectively. Forty-eight h after orthotopic transplantation, the arterial blood gas was analyzed and the peak airway pressure (PAP) and the level of LacZ protein production in the grafts were measured by reverse transcription polymerase chain reaction. After reperfusion, the grafts were stained with hematoxylin and eosin. The grafts in groups III and IV showed more deterioration as evidenced by decreased arterial oxygen tension, increased PAP, and predominant infiltration of inflammatory cells compared with groups I and II. The level of LacZ production was significantly lower in group I than in groups II-IV. The optimal temperature of lung graft preservation after ex vivo gene transfer was determined to be 10, balancing considerations of lung injury and efficiency of transgene expression.
本研究的目的是确定大鼠肺同种异体移植体外基因转移后移植物保存的最佳温度。切取左肺并通过肺动脉注入阳离子脂质/ LacZ - DNA复合物,然后将移植物保存4小时。将移植物(n = 7)根据保存温度分为I - IV组:分别为4℃、10℃、16℃和23℃。原位移植48小时后,分析动脉血气,并通过逆转录聚合酶链反应测量移植物中的气道峰值压力(PAP)和LacZ蛋白产生水平。再灌注后,将移植物用苏木精和伊红染色。与I组和II组相比,III组和IV组的移植物显示出更多的恶化,表现为动脉氧分压降低、PAP升高以及炎症细胞的大量浸润。I组中LacZ产生水平明显低于II - IV组。综合考虑肺损伤和转基因表达效率,确定体外基因转移后肺移植物保存的最佳温度为10℃。