1H-NMR Studies of Metabolic Effects of Endo- and Exogeneous Compounds on Astroglia Cells Cultivated In Vitro

A 1H-NMR method is described for the study of metabolic reactions in-vitro of neonatal cerebellar rat astroglia cells cultivated on micro carrier beads. In the NMR tube, the formation of metabolic effects on astroglia cells caused by addition of nutrients (pyruvate and fumarate) or neurotoxins can be monitored. Metabolic changes of both endogenous metabolites and toxins, 1,2-epoxybutane and ethyl acrylate are investigated. 1,2,4-Benzenetricarboxylate (BTC) served as an extracellular reference. Dy(ATP)-1 was used to depress signals from extracellular compounds. 6-Diazo-5-oxo-Lnorleucin was added to cause accumulation of conjugated intracellular metabolites to an extent that permitted detection by 1H-NMR spectroscopy. 1,2-Epoxybutane is shown to be detoxified due to a transferase reaction involving conjugation of reduced glutathione. The sulphur is coupled to carbon-1 or carbon 2 of the epoxide. Addition of ethyl acrylate leads in a similar way to a C1-GSH derivative. The metabolic status of the astroglia cells were monitored during different exposure conditions.