This study investigated the neuroprotective effect of glucose metabolites against oxygen-glucose deprivation in primary cultured neurons. Pure neurons were isolated and cultured from cerebral neocortices of embryonic mice at 15 days of gestation. Mature cells grown for 7, 14, or 21 days after dissection were exposed to oxygen- glucose deprivation (OGD) and supplemented with various concentrations of glucose metabolites. It was confirmed that the level of neuronal injury was proportional to the duration of exposure to hypoxia, and inversely proportional to oxygen concentration. In particular, older cells (21 days after culture) were exceedingly susceptible to hypoxic/ anoxic injury. However, when supplemented with moderately high concentration of glucose (11 mM) or pyruvate (12 mM) the injury from OGD was significantly attenuated. Our results demonstrate the possibility of pyruvate being a favorable alternative to glucose during OGD. Accordingly, a proteomic analysis via 2-D gel electrophoresis was done with protein extracts from pyruvate-treated cells after OGD injury. 1500 spots were identified from image analysis, revealing a total of 47 proteins standing out with significantly different abundance after pyruvate administration, compared to cells injured with no supplement. This differential expression was most prominent in those with size around 66 kDa and pI values around 5.6. Especially, MMP-1 was identified via MALDI- TOF mass spectrometry to be one of the significantly upregulated proteins after pyruvate treatment.

Keywords: proteomics, neuronal cell death, oxygen-glucose deprivation, hing glucose, pyruvate