Changani K K, Fuller B J, Bryant D J, Bell J D, Ala-Korpela M, Taylor-Robinson S D, Moore D P, Davidson B R
Department of Surgery, Royal Free Hospital & Medical School, London, UK.
J Hepatol. 1997 Feb;26(2):336-42. doi: 10.1016/s0168-8278(97)80050-5.
We have developed a quick, non-invasive method for measuring the ability of an isolated preserved liver to regenerate high energy phosphate nucleotides without the need for biopsy. Using 31P MRS we have monitored the hepatic energetics of intact cold preserved pig liver using standard clinical harvesting and storage techniques. Following cold storage for 2 h the livers were hypothermically reperfused with oxygenated modified University of Wisconsin preservation fluid. Prior to reperfusion MRS detectable adenosine diphosphate plus adenosine triphosphate was negligible; however, the spectrum showed intense resonances from phosphomonoesters and inorganic phosphate, as a consequence of adenosine triphosphate hydrolysis during cold preservation. Following a 10-min period of hypothermic reperfusion, regeneration of adenosine triphosphate occurred with a concurrent decline in inorganic phosphate and phosphomonoester, both of which are associated with adenosine triphosphate synthesis. The capacity of the liver to regenerate adenosine triphosphate following a 24-h period of cold storage was reduced by approximately 40% (p < 0.01) of the total amount achieved following the shorter cold storage time. Adenosine triphosphate regeneration rates were biphasic and were decreased upon prolonged storage, with the initial rate being reduced from 40.6 x 10(-2).min-1 (standard deviation (sd) 2.70 x 10(-2).min-1) to 14.8 x 10(-2).min-1 (sd; 2.4 x 10(-2).min-1) and the secondary rate from 1.77 x 10(-2).min-1 (sd; 0.18 x 10(-2).min-1) to 0.84 x 10(-2).min-1 (sd; 0.45 x 10(-2).min-1). MR images of the liver during the period of hypothermic reperfusion were also performed providing an assessment for the degree of hepatic vascular perfusion. This non-invasive, 31P MRS assessment of hepatic energetics in a clinically relevant animal model has great potential for the understanding of graft preservation injury.
我们已经开发出一种快速、非侵入性的方法,用于测量离体保存肝脏再生高能磷酸核苷酸的能力,而无需进行活检。我们使用31P磁共振波谱(MRS),采用标准的临床获取和保存技术,监测完整冷保存猪肝的肝脏能量代谢情况。在冷藏2小时后,肝脏用含氧的改良威斯康星大学保存液进行低温再灌注。再灌注前,MRS可检测到的二磷酸腺苷加三磷酸腺苷可忽略不计;然而,由于冷保存期间三磷酸腺苷的水解,光谱显示来自磷酸单酯和无机磷酸盐的强烈共振。在低温再灌注10分钟后,三磷酸腺苷发生再生,同时无机磷酸盐和磷酸单酯减少,这两者都与三磷酸腺苷的合成有关。冷保存24小时后肝脏再生三磷酸腺苷的能力降低,约为较短冷保存时间后所达到总量的40%(p < 0.01)。三磷酸腺苷的再生速率呈双相性,并且随着保存时间的延长而降低,初始速率从40.6×10⁻².min⁻¹(标准差(sd)2.70×10⁻².min⁻¹)降至14.8×10⁻².min⁻¹(sd;2.4×10⁻².min⁻¹),次级速率从1.77×10⁻².min⁻¹(sd;0.18×10⁻².min⁻¹)降至0.84×10⁻².min⁻¹(sd;0.45×10⁻².min⁻¹)。还在低温再灌注期间对肝脏进行了磁共振成像(MR),以评估肝脏血管灌注程度。这种在临床相关动物模型中对肝脏能量代谢进行的非侵入性31P MRS评估,对于理解移植物保存损伤具有巨大潜力。