Laboratory of Analytical and Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, and Global COE Program, University of Shizuoka, Suruga-ku, Shizuoka, Japan.
J Proteome Res. 2010 Aug 6;9(8):3912-22. doi: 10.1021/pr100121k.
The number of diabetic patients has recently been increasing worldwide. Thus, the discovery of potential diabetic biomarker(s), leading to the early detection and/or prevention of diabetes mellitus, is strongly required. The diagnosis of the prediabetic state in humans is a very difficult issue because of the lifestyle differences in each person and ethical consideration. Upon the basis of these considerations, animal experiments using ddY strain mice (ddY-H), which undergo naturally occurring diabetes along with age, were carried out in this study. Biomarker discovery based upon a metabonome study is now quite common, the same as that in the proteome analysis. Reversed-phase liquid chromatography-mass spectrometry (LC-MS) has mainly been used for the extensive analysis of low-molecular mass compounds including metabolites. The metabolites in the plasma of diabetic mice (ddY-H) and normal mice (ddY-L) were exhaustively separated and detected by ultraperformance liquid chromatography along with electrospray ionization time-of-flight mass spectrometry (UPLC-ESI-TOF-MS) using T3-C18 and HS-F5 columns. The biomarker candidates related to diabetes mellitus were extracted from the metabolite profiling of ddY-H and ddY-L at 5, 9 13, and 20 weeks old using a multivariate statistical analysis such as orthogonal partial least-squares-discriminant analysis (OPLS-DA). Various metabolites and unknown compounds were detected as biomarker candidates related to diabetic mellitus. Furthermore, the concentration of several metabolites on Lysine biosynthesis and Lysine degradation pathways were remarkably changed between the 9-week old ddY-H and ddY-L mice. Because a couple of biomarker candidates related to the prediabetic state were identified using the present approach, the metabolite profiling study could be helpful for understanding the abnormal state of various diseases.
近年来，全球糖尿病患者的数量一直在增加。因此，强烈需要发现潜在的糖尿病生物标志物，以实现糖尿病的早期检测和/或预防。由于每个人的生活方式不同和伦理考虑，人类糖尿病前期状态的诊断是一个非常困难的问题。基于这些考虑，本研究使用自然发生糖尿病的 ddY 品系小鼠（ddY-H）进行了动物实验。基于代谢组学研究的生物标志物发现现在非常普遍，与蛋白质组分析相同。反相液相色谱-质谱联用（LC-MS）主要用于广泛分析低分子量化合物，包括代谢物。通过超高效液相色谱-电喷雾电离飞行时间质谱联用（UPLC-ESI-TOF-MS）使用 T3-C18 和 HS-F5 柱，对糖尿病小鼠（ddY-H）和正常小鼠（ddY-L）血浆中的代谢物进行了详尽的分离和检测。使用正交偏最小二乘判别分析（OPLS-DA）等多变量统计分析方法，从 5、9、13 和 20 周龄的 ddY-H 和 ddY-L 的代谢物图谱中提取与糖尿病相关的生物标志物候选物。检测到各种代谢物和未知化合物作为与糖尿病相关的生物标志物候选物。此外，在 9 周龄的 ddY-H 和 ddY-L 小鼠之间，赖氨酸生物合成和赖氨酸降解途径上的几种代谢物的浓度明显发生变化。由于使用本方法鉴定了与糖尿病前期状态相关的一些生物标志物候选物，因此代谢物图谱研究可能有助于了解各种疾病的异常状态。
