Jawad Rim, D'souza Melroy, Selenius Lisa Arodin, Lundgren Marita Wallenberg, Danielsson Olof, Nowak Greg, Björnstedt Mikael, Isaksson Bengt
Division of Pathology, Department of Laboratory Medicine, Karolinska Institutet, Stockholm S-141 86, Sweden.
Department of Clinical Science, Intervention, and Technology (CLINTEC), Division of Surgery, Karolinska Institutet, Karolinska University Hospital, Huddinge, Stockholm S-141 86, Sweden.
World J Hepatol. 2017 Dec 8;9(34):1261-1269. doi: 10.4254/wjh.v9.i34.1261.
To study the effects of warm ischemia-reperfusion (I/R) injury on hepatic morphology at the ultrastructural level and to analyze the expression of the thioredoxin (TRX) and glutaredoxin (GRX) systems.
Eleven patients undergoing liver resection were subjected to portal triad clamping (PTC). Liver biopsies were collected at three time points; first prior to PTC (baseline), 20 min after PTC (post-ischemia) and 20 min after reperfusion (post-reperfusion). Electron microscopy and morphometry were used to study and quantify ultrastructural changes, respectively. Additionally, gene expression analysis of TRX and GRX isoforms was performed by quantitative PCR. For further validation of redox protein status, immunogold staining was performed for the isoforms GRX1 and TRX1.
Post-ischemia, a significant loss of the liver sinusoidal endothelial cell (LSEC) lining was observed (P = 0.0003) accompanied by a decrease of hepatocyte microvilli in the space of Disse. Hepatocellular morphology was well preserved apart from the appearance of crystalline mitochondrial inclusions in 7 out of 11 patients. Post-reperfusion biopsies had similar features as post-ischemia with the exception of signs of a reactivation of the LSECs. No changes in the expression of redox-regulatory genes could be observed at mRNA level of the isoforms of the TRX family but immunoelectron microscopy indicated a redistribution of TRX1 within the cell.
At the ultrastructural level, the major impact of hepatic warm I/R injury after PTC was borne by the LSECs with detachment and reactivation at ischemia and reperfusion, respectively. Hepatocytes morphology were well preserved. Crystalline inclusions in mitochondria were observed in the hepatocyte after ischemia.
在超微结构水平研究热缺血再灌注（I/R）损伤对肝脏形态的影响，并分析硫氧还蛋白（TRX）和谷氧还蛋白（GRX）系统的表达。
11例行肝切除术的患者接受门静脉三联阻断（PTC）。在三个时间点采集肝活检组织；第一个时间点在PTC前（基线），第二个时间点在PTC后20分钟（缺血后），第三个时间点在再灌注后20分钟（再灌注后）。分别使用电子显微镜和形态计量学来研究和量化超微结构变化。此外，通过定量PCR对TRX和GRX亚型进行基因表达分析。为进一步验证氧化还原蛋白状态，对GRX1和TRX1亚型进行免疫金染色。
缺血后，观察到肝窦内皮细胞（LSEC）衬里显著丧失（P = 0.0003），同时狄氏间隙内肝细胞微绒毛减少。除11例患者中有7例出现结晶状线粒体包涵体外，肝细胞形态保存良好。再灌注后活检组织具有与缺血后相似的特征，但LSECs有重新激活的迹象。在TRX家族亚型的mRNA水平未观察到氧化还原调节基因表达的变化，但免疫电子显微镜显示TRX1在细胞内重新分布。
在超微结构水平，PTC后肝脏热I/R损伤的主要影响由LSECs承担，分别在缺血和再灌注时出现脱离和重新激活。肝细胞形态保存良好。缺血后在肝细胞中观察到线粒体中的结晶状包涵体。
