Institute of Clinical Physiology, National Research Council (IFC-CNR), 20159 Milano, Italy.
Department of Molecular Medicine, IRCCS Foundation Policlinico San Matteo, 27100 Pavia, Italy.
Int J Mol Sci. 2023 Jan 5;24(2):1029. doi: 10.3390/ijms24021029.
This study addresses a joint nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR) spectroscopy approach to provide a platform for dynamic assessment of kidney viability and metabolism. On porcine kidney models, ROS production, oxidative damage kinetics, and metabolic changes occurring both during the period between organ retrieval and implantation and after kidney graft were examined. The 1H-NMR metabolic profile—valine, alanine, acetate, trimetylamine-N-oxide, glutathione, lactate, and the EPR oxidative stress—resulting from ischemia/reperfusion injury after preservation (8 h) by static cold storage (SCS) and ex vivo machine perfusion (HMP) methods were monitored. The functional recovery after transplantation (14 days) was evaluated by serum creatinine (SCr), oxidative stress (ROS), and damage (thiobarbituric-acid-reactive substances and protein carbonyl enzymatic) assessments. At 8 h of preservation storage, a significantly (p < 0.0001) higher ROS production was measured in the SCS vs. HMP group. Significantly higher concentration data (p < 0.05−0.0001) in HMP vs. SCS for all the monitored metabolites were found as well. The HMP group showed a better function recovery. The comparison of the areas under the SCr curves (AUC) returned a significantly smaller (−12.5 %) AUC in the HMP vs. SCS. EPR-ROS concentration (μmol·g−1) from bioptic kidney tissue samples were significantly lower in HMP vs. SCS. The same result was found for the NMR monitored metabolites: lactate: −59.76%, alanine: −43.17%; valine: −58.56%; and TMAO: −77.96%. No changes were observed in either group under light microscopy. In conclusion, a better and more rapid normalization of oxidative stress and functional recovery after transplantation were observed by HMP utilization.
本研究采用联合磁共振(NMR)和电子顺磁共振(EPR)光谱学方法,为动态评估肾脏活力和代谢提供了一个平台。在猪肾模型中,研究了器官获取和植入期间以及移植后肾移植物发生的 ROS 产生、氧化损伤动力学和代谢变化。监测了 1H-NMR 代谢谱(缬氨酸、丙氨酸、醋酸盐、三甲胺-N-氧化物、谷胱甘肽、乳酸)和 EPR 氧化应激(缺血再灌注损伤后由静态冷藏(SCS)和离体机器灌注(HMP)方法保存 8 小时引起的)。通过血清肌酐(SCr)、氧化应激(ROS)和损伤(硫代巴比妥酸反应性物质和蛋白质羰基酶)评估来评估移植后的功能恢复。在保存 8 小时时,SCS 组的 ROS 产生显著(p < 0.0001)高于 HMP 组。还发现,与 SCS 相比,所有监测代谢物的 HMP 组浓度数据(p < 0.05-0.0001)均显著升高。HMP 组显示出更好的功能恢复。比较 SCr 曲线下面积(AUC)的结果表明,HMP 组的 AUC 明显较小(-12.5%)。HMP 组活检肾组织样本中的 EPR-ROS 浓度(μmol·g−1)显著低于 SCS 组。NMR 监测的代谢物也得到了相同的结果:乳酸:-59.76%;丙氨酸:-43.17%;缬氨酸:-58.56%;TMAO:-77.96%。在任何一组中,在光镜下均未观察到变化。总之,通过使用 HMP,观察到移植后氧化应激和功能恢复的更好和更快的正常化。