Rentsch Markus, Kienle Klaus, Mueller Thomas, Vogel Mandy, Jauch Karl Walter, Püllmann Kerstin, Obed Aiman, Schlitt Hans J, Beham Alexander
Department of Surgery, Klinikum Grosshadern, Ludwig-Maximilians-University, Munich, Germany.
Transplantation. 2005 Nov 27;80(10):1461-7. doi: 10.1097/01.tp.0000178293.65311.8b.
Primary graft dysfunction due to ischemia and reperfusion injury represents a major problem in liver transplantation. The related cell stress may induce apoptosis, which can be suppressed by bcl-2. The purpose of the study was to investigate the effect of adenoviral bcl-2 gene transfer on early graft function and survival in rat liver transplantation.
An adenoviral construct that transfers bcl-2 under the control of a tetracycline inducible promoter was generated (advTetOn bcl-2) and used with a second adenovirus that transfers the repressor protein (advCMV Rep). Forty-eight hours before explantation, donor rats were treated with advTetOn bcl-2/ advCMV Rep (n=7) and doxycyclin, with the control adenoviral construct advCMV GFP (n=8) or with doxycyclin alone (n=8). Liver transplantation was performed following 16 hours of cold storage (UW). Bcl-2 expression and intrahepatic apoptosis was assessed. Bile flow was monitored 90 min posttransplantation. The endpoint for survival was 7 days.
Bcl-2 was expressed in hepatocytes and sinusoidal lining cells. This was associated with a significant reduction of apoptotic sinusoidal lining cells and hepatocytes after 24 hours and 7 days. Bile production was significantly higher following bcl-2 pretreatment. Furthermore, bcl-2 transfer resulted in significantly improved survival (100% vs. 50% both control groups).
Adenoviral bcl-2 transfer results in protein expression in hepatocytes and sinusoidal lining cells resulting in early graft function and survival enhancement after prolonged ischemia and reperfusion injury. The inhibition of apoptosis in the context of liver transplantation might be a reasonable approach in the treatment of graft dysfunction.
缺血再灌注损伤所致的原发性移植肝无功能是肝移植中的一个主要问题。相关的细胞应激可能诱导细胞凋亡，而bcl-2可抑制这种凋亡。本研究旨在探讨腺病毒介导的bcl-2基因转移对大鼠肝移植早期移植肝功能及存活的影响。
构建一种在四环素诱导型启动子控制下转移bcl-2的腺病毒载体（advTetOn bcl-2），并与另一种转移阻遏蛋白的腺病毒（advCMV Rep）联合使用。在供肝切取前48小时，给供体大鼠注射advTetOn bcl-2/advCMV Rep（n=7）和强力霉素，或注射对照腺病毒载体advCMV GFP（n=8），或仅注射强力霉素（n=8）。在冷保存（UW液）16小时后进行肝移植。评估bcl-2表达及肝内细胞凋亡情况。移植后90分钟监测胆汁流量。存活终点为7天。
bcl-2在肝细胞和肝血窦内皮细胞中表达。这与24小时及7天后凋亡的肝血窦内皮细胞和肝细胞显著减少有关。bcl-2预处理后胆汁分泌明显增多。此外，bcl-2基因转移使存活率显著提高（两个对照组的存活率分别为50%，而bcl-2基因转移组为100%）。
腺病毒介导的bcl-2基因转移导致肝细胞和肝血窦内皮细胞中蛋白表达，从而在长时间缺血再灌注损伤后增强移植肝早期功能并提高存活率。在肝移植中抑制细胞凋亡可能是治疗移植肝无功能的一种合理方法。
