Caspase 3 inhibition improves survival and reduces early graft injury after ischemia and reperfusion in rat liver transplantation
Caspase 3 inhibition improves survival and reduces early graft injury after ischemia and reperfusion in rat liver transplantation
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DOI：
10.1097/01.tp.0000141095.06273.10
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发表时间：
2004-11-15
期刊：
影响因子：
6.2
通讯作者：
Rentsch, M
中科院分区：
文献类型：
--
作者：
Mueller, THJ;Kienle, K;Rentsch, M
背景细胞凋亡在器官移植缺血再灌注后起着重要作用。它由半胱天冬酶执行，并受bcl-2家族的促凋亡蛋白和抗凋亡蛋白的变阻器影响。本研究探讨了特异性抑制caspase 3和7对肝移植后移植物功能、存活率和肝脏bcl-2水平的影响。刘易斯大鼠在冷移植物储存16小时后进行同系原位肝移植（在威斯康星州大学溶液中）。研究了用D（OMe）E（OMe）VD（OMe）-氟甲基酮（细胞凋亡执行器半胱天冬酶3和7的特异性抑制剂）处理的供体动物的肝脏和适当的对照组。通过测量胆汁流量和使用体内荧光显微镜测定微血管移植物损伤来量化早期移植物损伤。通过末端脱氧核苷酸转移酶介导的dUTP缺口末端标记染色和细胞死亡效应物的Western blot分析来检测细胞凋亡及其调控。在体内特异性半胱天冬酶抑制后，蛋白质印迹分析显示半胱天冬酶诱导的聚ADP-核糖聚合酶裂解的抑制。半胱天冬酶3和7的抑制导致凋亡内皮细胞的数量显着减少，并改善微血管灌注。在第7天bcl-2水平的增加也表明了细胞保护作用。最重要的是，特异性半胱天冬酶阻断可提高大鼠肝移植后的存活率。特异性抑制凋亡执行蛋白半胱天冬酶能有效减轻肝移植缺血再灌注损伤，提高移植物存活率。半胱天冬酶抑制后更好的组织保存与减少的凋亡执行，改善微血管灌注和bcl-2上调相关。因此，特异性半胱天冬酶抑制剂代表了临床用于肝移植的有前景的方案。
Background. Apoptosis plays a crucial role after ischemia-reperfusion in organ transplantation. It is executed by caspases and influenced by the rheostat of pro- and anti-apoptotic proteins of the bcl-2 family. This study investigated the effect of specific inhibition of caspases 3 and 7 on graft function, survival, and hepatic bcl-2 levels after liver transplantation.Methods. Lewis rats underwent syngeneic orthotopic liver transplantation after 16 hr of cold graft storage (in University of Wisconsin solution). Livers of donor animals treated with D(OMe)E(OMe)VD(OMe)-fluoromethylketone (specific inhibitor of apoptosis executor caspases 3 and 7), and appropriate control groups, were investigated. Early graft injury was quantified by measurement of bile flow and determination of microvascular graft injury by using in vivo fluorescence microscopy. Apoptosis and its regulation were examined by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling staining and Western blot analysis of cell death effectors, respectively.Results. After specific in vivo caspase inhibition, Western blot analysis revealed inhibition of caspase-induced cleavage of poly-ADP-ribose-polymerase. Inhibition of caspases 3 and 7 resulted in a significantly decreased number of apoptotic endothelial cells and improved microvascular perfusion. A cell protective effect was also suggested by an increase of bcl-2 levels at 7 days. Most important, specific caspase blockade resulted in improved rat survival after liver transplantation.Conclusion. Specific inhibition of apoptosis executor caspases effectively reduces graft ischemia-reperfusion injury and improves survival in liver transplantation. Better tissue preservation after caspase inhibition correlates with reduced apoptosis execution, improved microvascular perfusion, and bcl-2 up-regulation. Therefore, specific caspase inhibition represents a promising regimen for clinical use in liver transplantation.