α-酮戊二酸对C2C12细胞谷氨酰胺和氨代谢动力学的影响.docVIP

  α-酮戊二酸对 C2C12 细胞谷氨酰胺和氨代 谢动力学的影响# 杨兵全1,3，孙子林1,3，李玲1,3，金晖1,3，张林2，李凤飞1,3** 5 10 15 （1. 东南大学附属中大医院内分泌科，南京 210009； 2. 东南大学临床医学院， 南京 210009； 3. 东南大学糖尿病研究所，南京 210009） 摘要：目的 研究α-酮戊二酸对体外培养的 C2C12 细胞生长及谷氨酰胺和氨代谢动力学的 影响。方法 用含不同浓度α-酮戊二酸的 DMEM 培养液（0.1mmol/L，1mmol/L，10mmol/L， 20mmol/L 和 30mmol/L）干预体外培养的 C2C12 细胞，用不含α-酮戊二酸的 DMEM 培养 液作为对照，观察 C2C12 细胞的增殖，培养液中谷氨酰胺和氨代谢动力学变化。结果 在培 养液中添加适度浓度的α-酮戊二酸能促进 C2C12 细胞的增殖。然而，高浓度的α-酮戊二酸 能抑制 C2C12 细胞的增殖。在细胞增殖期，α-酮戊二酸添加组细胞特异性谷氨酰胺消耗量 较对照组减少。与对照组相比，在培养液中添加α-酮戊二酸能显著减少氨的产生，同时对 氨谷氨酰胺的产率无显著影响。 结论 在细胞培养液中添加适度浓度的α-酮戊二酸能刺激 C2C12 细胞的生长，增加了细胞对谷氨酰胺的利用效率，并减少氨的产生。 关键词：α-酮戊二酸；C2C12 细胞；细胞培养；氨代谢；谷氨酰胺代谢 中图分类号：R34 20 Effect of α-ketoglutarate on the metabolism kinetics of glutamine and ammonia in C2C12 cells Yang Bingquan1,3, Sun Zilin1,3, Li Ling1,3, Jin Hui1,3, Zhang Lin2, Li Fengfei1,3 (1. Endocrinology Section, Southeast University Zhongda Hospital, Nanjing 210009; 25 30 35 40 45 2. Medicine School, Southeast University, Nanjing 210009; 3. Diabetes Institute, Southeast University, Nanjing 210009) Abstract: 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 de