Ricciardi R, Kim R D, McDade T P, Perugini R A, Veal T M, Quarfordt S H, Callery M P, Chari R S, Meyers W C
Department of Surgery, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA.
J Surg Res. 2000 Sep;93(1):35-40. doi: 10.1006/jsre.2000.5961.
In liver transplantation, activation of NFkappaB occurs upon reperfusion, yet few data exist regarding NFkappaB activation during cold ischemia. We hypothesized that activation of NFkappaB may initially occur during cold ischemia, prior to reperfusion, and serve as an important determinant of postreperfusion function. To test this hypothesis, serial biopsies during porcine liver harvest were obtained immediately upon laparotomy, upon completion of dissection, after 45 and 120 min of cold ischemia, and 60 and 180 min after reperfusion. Nuclear extracts were isolated for Western blot analysis of NFkappaB. Hepatic function was assessed through bile output and sorbitol dehydrogenase (SDH) activity. NFkappaB expression was maximal at 45 min of cold ischemia and decreased by 120 min. The expression at 120 min of cold ischemia correlated with markers of postreperfusion function, namely bile flow and SDH activity. During reperfusion a second distinct peak occurred at 180 min. Increased expression of NFkappaB at 180 min of reperfusion correlated directly with prior expression at 120 min during cold ischemia and with increased SDH activity. These data indicate that nuclear expression of NFkappaB demonstrate two distinct peaks of activity, one during cold ischemia and one after reperfusion. Enhanced expression of NFkappaB during cold ischemia not only correlates directly with NFkappaB expression during reperfusion, but also correlates inversely with postreperfusion graft function.
在肝移植中，核因子κB（NFκB）在再灌注时被激活，但关于冷缺血期间NFκB激活的数据却很少。我们推测，NFκB的激活可能最初发生在冷缺血期间，即在再灌注之前，并作为再灌注后功能的一个重要决定因素。为了验证这一假设，在猪肝脏获取过程中，于剖腹术后立即、解剖完成后、冷缺血45分钟和120分钟后以及再灌注60分钟和180分钟后获取系列活检样本。分离核提取物用于NFκB的蛋白质印迹分析。通过胆汁分泌量和山梨醇脱氢酶（SDH）活性评估肝功能。NFκB表达在冷缺血45分钟时达到最大值，并在120分钟时下降。冷缺血120分钟时的表达与再灌注后功能的标志物，即胆汁流量和SDH活性相关。在再灌注期间，在180分钟出现了第二个明显的峰值。再灌注180分钟时NFκB表达的增加与冷缺血期间120分钟时的先前表达直接相关，并且与SDH活性增加相关。这些数据表明，NFκB的核表达呈现出两个不同的活性峰值，一个在冷缺血期间，一个在再灌注后。冷缺血期间NFκB表达的增强不仅与再灌注期间的NFκB表达直接相关，而且与再灌注后移植物功能呈负相关。
