Nissim I, Yudkoff M, Nissim I, States B
Division of Biochemical Development and Molecular Diseases, Children's Hospital, Philadelphia, Pennsylvania.
J Cell Physiol. 1989 Mar;138(3):536-40. doi: 10.1002/jcp.1041380313.
The present investigation evaluates the effect of AICA-Riboside on the synthesis of purine nucleotides and the growth of normal rat kidney cells in culture. Experiments in the presence and absence of various concentrations of AICA-Riboside were conducted with Dulbecco's Modified Eagle's Medium supplemented with either 1 mM [15N]aspartate or [14N]aspartate. Addition of 50 microM AICA-Riboside to the incubation medium significantly stimulated intracellular adenine nucleotide concentrations following incubation for 48 hours. This stimulation was associated with augmented cell growth and DNA concentration. In contrast, with concentrations above 100 microM of AICA-Riboside in the incubation medium, there was a remarkable inhibition of cell growth and a significant depletion of intracellular pools of adenine nucleotides and DNA. Experiments with [15N]aspartate showed that the initial rate (0-24 hours) of [6-15NH2]adenine nucleotide formation from 1 mM [15N]aspartate was 38.8 +/- 9.6, 67.9 +/- 12.5, and 20.1 +/- 3.8 pmol h-1/10(6) cells in the presence of 0 (control), 50 microM and 500 microM AICA-Riboside, respectively. These observations indicate that the main effect of AICA-Riboside is on the formation of AMP from aspartate and IMP via the sequential action of adenylosuccinate synthetase and adenylosuccinate lyase. The current studies suggest that AICA-Riboside could be used as a factor mediating renal cell mitosis in culture. AICA-Riboside has a biphasic effect on the growth of renal epithelial cells in culture and on their intracellular purine nucleotides and DNA concentration.
本研究评估了5'-氨基咪唑-4-甲酰胺核苷（AICA-核苷）对嘌呤核苷酸合成以及培养的正常大鼠肾细胞生长的影响。在含有不同浓度AICA-核苷的情况下，使用补充了1 mM [15N]天冬氨酸或[14N]天冬氨酸的杜尔贝科改良伊格尔培养基进行实验。在孵育培养基中添加50 microM AICA-核苷，孵育48小时后显著刺激了细胞内腺嘌呤核苷酸浓度。这种刺激与细胞生长增加和DNA浓度升高有关。相比之下，在孵育培养基中AICA-核苷浓度高于100 microM时，细胞生长受到显著抑制，细胞内腺嘌呤核苷酸池和DNA显著减少。用[15N]天冬氨酸进行的实验表明，在0（对照）、50 microM和500 microM AICA-核苷存在的情况下，由1 mM [15N]天冬氨酸形成[6-15NH2]腺嘌呤核苷酸的初始速率（0 - 24小时）分别为38.8 +/- 9.6、67.9 +/- 12.5和20.1 +/- 3.8 pmol h-1/10(6)细胞。这些观察结果表明，AICA-核苷的主要作用是通过腺苷酸琥珀酸合成酶和腺苷酸琥珀酸裂解酶的顺序作用，从天冬氨酸和次黄嘌呤核苷酸形成腺苷酸。当前研究表明，AICA-核苷可作为培养中介导肾细胞有丝分裂的一个因子。AICA-核苷对培养的肾上皮细胞生长及其细胞内嘌呤核苷酸和DNA浓度具有双相作用。
