Piccioni Fabiana, Borioni Anna, Delfini Maurizio, Del Giudice Maria Rosaria, Mustazza Carlo, Rodomonte Andrea, Risuleo Gianfranco
Department of Chemistry, University of Rome La Sapienza, P.le A. Moro 5, 00185 Rome, Italy.
Anal Biochem. 2007 Aug 1;367(1):111-21. doi: 10.1016/j.ab.2007.04.013. Epub 2007 Apr 12.
Proton nuclear magnetic resonance (NMR) spectroscopy was used to identify and quantify the metabolites present in cultured mouse fibroblast cells 3T6 in their native state and after treatment with PD166866, an inhibitor of the fibroblast growth factor receptor. Cell extracts were prepared according to the Bligh-Dyer protocol which prevents artifacts deriving from the chemical demolition of macromolecules. Also the growth medium was subjected to the same extraction procedure. The NMR approach made possible the identification and quantification of about 40 different metabolites at nanomoles/mg of protein level: the biological relevance of the variation of some metabolite levels is discussed. Our experimental procedure offers a prospective method for the evaluation of variations of the metabolic profile deriving from different biochemical treatments of these cells.
质子核磁共振（NMR）光谱法用于鉴定和定量天然状态下以及用成纤维细胞生长因子受体抑制剂PD166866处理后的培养小鼠成纤维细胞3T6中存在的代谢物。根据Bligh-Dyer方案制备细胞提取物，该方案可防止因大分子化学分解而产生的假象。生长培养基也采用相同的提取程序。NMR方法能够在纳摩尔/毫克蛋白质水平上鉴定和定量约40种不同的代谢物：讨论了一些代谢物水平变化的生物学相关性。我们的实验程序为评估这些细胞不同生化处理后代谢谱的变化提供了一种前瞻性方法。
