Effects of alpha-ketoglutarate on cell growth, glucose, glutamine, lactate and ammonia metabolism in C2C12 cells
Background: Nutritional supplement with alpha-ketoglutarate (alpha-KG) can lead to improved exercise tolerance. However, mechanisms underlying the phenomenon have not yet been addressed. The purpose of the study was to investigate the effects of alpha-KG on cell growth in C2C12 cell culture and on the energy metabolism and to explore the potential dose-response of these effects to alpha-KG. Methods: C2C12 cells were cultured in DMEM pretreated with adding alpha-KG. The cells and media were harvested every 24 h. From the cell samples the total cells were counted and the substrates and productions metabolism dynamics were evaluated. In addition, a cell clonogenic assay was performed. Results: 1) From the control cells the colony formation efficiency (CFE) was 50. The treatment with alpha-KG at lower concentration (0.1 mM and 1.0 mM) led to a CFE 68 % and 55 %, respectively. At higher concentration (20 mM and 30 mM), the CFE were 10 % and 6 %, respectively. 2) With the cell cultivation the amount of glucose and glutamine in the media decreased continuously. The glucose and glutamine specific consumption rate was significantly lower in the alpha-KG-treated groups than that of the control. 3) The amount of lactate and ammonia increased rapidly with the culture procedure. Compared with the control the lactate and ammonia specific production rate during the exponential growth phase were lower in alpha-KG groups. Conclusions: 1) The treatment with alpha-KG led to profound effects on colony formation, cell growth rate and maximum cell count. alpha-KG at reasonable concentration can stimulate cell growth whereas alpha-KG at high concentration can impair cell growth. 2) alpha-KG treatment resulted in lower specific consumption of glucose and glutamine. 3) alpha-KG treatment led to attenuated specific production of lactate and ammonia during the cell culture procedure. The results of this study might help understand mechanisms underlying the supportive effect of alpha-KG on physical training in human beings.
Subject HeadingsAmmoniakstoffwechsel [GND]