Takei Y, Marzi I, Kauffman F C, Currin R T, Lemasters J J, Thurman R G
Department of Pharmacology, University of North Carolina, Chapel Hill 27599.
Transplantation. 1990 Jul;50(1):14-20. doi: 10.1097/00007890-199007000-00003.
Kupffer cells are activated by calcium and release a variety of toxic mediators, including proteases. The purpose of these studies, therefore, was to determine if protease inhibitors and a calcium channel blocker could increase survival time in the rat model of orthotopic liver transplantation. Survival for 30 days was greater than 90% in this model when livers were stored for 1 hr in Ringer's solution (survival conditions)--however, grafts stored for 4 hr in Euro-Collins solution or 8 hr in University of Wisconsin (UW) solution survived postoperatively only 1.2 and 0.7 days, respectively (nonsurvival conditions). When livers were stored for 4 hr in Euro-Collins containing a cocktail of protease inhibitors (leupeptin, pepstatin A, phenylmethylsulfonyl fluoride, 20 ng/ml each; diisopropyl fluorophosphate, 100 microM) and subsequently transplanted, however, survival time was increased significantly to 11.5 days. Inclusion of a calcium channel blocker, nisoldipine (1.4 microM), in the protease inhibitor cocktail increased survival time to 23 days. Actually, nisoldipine alone increased survival time to 25 days. Nisoldipine alone also increased survival time in livers stored for 8 or 16 hr in UW solution to between 15 and 20 days. Serum transaminase levels reached peak values greater than 2400 U/L one day postoperatively in the nonsurvival groups, and liver injury assessed histologically was apparent. Under these conditions, pulmonary infiltration of inflammatory cells was observed in about 60% of the lungs examined and was associated with massive bleeding. Inclusion of the protease cocktail, nisoldipine, or both in the storage solutions decreased maximal SGOT levels and injury to both liver and lung significantly by about 50% postoperatively. Nisoldipine also decreased phagocytosis of carbon particles by the perfused liver 2- to 3-fold following storage under nonsurvival conditions (half-maximal effect = 0.3-0.4 microM nisoldipine). Moreover, nisoldipine improved hepatic microcirculation. It accelerated blood flow into the liver, as indexed by hemoglobin reflectance from the liver surface. These data support the hypothesis that Kupffer cells are activated early in the sequence of events that causes graft failure leading to endothelial cell-mediated alterations in the microcirculation. This work demonstrates clearly that dihydropyridine-type calcium channel blockers such as nisoldipine may be clinically useful in storage solutions for liver prior to transplantation.
库普弗细胞被钙离子激活并释放多种毒性介质，包括蛋白酶。因此，这些研究的目的是确定蛋白酶抑制剂和钙通道阻滞剂是否能延长原位肝移植大鼠模型的存活时间。在该模型中，当肝脏在林格氏液中保存1小时（存活条件）时，30天的存活率大于90%——然而，在欧洲柯林斯液中保存4小时或在威斯康星大学（UW）液中保存8小时的移植物术后分别仅存活1.2天和0.7天（非存活条件）。然而，当肝脏在含有蛋白酶抑制剂混合物（亮抑酶肽、胃蛋白酶抑制剂A、苯甲基磺酰氟，各20 ng/ml；二异丙基氟磷酸，100 μM）的欧洲柯林斯液中保存4小时，随后进行移植时，存活时间显著延长至11.5天。在蛋白酶抑制剂混合物中加入钙通道阻滞剂尼索地平（1.4 μM）可使存活时间延长至23天。实际上，单独使用尼索地平可使存活时间延长至25天。单独使用尼索地平也可使在UW液中保存8或16小时的肝脏的存活时间延长至15至20天。非存活组术后1天血清转氨酶水平达到峰值大于2400 U/L，组织学评估的肝损伤明显。在这些条件下，在约60%检查的肺中观察到炎性细胞的肺浸润，并伴有大量出血。在保存液中加入蛋白酶混合物、尼索地平或两者，可使术后最大谷草转氨酶水平以及肝和肺损伤显著降低约50%。在非存活条件下保存后，尼索地平还使灌注肝脏对碳颗粒的吞噬作用降低2至3倍（半数最大效应 = 0.3 - 0.4 μM尼索地平）。此外，尼索地平改善了肝微循环。它加速了血液流入肝脏，以肝脏表面血红蛋白反射率为指标。这些数据支持这样的假说，即库普弗细胞在导致移植物衰竭的一系列事件早期被激活，进而导致内皮细胞介导的微循环改变。这项工作清楚地表明，诸如尼索地平之类的二氢吡啶型钙通道阻滞剂在肝移植前的保存液中可能具有临床应用价值。
