冲洗过程中灌注压力对采用无创荧光测定法获取的肝脏活力的影响。

Effects of perfusion pressure during flushing on the viability of the procured liver using noninvasive fluorometry.

作者信息

Tokunaga Y, Ozaki N, Wakashiro S, Ikai I, Kimoto M, Morimoto T, Shimahara Y, Kamiyama Y, Yamaoka Y, Ozawa K

机构信息

Department of Surgery, Faculty of Medicine, Kyoto University, Japan.

出版信息

Transplantation. 1988 Jun;45(6):1031-5. doi: 10.1097/00007890-198806000-00007.

DOI:10.1097/00007890-198806000-00007

PMID:3289147

Abstract

The influence of perfusion pressure and flow rate on hepatic cellular viability was investigated in the procured liver using noninvasive pyridine nucleotide fluorometry, in relation to other metabolic indices such as nicotinamide adenine dinucleotide (NAD) concentration, adenine nucleotides, tissue ketone bodies, and mitochondrial phosphorylative activity. Rat liver was perfused at a controlled flow rate through the portal vein with a nonrecirculating open-end-design perfusion system driven by a roller pump using cooled oxygenized Euro-Collins' solution. The maximum pressures (mean pressure in parentheses) of experimental groups were 8 (5) mmHg, 15 (9) mmHg, 40 (25) mmHg, 80 (50) mmHg, 120 (73) mmHg, 240 (152) mmHg. The amplitude between oxidation and reduction levels (RxA) in the fluorometric trace, and the gradient or velocity of the trace curve from oxidation to reduction (RxV) were determined by the measurement of fluorescence from NAD phosphate, reduced form (NADPH) using a new fluorometric device. Other metabolic indices were measured after attainment of a fully oxygenated state. RxA decreased in accordance with the increase of pressure. Similar results were obtained in the changes of NAD concentration, total adenine nucleotides, and total tissue ketone bodies, possibly due to the wash-out of these metabolites. RxV decreased in inverse proportion to the increase of pressure. There were close positive correlations between RxV and tissue ketone body ratio (acetoacetate/3-hydroxybutyrate) (r = 0.793, less than 0.01), RxV and respiratory control ratio in isolated mitochondria (r = 0.617, P less than 0.05), RxV and state 3 respiration (r = 0.792, P less than 0.01), and RxV and phosphorylation rate (r = 0.833, P less than 0.01). These results suggest that perfusion pressure and flow rate can easily deteriorate the energy metabolism of the procured liver, and that the gravity-induced hydrostatic pressure presently used in procurement perfusion should be reevaluated on the basis of energy metabolism.

摘要

利用无创吡啶核苷酸荧光测定法，在获取的肝脏中研究灌注压力和流速对肝细胞活力的影响，并与其他代谢指标相关联，这些指标包括烟酰胺腺嘌呤二核苷酸（NAD）浓度、腺嘌呤核苷酸、组织酮体和线粒体磷酸化活性。使用由滚轴泵驱动的非循环开放式灌注系统，以控制的流速通过门静脉灌注大鼠肝脏，灌注液为冷却的充氧Euro - Collins溶液。实验组的最大压力（括号内为平均压力）分别为8（5）mmHg、15（9）mmHg、40（25）mmHg、80（50）mmHg、120（73）mmHg、240（152）mmHg。使用新型荧光测定装置通过测量还原型烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的荧光，来确定荧光测定轨迹中氧化水平与还原水平之间的幅度（RxA）以及轨迹曲线从氧化到还原的斜率或速度（RxV）。在达到完全充氧状态后测量其他代谢指标。RxA随着压力的增加而降低。在NAD浓度、总腺嘌呤核苷酸和总组织酮体的变化中也获得了类似结果，这可能是由于这些代谢物的洗脱所致。RxV与压力的增加成反比降低。RxV与组织酮体比率（乙酰乙酸/3 - 羟基丁酸）之间存在密切的正相关（r = 0.793，P<0.01），RxV与分离线粒体中的呼吸控制比率之间存在密切的正相关（r = 0.617，P<0.05），RxV与状态3呼吸之间存在密切的正相关（r = 0.792，P<0.01），以及RxV与磷酸化速率之间存在密切的正相关（r = 0.833，P<0.01）。这些结果表明，灌注压力和流速很容易使获取的肝脏的能量代谢恶化，并且目前在获取灌注中使用的重力诱导静水压力应基于能量代谢进行重新评估。