Crenesse Dominique, Laurens Marina, Heurteaux Catherine, Cursio Raffaele, Saint-Paul Marie Christine, Schmid-Alliana Annie, Gugenheim Jean
Laboratoire de Physiologie, Faculté de Médecine, Université de Nice-Sophia Antipolis, Av. de Valombrose 06107 Nice 2, France.
Eur J Pharmacol. 2003 Jul 25;473(2-3):177-84. doi: 10.1016/s0014-2999(03)01977-0.
The aim of this study was to demonstrate that tacrolimus (FK506) has a hepatoprotective effect by reducing ischemia-reperfusion-induced apoptosis and necrosis, both of which lead to post-surgical liver dysfunction. An ischemia-reperfusion model and primary cultured rat hepatocytes subjected to hypoxic and reoxygenation phases, mimicking the surgical process, were used. c-Jun N-terminal kinase 1/stress-activated protein kinase 1 (JNK1/SAPK1) activation leads to caspase 3 activation, a trigger of apoptosis. The activation status of JNK1/SAPK1 was evaluated by immunoprecipitation or Western-blotting experiments. Apoptosis was assessed by measuring caspase activation and by TUNEL (terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate-biotin nick-end labeling) reaction. Necrosis was assessed histologically. Tacrolimus improved the survival rate of rats subjected to ischemia-reperfusion. After FK506 pretreatment, the liver necrosis rate was reduced, and ischemia-reperfusion-induced JNK1/SAPK1 activation and apoptosis were significantly decreased. In hypoxia-reoxygenation-subjected hepatocytes, tacrolimus reduced JNK1/SAPK1 and caspase 3 activation. In the liver, tacrolimus prevented ischemia-reperfusion-induced apoptosis and necrosis.
本研究的目的是证明他克莫司（FK506）具有肝脏保护作用，可减少缺血再灌注诱导的凋亡和坏死，这两者都会导致术后肝功能障碍。使用了缺血再灌注模型以及经历缺氧和复氧阶段的原代培养大鼠肝细胞，以此模拟手术过程。c-Jun氨基末端激酶1/应激激活蛋白激酶1（JNK1/SAPK1）的激活会导致半胱天冬酶3激活，而半胱天冬酶3是凋亡的触发因素。通过免疫沉淀或蛋白质印迹实验评估JNK1/SAPK1的激活状态。通过测量半胱天冬酶激活以及TUNEL（末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸生物素缺口末端标记）反应来评估凋亡。通过组织学评估坏死情况。他克莫司提高了经历缺血再灌注大鼠的存活率。FK506预处理后，肝脏坏死率降低，且缺血再灌注诱导的JNK1/SAPK1激活和凋亡显著减少。在经历缺氧复氧的肝细胞中，他克莫司降低了JNK1/SAPK1和半胱天冬酶3的激活。在肝脏中，他克莫司预防了缺血再灌注诱导的凋亡和坏死。
