常温灌流和蛋白质补充对器官保存过程中人类内皮细胞功能的影响。

Department of Cardiothoracic Surgery, University Medical Center Regensburg, Regensburg, Germany.

Ann Thorac Surg. 2010 Feb;89(2):512-20. doi: 10.1016/j.athoracsur.2009.10.037.

BACKGROUND

Hypothermia-induced changes in endothelial cell (EC) morphology and function after organ storage may influence the initial outcome and development of transplant-associated coronary artery disease.

METHODS

Human saphenous vein ECs were incubated with saline (NaCl), University of Wisconsin (UW), and histidine-tryptophan-ketoglutarate (HTK) solution, with and without protein additives, at 4 degrees C and 37 degrees C. After 6 hours, ECs were recultivated for 24 and 48 hours with culture medium (reperfusion). Mitochondrial activity, adenosine triphosphate concentration, cell count, and inflammatory responses were analyzed.

RESULTS

Cold preservation did not affect the mitochondrial activity of ECs and allowed a complete regeneration of the metabolic turnover after reperfusion. However, under normothermic conditions the metabolism of the cells was influenced by time and type of preservation solution. While both the mitochondrial activity and cell count did not change after treatment with NaCl and culture medium, the metabolic turnover of cells treated with HTK and UW solution significantly increased (twofold) and decreased (twofold, p < 0.05), respectively, after reperfusion. The endothelial reactivity remained unchanged after treatment with NaCl and HTK. The addition of serum proteins significantly improved mitochondrial activity of cells treated with warm NaCl and HTK (p < 0.05). The UW-treated cells burned out through a significant up-regulation of the ATP concentration resulting in a complete metabolic regression after reperfusion and induction of apoptosis.

CONCLUSIONS

Normothermic preservation in UW prevented regeneration of ECs, while treatment with HKT solution did not irreversibly affect mitochondrial activity of ECs and allowed complete regeneration of metabolism and function. Serum proteins improved the preservation effect of HTK and NaCl.

背景

器官保存过程中低温诱导的内皮细胞（EC）形态和功能变化可能影响移植相关冠状动脉疾病的初始结果和发展。

方法

将人隐静脉 EC 在 4°C 和 37°C 下分别用生理盐水（NaCl）、威斯康星大学（UW）和组氨酸-色氨酸-酮戊二酸（HTK）溶液孵育，有和没有蛋白质添加剂。孵育 6 小时后，将 EC 用培养基（再灌注）在 24 和 48 小时后重新培养。分析线粒体活性、三磷酸腺苷浓度、细胞计数和炎症反应。

结果

低温保存不影响 EC 的线粒体活性，并允许再灌注后代谢周转完全恢复。然而，在常温条件下，细胞的代谢受到保存液类型和时间的影响。虽然在用 NaCl 和培养基处理后，线粒体活性和细胞计数没有变化，但在用 HTK 和 UW 溶液处理后，细胞的代谢周转分别显著增加（两倍）和减少（两倍，p<0.05）。在用 NaCl 和 HTK 处理后，内皮反应性保持不变。添加血清蛋白可显著提高用温热 NaCl 和 HTK 处理的细胞的线粒体活性（p<0.05）。UW 处理的细胞通过显著上调 ATP 浓度而耗尽，导致再灌注后完全代谢衰退和细胞凋亡诱导。