Knecht Camila, Balaban Cecilia L, Rodríguez Joaquín V, Ceccarelli Eduardo A, Guibert Edgardo E, Rosano Germán L
Instituto de Biología Molecular y Celular de Rosario (IBR), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, 2000, Argentina; Centro Binacional (Argentina-Italia) de Investigaciones en Criobiología Clínica y Aplicada (CAIC), Universidad Nacional de Rosario, Rosario, 2000, Argentina.
Centro Binacional (Argentina-Italia) de Investigaciones en Criobiología Clínica y Aplicada (CAIC), Universidad Nacional de Rosario, Rosario, 2000, Argentina.
Cryobiology. 2018 Dec;85:47-55. doi: 10.1016/j.cryobiol.2018.10.003. Epub 2018 Oct 5.
Cold storage is a common procedure for liver preservation in a transplant setting. However, during cold ischemia, the liver suffers molecular alterations that can affect its performance. Also, deleterious mechanisms set forth in the storage phase are exacerbated during reperfusion. This study aimed to identify liver proteins associated with injury during cold storage and/or normothermic reperfusion using the isolated perfused rat liver model. Livers from male rats were subjected to either (1) cold storage for 24 h, (2) ex vivo normothermic reperfusion for 90 min or (3) cold storage for 24 h followed by ex vivo normothermic reperfusion for 90 min. Then, the livers were homogenized and proteins were extracted. Protein expression between each experimental group and the control (freshly resected livers) was compared by two-dimensional (2D) gel electrophoresis. Protein identification was carried out by matrix-assisted laser desorption/ionization time-of-flight spectrometry (MALDI-TOF/TOF) using MASCOT as the search engine. 23 proteins were detected with significantly altered levels of expression among the different treatments, including molecular chaperones, antioxidant enzymes, and proteins involved in energy metabolism. Some of them have been postulated as biomarkers for liver damage while others had been identified in other organs subjected to ischemia and reperfusion injury. The whole data set will be a useful resource for studying deleterious molecular mechanisms that result in diminished liver function during storage and subsequent reperfusion.
在肝脏移植过程中,冷藏是一种常用的肝脏保存方法。然而,在冷缺血期间,肝脏会发生分子改变,这可能会影响其功能。此外,在再灌注过程中,保存阶段出现的有害机制会加剧。本研究旨在使用离体灌注大鼠肝脏模型,鉴定与冷藏和/或常温再灌注期间损伤相关的肝脏蛋白质。将雄性大鼠的肝脏进行以下处理:(1)冷藏24小时;(2)体外常温再灌注90分钟;(3)冷藏24小时后,再进行体外常温再灌注90分钟。然后,将肝脏匀浆并提取蛋白质。通过二维(2D)凝胶电泳比较每个实验组与对照组(新鲜切除的肝脏)之间的蛋白质表达。使用MASCOT作为搜索引擎,通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF/TOF)进行蛋白质鉴定。在不同处理中检测到23种蛋白质的表达水平有显著变化,包括分子伴侣、抗氧化酶和参与能量代谢的蛋白质。其中一些已被假定为肝损伤的生物标志物,而其他一些则在遭受缺血再灌注损伤的其他器官中被鉴定出来。整个数据集将成为研究导致肝脏在保存和随后再灌注期间功能下降的有害分子机制的有用资源。