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灌注大鼠肝脏中戊糖循环的通量率。一种根据[1-14C]葡萄糖产生的14CO2计算底物通量率的方法。

Rates of flux through the pentose cycle in perfused rat liver. A procedure for the calculation of rates of substrate flux from 14CO2 production from [1-14C]glucose.

作者信息

Kuehn A, Scholz R

出版信息

Eur J Biochem. 1982 Jun;124(3):611-7. doi: 10.1111/j.1432-1033.1982.tb06638.x.

Abstract

A method for the determination of substrate flux through the pentose cycle was developed employing [1-14C]glucose in experiments with perfused rat livers. The method consists first of a kinetic analysis which differentiates between the production of 14CO2 from [1-14C]glucose via the pentose cycle and via the citrate cycle and, second of a calculation of the specific radioactivity of the hexose monophosphate pool from measured rates of glycolysis and the specific radioactivity of lactate released into the perfusate. The method was validated by experiments comparing the results of tracer infusions with [1-14C]glucose, [6-14C]glucose and [3-14C]pyruvate. In livers from fed rats perfused with 10 mM glucose, the rate of substrate flux through the pentose cycle was around 0.2 mumol X min-1 X g-1; it was about 20% of the substrate flux via glycolysis. The kinetic data were inconsistent with the existence of an L-type pentose cycle in liver.

摘要

利用[1-14C]葡萄糖,在灌注大鼠肝脏的实验中开发了一种测定通过戊糖循环的底物通量的方法。该方法首先包括动力学分析,该分析区分了[1-14C]葡萄糖通过戊糖循环和柠檬酸循环产生14CO2的情况,其次包括根据测量的糖酵解速率和释放到灌注液中的乳酸的比放射性来计算己糖单磷酸池的比放射性。通过比较用[1-14C]葡萄糖、[6-14C]葡萄糖和[3-14C]丙酮酸进行示踪剂输注的结果的实验,验证了该方法。在用10 mM葡萄糖灌注的喂食大鼠的肝脏中,通过戊糖循环的底物通量速率约为0.2 μmol·min-1·g-1;它约为通过糖酵解的底物通量的20%。动力学数据与肝脏中存在L型戊糖循环不一致。

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