Kletzien R F, Pariza M W, Becker J E, Potter V R, Butcher F R
J Biol Chem. 1976 May 25;251(10):3014-20.
Amino acid transport was studied in primary cultures of parenchymal cells isolated from adult rat liver by a collagenase perfusion technique and maintained as a monolayer in a serum-free culture medium. Amino acid transport was assayed by measuring the uptake of the nonmetabolizable amino acid, alpha-aminoisobutyric acid. Rat liver parenchymal cells transported alpha-aminoisobutyric acid by an energy-dependent Na+-requiring system which displayed Michaelis-Menten kinetics. Addition of insulin to cultured rat liver parenchymal cells resulted in an increased influx of alpha-aminoisobutyric acid which was reflected in a higher initial rate of alpha-aminoisobutyric acid transport as well as an increased accumulation of alpha-aminoisobutyric acid at later time points. Cycloheximide effectively blocked the increase while results with actinomycin D were equivocal. Insulin at concentrations as low as 50 pM was effective in stimulating alpha-aminoisobutyric acid transport while the maximal response was observed at 80 nM.
采用胶原酶灌注技术从成年大鼠肝脏分离实质细胞,并在无血清培养基中维持单层培养,对其氨基酸转运进行了研究。通过测量不可代谢氨基酸α-氨基异丁酸的摄取来测定氨基酸转运。大鼠肝脏实质细胞通过一种依赖能量且需要钠离子的系统转运α-氨基异丁酸,该系统呈现米氏动力学。向培养的大鼠肝脏实质细胞中添加胰岛素会导致α-氨基异丁酸流入增加,这表现为α-氨基异丁酸转运的初始速率更高,以及在后期时间点α-氨基异丁酸的积累增加。放线菌酮有效地阻断了这种增加,而放线菌素D的结果则不明确。低至50 pM的胰岛素浓度即可有效刺激α-氨基异丁酸转运,而在80 nM时观察到最大反应。