GLUCOSE-UPTAKE IN HUMAN AND ANIMAL MUSCLE-CELLS IN CULTURE
GLUCOSE-UPTAKE IN HUMAN AND ANIMAL MUSCLE-CELLS IN CULTURE
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DOI：
10.1139/o90-076
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发表时间：
1990-02-01
影响因子：
2.9
通讯作者：
KLIP, A
中科院分区：
文献类型：
--
作者：
SARABIA, V;RAMLAL, T;KLIP, A
人肌肉细胞在培养物中从肌肉活检中存在的卫星细胞生长，并鉴定融合感受态克隆。在培养的人肌细胞的融合肌管中研究己糖摄取，并与大鼠L 6和小鼠C2 C12肌细胞系的肌管中的己糖摄取进行比较。2-脱氧葡萄糖的摄取是饱和的，并表现出约1.5 mM的所有三种细胞类型的肌管中的表观Km。摄取的Vmax为约6000 pmol/（min. mg蛋白），4000 pmol/（min. mg蛋白），以及500 pmol/（min. mg蛋白）。在人、大鼠和小鼠肌肉细胞培养物中，细胞松弛素B抑制了约90%的己糖摄取。胰岛素刺激2-脱氧葡萄糖摄取在所有三个文化。该激素还刺激3-O-甲基葡萄糖的转运。在人和C2 C12小鼠肌管中对胰岛素的敏感性更高（在3.5 × 10 - 6时观察到半最大刺激）。10-9 M）比在大鼠L 6肌管中（在2.5 × 2.5时观察到的半最大刺激）更强。10-8 M）。然而，胰岛素（10-6 M）刺激己糖摄取在L 6在更大程度上（2.37倍）比人（1.58倍）或小鼠（1.39倍）肌管。它的结论是，在培养中生长的人肌肉细胞显示载体介导的葡萄糖摄取，与其他肌肉细胞的定性相似的特性，胰岛素刺激己糖摄取在人体细胞。这些培养物将有助于研究人类胰岛素抵抗和抗糖尿病药物的作用机制。
Human muscle cells were grown in culture from satellite cells present in muscle biopsies and fusion-competent clones were identified. Hexose uptake was studied in fused myotubes of human muscle cells in culture and compared with hexose uptake in myotubes of the rat L6 and mouse C2C12 muscle cell lines. Uptake of 2-deoxyglucose was saturable and showed an apparent Km of about 1.5 mM in myotubes of all three cell types. The Vmax of uptake was about 6000 pmol/(min .cntdot. mg protein) in human cells, 4000 pmol/(min .cntdot. mg protein) in mouse C2C12 muscle cells, and 500 pmol/(min .cntdot. mg protein) in L6 cells. Hexose uptake was inhibited .apprx.90% by cytochalasin B in human, rat, and mouse muscle cell cultures. Insulin stimulated 2-deoxyglucose uptake in all three cultures. The hormone also stimulated transport of 3-O-methylglucose. The sensitivity to insulin was higher in human and C2C12 mouse myotubes (half-maximal stimulation observed at 3.5 .times. 10-9 M) than in rat L6 myotubes (half-maximal stimulation observed at 2.5 .times. 10-8 M). However, insulin (10-6 M) stimulated hexose uptake to a larger extent (2.37-fold) in L6 than in either human (1.58-fold) or mouse (1.39-fold) myotubes. It is concluded that human muscle cells grown in culture display carrier-mediated glucose uptake, with qualitatively similar characteristics to those of other muscle cells, and that insulin stimulates hexose uptake in human cells. These cultures will be instrumental in the study of human insulin resistance and in investigations on the mechanism of action of antidiabetic drugs.