α-酮戊二酸对C2C12细胞谷氨酰胺和氨代谢动力学的影响

Objective To investigate the effect of α-ketoglutarate (α-KG) on cell growth and metabolic kinetics of glutamine and ammonia in C2C12 cell culture. Methods C2C12 cells were cultured in Dulbecco’s Modified Eagle Medium pretreated with adding α-KG (final concentration in media was 0.1mmol/L,1mmol/L,10mmol/L,20mmol/L and 30 mmol/L respectively). Cells cultured in original medium without addition of α-KG served as control. The total cells were counted and the concentration of glutamine and ammonia in the supernatant was determined in cell growth phase. The calculus mathematic model was used to calculate the glutamine and ammonia metabolic kinetics. Results Compared with the control group, the treatment with α-KG at lower concentration (0.1 mM and 1.0 mM) led to the cell growth significantly. In the cells treated with α-KG at higher concentration (20 mM and 30mM), the cell growth was limited. With the cell cultivation the amount of glutamine in the media decreased continuously. In contrast, ammonia concentration increased gradually. Compared with the control group, the specific glutamine consumption was lower in the groups treated with α-KG except for the group at 30mmol/L. However，the specific ammonia production in groups treated at lower α-KG concentration and at higher concentration was respectively lower and higher than that in control group. The general trends among the all the groups, the mean specific ammonia productions for groups were significantly less than that in control groups. There was no great difference of ammonia yield on glutamine between the control group and the addition of α-KG groups. Conclusions: The treatment with α-KG led to profound effects on cell growth. α-KG at reasonable concentration can stimulate cell growth, whereas α-KG at high concentration can impair cell growth. On the other aspect, α-KG treatment led to attenuated specific production of ammonia during the cell culture procedure, which might be attributed to the improved glutamine metabolic efficacy and the stimulant effect on cell growth.