Selection of endogenous 13C substrates for observation of intracellular metabolism using the dynamic nuclear polarization technique
Selection of endogenous 13C substrates for observation of intracellular metabolism using the dynamic nuclear polarization technique
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DOI：
10.1007/s11604-009-0390-8
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发表时间：
2010-02-10
影响因子：
2.1
通讯作者：
Otsuka, Hideki
中科院分区：
文献类型：
--
作者：
Harada, Masafumi;Kubo, Hitoshi;Otsuka, Hideki
目的.本研究的目的是选择一个合适的底物候选动态核极化（DNP）的研究，并证明其效用评估细胞内代谢。超极化物质包括1-C-13-丙酮酸（Pyr）、1-C-13-葡萄糖（Glc）和1-C-13-乙酸。一个DNP偏振器和600 MHz的垂直小口径扫描仪用于C-13-MR光谱测量。极化1小时后，将溶解的溶液通过毛细管线注入扫描仪中的核磁共振管中。超极化C-13标记的底物的连续光谱在超过120 s的持续时间内获得，并且在此后超过1 h获得热光谱。培养人乳腺癌FM 3A细胞，检测细胞内超极化C-13物质。最大的灵敏度被发现使用Pyr与最长的T1衰减（51.5秒）;和显着的，最小的灵敏度被观察到使用葡萄糖与信号衰减小于2秒。使用其他物质显示出灵敏度的有效增加。使用C-13-Pyr的超极化细胞内研究显示乳酸水平明显升高。DNP技术可用于评价细胞内代谢。然而，Glc不适合与DNP技术一起使用。
Purpose. The aim of this study was to select a suitable substrate candidate for dynamic nuclear polarization (DNP) studies and demonstrate its utility for evaluating intracellular metabolism.Materials and methods. Hyperpolarized substances included 1-C-13-pyruvate (Pyr), 1-C-13-glucose (Glc), and 1-C-13-acetate. A DNP polarizer and a 600-MHz vertical small-bore scanner were used for C-13-MR spectroscopic measurements. After polarization for 1 h, the dissolved solution was injected via a capillary line into the nuclear magnetic resonance tube in the scanner. The sequential spectra of the hyperpolarized C-13-labeled substrates were acquired in durations of more than 120 s, and a thermal spectrum was obtained more than 1 h thereafter. FM3A cancer cells of mammary tumors were cultured for intracellular detection of the hyperpolarized C-13-substances.Results. The greatest sensitivity was found using Pyr with the longest T1 decay (51.5 s); and remarkably, the least sensitivity was observed using Glc with a signal decay of less than 2 s. An effective increase in sensitivity was shown using the other substances. The hyperpolarized intracellular study using C-13-Pyr showed distinct elevation of lactate levels.Conclusion. The DNP technique is useful for evaluating intracellular metabolism. However, Glc is not suitable for use with the DNP technique.