Simultaneous measurement of NAD metabolome in aged mice tissue using liquid chromatography tandem-mass spectrometry
Simultaneous measurement of NAD metabolome in aged mice tissue using liquid chromatography tandem-mass spectrometry
复制标题
DOI：
10.1002/bmc.4205
复制
发表时间：
2018-06-01
影响因子：
1.8
通讯作者：
Nakagawa, Takashi
中科院分区：
文献类型：
--
作者：
Yaku, Keisuke;Okabe, Keisuke;Nakagawa, Takashi
烟酰胺腺嘌呤二核苷酸(NAD)是介导氧化还原反应和基因表达等多种生物学过程的主要辅因子。最近，NAD的代谢受到了相当大的关注，因为给予NAD前体对动物衰老相关的代谢紊乱具有良好的效果。虽然已有大量研究报道NAD在多种动物组织中的水平随着年龄的增长而下降，但NAD在衰老器官中的代谢途径和动力学尚不完全清楚。为了确定NAD在衰老时的代谢，我们建立了基于LC/MS/MS系统的靶向代谢组学。我们的方法简单，适用于粗制的生物样品，包括培养细胞和动物组织。与传统的酶循环分析不同，我们的方法可以通过执行单个样品制备来确定NAD和NADH(NAD的简化形式)。此外，我们使用生物样本验证了我们的方法，并研究了NAD代谢组在老化过程中的变化。与以前的报告一致，肝脏和骨骼肌中的NAD水平随着年龄的增长而下降。此外，我们发现随着年龄的增长，肾脏中烟酰胺单核苷酸和烟酰胺核苷的含量显著增加。我们建立的基于LC/MS/MS的NAD代谢组学广泛应用于生物医学研究，其结果将为衰老研究，特别是NAD代谢研究提供创新思路。
Nicotinamide adenine dinucleotide (NAD) is a major co-factor that mediates multiple biological processes including redox reaction and gene expression. Recently, NAD metabolism has received considerable attention because administration of NAD precursors exhibited beneficial effects against aging-related metabolic disorders in animals. Although numerous studies have reported that NAD levels decline with aging in multiple animal tissues, the pathway and kinetics of NAD metabolism in aged organs are not completely understood. To determine the NAD metabolism upon aging, we developed targeted metabolomics based on an LC/MS/MS system. Our method is simple and applicable to crude biological samples, including culture cells and animal tissues. Unlike a conventional enzymatic cycling assay, our approach can determine NAD and NADH (reduced form of NAD) by performing a single sample preparation. Further, we validated our method using biological samples and investigated the alteration of the NAD metabolome during aging. Consistent with previous reports, the NAD levels in the liver and skeletal muscle decreased with aging. Further, we detected a significant increase in nicotinamide mononucleotide and nicotinamide riboside in the kidney upon aging. The LC/MS/MS-based NAD metabolomics that we have developed is extensively applicable to biomedical studies, and the results will present innovative ideas for the aging studies, especially for that of NAD metabolism.