Introduction: Neuroglial functions may be deteriorated in major depressive disorder (MDD). Objective: To evaluate the markers of glial and neuronal cell turnover and to explore their associations with brain metabolites. Methods: In 10 participants with MDD and 10 healthy controls (HC) we investigated neuronal and glial plasma markers (the neuron-specific enolase, NSE; and S100beta, S100B) and brain metabolites (N-acetyl aspartate, NAA; total choline, Cho; and total creatine, Cr). Blood was collected for NSE and S100B. NAA, Cho, and Cr metabolite levels were measured in the anterior cingulate cortex (ACC) with proton magnetic resonance spectroscopy (H-1-MRS) at 3T. Results: NSE and S100B levels were significantly higher in MDD subjects than in HC. The Cr level was significantly higher in MDD subjects than in HC, but the NAA and Cho levels did not differ between groups. NAA/Cr and Cho/Cr ratios were significantly lower in patients with MDD versus HC. S100B was negatively correlated with the Cho levels. Conclusions: These results provide supporting evidence of neuronal and glial distress in MDD. Neuronal viability appears decreased, whereas glial regenerative activity and energy metabolism in the ACC increase in acute major depressive episode. Since low concentrations of S100B have neuroplastic effects, these changes may indicate a possible compensatory mechanism.