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D-葡萄糖对大鼠肝细胞中D-果糖代谢的影响:一项13C核磁共振研究。

Effects of D-glucose upon D-fructose metabolism in rat hepatocytes: A 13C NMR study.

作者信息

Malaisse W J, Ladrière L, Verbruggen I, Willem R

出版信息

Mol Cell Biochem. 2002 Dec;241(1-2):103-6. doi: 10.1023/a:1020818810935.

Abstract

Isolated hepatocytes from fed rats were exposed for 120 min to D-glucose (10 mM) and either D-[1-13C]fructose, D-[2-13C]fructose or D-[6-13C]fructose (also 10 mM) in the presence of D2O. The identification and quantification of 13C-enriched D-fructose and its metabolites (D-glucose, L-lactate, L-alanine) in the incubation medium and the measurement of their deuterated isotopomers indicated, by comparison with a prior study conducted in the absence of exogenous D-glucose, that the major effects of the aldohexose were to increase the recovery of 13C-enriched D-fructose, decrease the production of 13C-enriched D-glucose, restrict the deuteration of the 13C-enriched isotopomers of D-glucose to those generated by cells exposed to D-[2-13C]fructose, and to accentuate the lesser deuteration of the C, (as compared to C5) of 13C-enriched D-glucose derived from D-[2-13C]fructose. The ratio between C2-deuterated and C2-hydrogenated L-lactate, as well as the relative amounts of the CH3-, CH2D-, CHD, and CD3- isotopomers of 13C-enriched L-lactate were not significantly different, however, in the absence or presence of exogenous D-glucose. These findings indicate that exogenous D-glucose suppressed the deuteration of the C1 of D-[I-13C]glucose generated by hepatocytes exposed to D-[1-13C]fructose or D-[6-13C]fructose, as otherwise attributable, in part at least, to gluconeogenesis from fructose-derived [3-13C]pyruvate, and apparently favoured the phosphorylation of D-fructose by hexokinase isoenzymes, probably through stimulation of D-fructose phosphorylation by glucokinase.

摘要

将喂食后的大鼠分离出的肝细胞,在重水存在的情况下,分别与D-葡萄糖(10 mM)以及D-[1-¹³C]果糖、D-[2-¹³C]果糖或D-[6-¹³C]果糖(均为10 mM)一起孵育120分钟。通过与之前在无外源性D-葡萄糖条件下进行的研究相比较,对孵育培养基中¹³C富集的D-果糖及其代谢产物(D-葡萄糖、L-乳酸、L-丙氨酸)进行鉴定和定量,并测量它们的氘代同位素异构体,结果表明,己醛糖的主要作用是提高¹³C富集的D-果糖的回收率,降低¹³C富集的D-葡萄糖的生成量,将¹³C富集的D-葡萄糖同位素异构体的氘代限制为细胞暴露于D-[2-¹³C]果糖时产生的那些异构体,并加剧源自D-[2-¹³C]果糖的¹³C富集的D-葡萄糖的C1(与C5相比)的较低氘代。然而,在有无外源性D-葡萄糖的情况下,C2氘代与C2氢化的L-乳酸之间的比率,以及¹³C富集的L-乳酸的CH₃-、CH₂D-、CHD-和CD₃-同位素异构体的相对量没有显著差异。这些发现表明,外源性D-葡萄糖抑制了肝细胞暴露于D-[1-¹³C]果糖或D-[6-¹³C]果糖时产生的D-[1-¹³C]葡萄糖C1的氘代,这在其他情况下至少部分可归因于果糖衍生的[3-¹³C]丙酮酸的糖异生作用,并且显然有利于己糖激酶同工酶对D-果糖的磷酸化,可能是通过刺激葡萄糖激酶对D-果糖的磷酸化来实现的。

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