Klöppel K, Gerlach J, Neuhaus P
Chirurgische Klinik und Poliklinik, Universitätsklinikum Rudolf Virchow, Freie Universität Berlin.
Langenbecks Arch Chir. 1994;379(5):264-70. doi: 10.1007/BF00186391.
In order to answer the question of whether there is an optimal buffer system for the preservation and reoxygenation period in liver transplantation, sodium/potassium phosphate, HEPES, TRIS (THAM), MOPS and histidine/His-HCl buffers were investigated. The buffers were added to an "extracellular" electrolyte composition of preservation solution. The solutions were incubated with in vitro cultures of pig hepatocytes in two different models. I: during cold hypoxia (4 degrees C, PO2 < 0.1 mmHg) for 24 h, and II. during the reoxygenation period of 3 h after preservation in UW solution. Cell viability, cell detachment rate, and LDH and GOT liberation were used as parameters of cell alteration. The lowest amount of enzyme release during the preservation period and reoxygenation was obtained using sodium or potassium phosphate buffer. Rising LDH and GOT liberation rates during preservation and reoxygenation were observed with HEPES and TRIS buffer. The enzyme release induced by these three buffer systems correlated with their pKa values. Higher pH of the preservation solution resulted in higher enzyme leakage from the cells. In contrast, the Histidine/HCl buffer system with low pH led to striking cell damage during preservation as well as during reoxygenation. MOPS, a weak acid with the lowest pH in solution, led to the lowest enzyme release during the preservation period, but to high enzyme release after reoxygenation with standard medium. Incubation of the cultures with MOPS after UW preservation resulted in lower enzyme levels in comparison to the controls. In summary, PBS had the best results in our study.
为了回答肝移植保存和复氧期是否存在最佳缓冲系统的问题，对磷酸钠/钾、HEPES、TRIS（三羟甲基氨基甲烷）、MOPS和组氨酸/盐酸组氨酸缓冲液进行了研究。将这些缓冲液添加到保存液的“细胞外”电解质组成中。将这些溶液在两种不同模型中与猪肝细胞的体外培养物一起孵育。I：在冷缺氧（4℃，PO2 < 0.1 mmHg）条件下24小时，以及II：在UW溶液中保存后3小时的复氧期。细胞活力、细胞脱离率以及乳酸脱氢酶（LDH）和谷草转氨酶（GOT）释放被用作细胞改变的参数。在保存期和复氧期，使用磷酸钠或钾缓冲液时酶释放量最低。使用HEPES和TRIS缓冲液时，在保存期和复氧期观察到LDH和GOT释放率上升。这三种缓冲系统诱导的酶释放与其pKa值相关。保存溶液的较高pH导致细胞中酶泄漏增加。相比之下，低pH的组氨酸/盐酸缓冲液系统在保存期以及复氧期均导致明显的细胞损伤。MOPS是溶液中pH最低的弱酸，在保存期导致最低的酶释放，但在用标准培养基复氧后导致高酶释放。与对照组相比，UW保存后用MOPS孵育培养物导致酶水平较低。总之，在我们的研究中磷酸盐缓冲液（PBS）效果最佳。
