Klöppel K, Gerlach J, Neuhaus P
Chirurgische Klinik und Poliklinik, Universitätsklinikum Rudolf Virchow, Freie Universität, Berlin.
Langenbecks Arch Chir. 1994;379(4):210-7. doi: 10.1007/BF00186360.
The cation and anion content in liver preservation solutions have been investigated in order to justify the use of "intracellular" or "extracellular" electrolyte compositions. Various concentrations of sodium and potassium with chloride or lactobionate as anions and with added calcium and/or magnesium were made up as preservation solutions and incubated with in vitro adherent cultures of pig hepatocytes. In vitro hypoxia and hypothermia (4 degrees C, PO2 < 0.1 mmHg) for 24 h, with reoxygenation for 3 h, in standard culture medium was used as a model for preservation. Measurements of cell viability and detachment rate by light microscopy and of LDH and GOT liberation were used as parameters of cell damage. Cell swelling was estimated in suspension cultures of isolated hepatocytes. When chloride was used as the anion, significant cell toxicity from potassium concentrations over 75 mM was found within 6 h of preservation. Enzyme liberation decreased with increasing content of sodium cations in the preservation solution. Calcium ions had a protective effect at a concentration of 0.8 mM. Addition of magnesium to an "intracellular" ion composition minimized the toxic effect of potassium cations. Using lactobionate as an impermeant anion, there was no difference between the sodium and the potassium salt and the choice of cation had no effect on enzyme leakage or cell volume. An "extracellular" solution with high sodium chloride content and 0.8 mM calcium resulted in better preservation than was obtained with lactobionate solutions. With chloride as the anion, a significant increase in cell swelling was found when potassium replaced sodium in the solutions. Cell swelling decreased with increasing concentration of sodium cations.(ABSTRACT TRUNCATED AT 250 WORDS)
为了证明使用“细胞内”或“细胞外”电解质组成的合理性,对肝脏保存液中的阳离子和阴离子含量进行了研究。制备了各种浓度的以氯离子或乳糖酸盐为阴离子,并添加了钙和/或镁的钠和钾溶液作为保存液,并与猪肝细胞的体外贴壁培养物一起孵育。以在标准培养基中进行24小时的体外缺氧和低温(4℃,PO2<0.1mmHg),再复氧3小时作为保存模型。通过光学显微镜测量细胞活力和脱离率以及乳酸脱氢酶(LDH)和谷草转氨酶(GOT)的释放作为细胞损伤的参数。在分离的肝细胞悬浮培养物中评估细胞肿胀。当使用氯离子作为阴离子时,在保存6小时内发现钾浓度超过75mM会产生明显的细胞毒性。随着保存液中钠阳离子含量的增加,酶释放减少。钙离子在浓度为0.8mM时具有保护作用。向“细胞内”离子组成中添加镁可将钾阳离子的毒性作用降至最低。使用乳糖酸盐作为非渗透性阴离子时,钠盐和钾盐之间没有差异,阳离子的选择对酶泄漏或细胞体积没有影响。具有高氯化钠含量和0.8mM钙的“细胞外”溶液比乳糖酸盐溶液具有更好的保存效果。当溶液中的钾取代钠时,以氯离子为阴离子会导致细胞肿胀显著增加。细胞肿胀随着钠阳离子浓度的增加而减少。(摘要截短至250字)