人体尸体肾脏低温机器灌注期间灌注液的代谢组学分析

Metabolomic analysis of perfusate during hypothermic machine perfusion of human cadaveric kidneys.

作者信息

Guy Alison J, Nath Jay, Cobbold Mark, Ludwig Christian, Tennant Daniel A, Inston Nicholas G, Ready Andrew R

机构信息

1 Department of Renal Surgery, University Hospitals Birmingham, Birmingham, United Kingdom. 2 Department of Immunity and Infection, University of Birmingham, Birmingham, United Kingdom. 3 Biomolecular NMR Spectroscopy, University of Birmingham, Birmingham, United Kingdom. 4 School of Cancer Sciences, University of Birmingham, Birmingham, United Kingdom.

出版信息

Transplantation. 2015 Apr;99(4):754-9. doi: 10.1097/TP.0000000000000398.

DOI:10.1097/TP.0000000000000398

PMID:25222017

Abstract

BACKGROUND

The metabolic processes occurring within the preserved kidney during hypothermic machine perfusion (HMP) are not well characterized. The aim of this study was to use nuclear magnetic resonance (NMR) spectroscopy to examine the metabolomic profile of HMP perfusate from human cadaveric kidneys awaiting transplantation and to identify possible discriminators between the profiles of kidneys with delayed graft function (DGF) and immediate graft function (IGF).

METHODS

Perfusates from HMP kidneys were sampled at 45 min and 4 hr of preservation with the LifePort Kidney Transporter 1.0 (Organ Recovery Systems, Chicago, IL) using KPS-1. Prepared samples underwent 1-D Proton-NMR spectroscopy, and resultant spectra were analyzed. Clinical parameters were collected prospectively.

RESULTS

Perfusate of 26 transplanted cadaveric kidneys was analyzed; 19(73%) with IGF and 7(27%) with DGF. Glucose concentrations were significantly lower in DGF kidneys compared to those with IGF at both 45 min (7.772 vs. 9.459 mM, P = 0.006) and 4 hr (8.202 vs. 10.235 mM, P = 0.003). Concentrations of inosine and leucine were significantly different between DGF and IGF kidneys at 45 min (0.002 vs. 0.013 mM, P = 0.009 and 0.011 vs. 0.006 mM, P = 0.036), and gluconate levels were also significantly different between DGF and IGF kidneys at 4 hr (49.099 vs. 59.513 mM, P = 0.009).

CONCLUSION

Significant metabolic activity may be occurring in kidneys during HMP. The NMR spectroscopy of the perfusate can identify differences in the metabolomic profiles of DGF and IGF kidneys that might have a predictive role in viability assessment. Modification of harmful metabolic processes may improve outcomes for HMP kidneys.

摘要

背景

低温机器灌注（HMP）期间保存肾内发生的代谢过程尚未得到充分表征。本研究的目的是使用核磁共振（NMR）光谱法检查等待移植的人类尸体肾HMP灌注液的代谢组学特征，并确定移植肾功能延迟（DGF）和移植肾功能即刻恢复（IGF）的肾之间可能的鉴别指标。

方法

使用KPS-1，通过LifePort肾脏转运器1.0（器官回收系统公司，伊利诺伊州芝加哥）在保存45分钟和4小时时从HMP肾中采集灌注液样本。制备的样本进行一维质子NMR光谱分析，并对所得光谱进行分析。前瞻性收集临床参数。

结果

分析了26个移植尸体肾的灌注液；19个（73%）移植肾功能即刻恢复，7个（27%）移植肾功能延迟。在45分钟（7.772对9.459 mM，P = 0.006）和4小时（8.202对10.235 mM，P = 0.003）时，移植肾功能延迟的肾中葡萄糖浓度显著低于移植肾功能即刻恢复的肾。在45分钟时，次黄嘌呤和亮氨酸浓度在移植肾功能延迟和移植肾功能即刻恢复的肾之间有显著差异（0.002对(0.013) mM，P = 0.009和0.011对0.006 mM，P = 0.036），4小时时葡萄糖酸盐水平在移植肾功能延迟和移植肾功能即刻恢复的肾之间也有显著差异（49.099对59.513 mM，P = 0.009）。