Hauet T, Goujon J M, Tallineau C, Carretier M, Eugene M
Laboratoire de Chirurgie Expérimentale-Institut National de Recherche Agronomique, Surgères and EA 2426, Faculté de Médecine and Centre Hospitalier Universitaire Poitiers, France.
Br J Surg. 1999 Nov;86(11):1401-9. doi: 10.1046/j.1365-2168.1999.01233.x.
Proton nuclear magnetic resonance (NMR) spectroscopy can be used as a non-invasive tool to measure renal damage. In the present investigation, proton NMR spectroscopy of urine was assessed in order to detect cellular damage after different periods of cold ischaemia in two standard preservation solutions.
The isolated perfused pig kidney was used to assess initial renal function after in situ cold flush and cold storage (CS) for 24 or 48 h in two standard preservation solutions: EuroCollins (EC) and University of Wisconsin (UW) solutions. Kidneys flushed with cold heparinized saline and immediately perfused were used as a control group. Kidneys were perfused for 2 h at 37.5 degrees C for functional evaluation. During reperfusion, renal perfusion flow rate was measured. Glomerular filtration rate (GFR), tubular reabsorption of sodium ions, and lactate dehydrogenase (LDH) and N-acetyl-beta-D-glucosaminidase (NAG) excretion were determined. Impairment caused by ischaemia and reperfusion was also determined by histological techniques and proton NMR spectroscopy.
The perfusion flow rate, GFR and tubular reabsorption of sodium were significantly decreased in experimental groups compared with the control group. There was no significant difference between experimental groups after 24 h of CS. The perfusion flow rate was significantly decreased in the EC group after 48 h of cold ischaemia compared with that in the UW group. After 48 h of CS, GFR and tubular reabsorption of sodium were significantly reduced in the EC group compared with those in the UW group. The release of LDH into the effluent and the urinary excretion of NAG were not significantly different after 24 h of CS. After more than 45 and 60 min of reperfusion respectively, LDH and NAG excretion was no different in the 48-h CS groups. The most relevant resonances determined by proton NMR spectroscopy were of citrate, trimethylamine-N-oxide, lactate, acetate and amino acids. Excretion of these markers was significantly more accurate and efficient to assess renal ischaemia-reperfusion injury than that of biochemical markers. A resonance (P) detected particularly in the EC group after 48 h of CS was identified and correlated well with renal dysfunction. After CS for 48 h and 2 h of reperfusion, renal injury was histologically more pronounced in EC groups than in UW groups. However, the difference was not significant after CS for 24 h.
NMR spectroscopy, which is a non-invasive and non-destructive technique, is more accurate and efficient when assessing kidney damage after cold ischaemia and reperfusion when compared to conventional histological and biochemical analysis.
质子核磁共振（NMR）波谱可作为一种非侵入性工具来测量肾损伤。在本研究中，对尿液进行质子NMR波谱分析，以检测在两种标准保存溶液中经历不同时长冷缺血后的细胞损伤。
采用离体灌注猪肾，评估其在两种标准保存溶液（EuroCollins（EC）溶液和威斯康星大学（UW）溶液）中进行原位冷冲洗和冷保存（CS）24或48小时后的初始肾功能。用冷肝素化盐水冲洗并立即灌注的肾脏作为对照组。在37.5℃下对肾脏灌注2小时以进行功能评估。在再灌注过程中，测量肾脏灌注流速。测定肾小球滤过率（GFR）、肾小管对钠离子的重吸收以及乳酸脱氢酶（LDH）和N-乙酰-β-D-氨基葡萄糖苷酶（NAG）的排泄。缺血和再灌注造成的损伤也通过组织学技术和质子NMR波谱进行测定。
与对照组相比，实验组的灌注流速、GFR和肾小管对钠的重吸收显著降低。CS 24小时后，各实验组之间无显著差异。冷缺血48小时后，EC组的灌注流速与UW组相比显著降低。CS 48小时后，EC组的GFR和肾小管对钠的重吸收与UW组相比显著降低。CS 24小时后，流出液中LDH的释放和NAG的尿排泄无显著差异。在再灌注分别超过45和60分钟后，48小时CS组的LDH和NAG排泄无差异。质子NMR波谱测定的最相关共振峰来自柠檬酸盐、氧化三甲胺、乳酸盐、乙酸盐和氨基酸。与生化标志物相比，这些标志物的排泄在评估肾缺血-再灌注损伤方面显著更准确、高效。特别是在CS 48小时后的EC组中检测到的一个共振峰（P）与肾功能障碍密切相关。CS 48小时和再灌注2小时后，EC组的肾损伤在组织学上比UW组更明显。然而，CS 24小时后差异不显著。
NMR波谱作为一种非侵入性和非破坏性技术，与传统的组织学和生化分析相比，在评估冷缺血和再灌注后的肾损伤时更准确、高效。
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