Challiss R A, Crabtree B, Newsholme E A
Eur J Biochem. 1987 Feb 16;163(1):205-10. doi: 10.1111/j.1432-1033.1987.tb10756.x.
A method is presented which allows simultaneous estimation of rates of glycogen synthesis and glycogenolysis in an isolated incubated skeletal muscle, thus allowing measurement of the glycogen/glucose-1-phosphate substrate cycling rate. In the absence of hormonal additions, the measured rates of glycogen synthesis and breakdown were similar [respectively, 0.54 +/- 0.12 (8) and 0.74 +/- 0.10 (8) mumol glucosyl equiv. h-1 (g tissue)-1]. Incremental addition of insulin stimulated glycogen synthesis up to threefold and reduced glycogenolysis by about sevenfold; the half-maximally effective concentration of insulin on both processes was about 100 microU/ml (0.7 nM). Incremental addition of adrenaline (in the presence of 1 mU insulin/ml) caused a dramatic increase in the glycogenolytic rate (about 15-fold), but a much less marked inhibition of glycogen synthetic rate. In addition to hormonal manipulation of the muscle preparation in vitro, the effects of cold exposure, the hyperthyroid state, a single exercise bout and exercise-training of animals in vivo on the rates of glycogen synthesis and breakdown in the isolated incubated muscle preparation have been investigated. Significant changes in measured glycogen synthesis, breakdown and glycogen/glucose-1-phosphate cycling have been observed, both under basal conditions and in response to hormonal additions in vitro. The results are discussed with respect to the possible physiological importance of this substrate cycle.
本文介绍了一种方法,该方法可同时估算离体孵育骨骼肌中糖原合成与糖原分解的速率,从而能够测定糖原/葡萄糖-1-磷酸底物循环速率。在未添加激素的情况下,测得的糖原合成与分解速率相似[分别为0.54±0.12(8)和0.74±0.10(8)μmol葡萄糖基当量·h⁻¹·(g组织)⁻¹]。胰岛素的递增添加使糖原合成增加至三倍,并使糖原分解降低约七倍;胰岛素对这两个过程的半数最大有效浓度约为100μU/ml(0.7nM)。肾上腺素的递增添加(在存在1mU胰岛素/ml的情况下)导致糖原分解速率急剧增加(约15倍),但对糖原合成速率的抑制作用则小得多。除了在体外对肌肉制剂进行激素处理外,还研究了冷暴露、甲状腺功能亢进状态、单次运动以及动物体内运动训练对离体孵育肌肉制剂中糖原合成与分解速率的影响。在基础条件下以及体外添加激素后,均观察到测得的糖原合成、分解以及糖原/葡萄糖-1-磷酸循环有显著变化。针对该底物循环可能的生理重要性对结果进行了讨论。