Defamie Virginie, Laurens Marina, Patrono Damiano, Devel Laurent, Brault Antoine, Saint-Paul Marie-Christine, Yiotakis Athanasios, Barbry Pascal, Gugenheim Jean, Crenesse Dominique, Dive Vincent, Huet Pierre-Michel, Mari Bernard
Institut de Pharmacologie Moléculaire et Cellulaire, CNRS, UMR6097, Université de Nice-Sophia Antipolis, France.
Hepatology. 2008 Jan;47(1):177-85. doi: 10.1002/hep.21929.
Matrix metalloproteinases (MMPs) have been implicated in the hepatic injury induced after cold ischemia-warm reperfusion (CI-WR), by altering the extracellular matrix (ECM), but their precise role remains unknown. The hepatic MMP expression was evaluated after 2 conditions of CI (4 degrees C for 24 and 42 hours: viable and nonviable livers) followed by different periods of WR, using isolated perfused rat livers. CI-WR induced moderate changes in hepatic MMP transcript levels not influenced by CI duration, whereas gelatinase activities accumulated in liver effluents. Therefore, the protective effect of a new phosphinic MMP inhibitor, RXP409, was tested after prolonged CI. RXP409 (10 microM) was added to the University of Wisconsin solution, and livers were preserved for 42 hours (4 degrees C), then reperfused for 1 hour in Krebs solution (37 degrees C), containing 20% erythrocytes. Liver viability parameters were recorded, and the extent of cell necrosis was evaluated on liver biopsies, using trypan blue nuclear uptake. Treatment with RXP409 significantly improved liver function (transaminase release and bile secretion) and liver injury. In particular, the MMP inhibitor significantly modified the extent of cell death from large clusters of necrotic hepatocytes as found in control livers (2%-60% of liver biopsies; mean, 26% +/- 9%) to isolated necrotic hepatocytes as found in treated livers (0.2%-12%; mean, 3% +/- 2%) (P < 0.05).
These data demonstrate that MMPs, by altering the ECM, play a major role in liver CI-WR injury leading to extensive hepatocyte necrosis and that their inhibition might prove to be a new strategy in improving preservation solutions.
基质金属蛋白酶（MMPs）通过改变细胞外基质（ECM）参与冷缺血-热再灌注（CI-WR）后诱导的肝损伤，但其确切作用尚不清楚。使用离体灌注大鼠肝脏，在两种CI条件（4℃下24小时和42小时：存活和非存活肝脏）后，再进行不同时间段的WR，评估肝脏MMP表达。CI-WR诱导肝脏MMP转录水平发生中度变化，不受CI持续时间影响，而明胶酶活性在肝脏流出物中积累。因此，在长时间CI后测试了一种新型次膦酸MMP抑制剂RXP409的保护作用。将RXP409（10 microM）添加到威斯康星大学溶液中，肝脏在4℃下保存42小时，然后在含有20%红细胞的克雷布斯溶液（37℃）中再灌注1小时。记录肝脏活力参数，并使用台盼蓝核摄取评估肝活检中细胞坏死程度。用RXP409治疗可显著改善肝功能（转氨酶释放和胆汁分泌）和肝损伤。特别是，MMP抑制剂显著改变了细胞死亡程度，从对照肝脏中发现的大片坏死肝细胞群（肝活检的2%-60%；平均，26%±9%）变为治疗肝脏中发现的孤立坏死肝细胞（0.2%-12%；平均，3%±2%）（P<0.05）。
这些数据表明，MMPs通过改变ECM在导致广泛肝细胞坏死的肝脏CI-WR损伤中起主要作用，抑制它们可能是改善保存溶液的一种新策略。
