Simpson D P, Sherrard D J
J Clin Invest. 1969 Jun;48(6):1088-96. doi: 10.1172/JCI106065.
The effect of variations of medium pH and bicarbonate concentration on glutamine oxidation was studied in slices and mitochondria from dog renal cortex. Decreasing pH and bicarbonate concentration increased the rate of oxidation of glutamine-U-(14)C to (14)CO(2) in both slices and mitochondria, an effect comparable to the acute stimulation of glutamine utilization produced by metabolic acidosis. Decreases in the concentration of glutamate and alpha-ketoglutarate, which accompany metabolic acidosis in the intact animal, also occurred in tissue slices when pH and [HCO(3) (-)] were lowered; decrease in alpha-ketoglutarate but not in glutamate content occurred in mitochondria under these conditions. Study of independent variations of medium pH and [HCO(3) (-)] showed that simultaneous changes in both pH and [HCO(3) (-)] produced a greater effect on glutamine metabolism than did change in either of these parameters alone. The rate of glutamine oxidation was also compared in tissue preparations from pairs of litter-mate dogs with chronic metabolic acidosis and alkalosis. No significant difference in the rate of glutamine oxidation was present in mitochondria from the two sets of animals. Slices from animals with chronic metabolic acidosis consistently oxidized glutamine at a more rapid rate than slices from alkalotic dogs both at high and at low concentrations of bicarbonate in the medium. We believe this difference is a result of the same mechanism which leads to the delayed increase in ammonium excretion during induction of metabolic acidosis. The close parallel between the effects demonstrated here and the changes in ammonium production and glutamine utilization in the intact animal with metabolic acidosis suggest that the observed in vitro changes accurately represent the operation of the physiologic mechanism by which acid-base changes regulate ammonium excretion. The similarity between the changes in glutamine oxidation observed in this study and those described previously for citrate suggests that one control mechanism affects the metabolism of both citrate and glutamine. Thus, we believe that the increase in citrate clearance in metabolic alkalosis and the increase in glutamine utilization and ammonium production in metabolic acidosis reflect the operation of the same underlying biochemical mechanism. This mechanism permits changes in pH and [HCO(3) (-)] in the cellular environment to regulate the rate of mitochondrial uptake and oxidation of several physiologically important substrates.
研究了培养基pH值和碳酸氢盐浓度变化对犬肾皮质切片和线粒体中谷氨酰胺氧化的影响。降低pH值和碳酸氢盐浓度会增加切片和线粒体中谷氨酰胺 - U -(14)C氧化为(14)CO₂的速率,这一效应与代谢性酸中毒引起的谷氨酰胺利用急性刺激相当。在完整动物中伴随代谢性酸中毒出现的谷氨酸和α - 酮戊二酸浓度降低,在组织切片中当pH值和[HCO₃⁻]降低时也会发生;在这些条件下,线粒体中α - 酮戊二酸含量降低,但谷氨酸含量未降低。对培养基pH值和[HCO₃⁻]独立变化的研究表明,pH值和[HCO₃⁻]同时变化对谷氨酰胺代谢的影响比单独改变这两个参数中的任何一个都更大。还比较了患有慢性代谢性酸中毒和碱中毒的同窝幼犬组织制剂中谷氨酰胺氧化速率。两组动物的线粒体中谷氨酰胺氧化速率没有显著差异。在培养基中高碳酸氢盐浓度和低碳酸氢盐浓度下,患有慢性代谢性酸中毒动物的切片始终比患有碱中毒动物的切片以更快的速率氧化谷氨酰胺。我们认为这种差异是导致代谢性酸中毒诱导期间铵排泄延迟增加的相同机制的结果。这里所展示的效应与完整动物代谢性酸中毒时铵产生和谷氨酰胺利用变化之间的密切平行关系表明,体外观察到的变化准确地代表了酸碱变化调节铵排泄的生理机制的运作。本研究中观察到的谷氨酰胺氧化变化与先前描述的柠檬酸变化之间的相似性表明,一种控制机制影响柠檬酸和谷氨酰胺的代谢。因此,我们认为代谢性碱中毒时柠檬酸清除率增加以及代谢性酸中毒时谷氨酰胺利用和铵产生增加反映了相同的潜在生化机制的运作。这种机制允许细胞环境中pH值和[HCO₃⁻]的变化来调节线粒体对几种生理重要底物的摄取和氧化速率。