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雏鸡肠道刷状缘细胞中的谷氨酸、谷氨酰胺、天冬氨酸、天冬酰胺、葡萄糖和酮体代谢

Glutamate, glutamine, aspartate, asparagine, glucose and ketone-body metabolism in chick intestinal brush-border cells.

作者信息

Porteous J W

出版信息

Biochem J. 1980 Jun 15;188(3):619-32. doi: 10.1042/bj1880619.

DOI:10.1042/bj1880619
PMID:7470024
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1161942/
Abstract
  1. Suspensions of isolated chick jejunal columnar absorptive (brush-border) cells respired on endogenous substrates at a rate 40% higher than that shown by rat brush-border cells. 2. Added d-glucose (5 or 10mm), l-glutamine (2.5mm) and l-glutamate (2.5mm) were the only individual substrates which stimulated respiration by chick cells; l-aspartate (2.5 or 6.7mm), glutamate (6.7mm), glutamine (6.7mm), l-alanine (1 or 10mm), pyruvate (1 or 2mm), l-lactate (5 or 10mm), butyrate (10mm) and oleate (1mm) did not stimulate chick cell respiration; l-asparagine (6.7mm) inhibited slightly; glucose (5mm) stimulated more than did 10mm-glucose. 3. Acetoacetate (10mm) and d-3-hydroxybutyrate (10mm) were rapidly consumed but, in contrast to rat brush-border cells, did not stimulate respiration. 4. Glucose (10mm) was consumed more slowly than 5mm-glucose; the dominant product of glucose metabolism during vigorous respiration was lactate; the proportion of glucose converted to lactate was greater with 10mm- than with 5mm-glucose. 5. Glutamate and aspartate consumption rates decreased, and alanine and glutamine consumption rates increased when their initial concentrations were raised from 2.5 to 6.7 or 10mm. 6. The metabolic fate of glucose was little affected by concomitant metabolism of any one of aspartate, glutamate or glutamine except for an increased production of alanine; the glucose-stimulated respiration rate was unaffected by concomitant metabolism of these individual amino acids. 7. Chick cells produced very little alanine from aspartate and, in contrast to rat cells, likewise produced very little alanine from glutamate or glutamine; in chick cells alanine appeared to be predominantly a product of transmination of pyruvate derived from glucose metabolism. 8. In chick cells, glutamate and glutamine were formed from aspartate (2.5 or 6.7mm); aspartate and glutamine were formed from glutamate (2.5mm) but only aspartate from 6.7mm-glutamate; glutamate was the dominant product formed from glutamine (6.7mm) but aspartate only was formed from 2.5mm-glutamine. 9. Chick brush-border cells can thus both catabolize and synthesize glutamine; glutamine synthesis is always diminished by concomitant metabolism of glucose, presumably by allosteric inhibition of glutamine synthetase by alanine. 10. Proline was formed from glutamine (2.5mm) but not from glutamine (2.5mm)+glucose (5mm) and not from 2.5mm-glutamate; ornithine was formed from glutamine (2.5mm)+glucose (5.0mm) but not from glutamine alone; serine was formed from glutamine (2.5mm)+glucose (5mm) and from these two substrates plus aspartate (2.5mm). 11. Total intracellular adenine nucleotides (22mumol/g dry wt.) remained unchanged during incubation of chick cells with glucose. 12. Intracellular glutathione (0.7-0.8mm) was depleted by 40% during incubation of respiring chick cells without added substrates for 75min at 37 degrees C; partial restoration of the lost glutathione was achieved by incubating cells with l-glutamate+l-cysteine+glycine.
摘要
  1. 分离出的鸡空肠柱状吸收(刷状缘)细胞悬液以内源性底物进行呼吸的速率比大鼠刷状缘细胞高40%。2. 添加的d-葡萄糖(5或10mmol/L)、l-谷氨酰胺(2.5mmol/L)和l-谷氨酸(2.5mmol/L)是仅有的能刺激鸡细胞呼吸的单一底物;l-天冬氨酸(2.5或6.7mmol/L)、谷氨酸(6.7mmol/L)、谷氨酰胺(6.7mmol/L)、l-丙氨酸(1或10mmol/L)、丙酮酸(1或2mmol/L)、l-乳酸(5或10mmol/L)、丁酸盐(10mmol/L)和油酸盐(1mmol/L)均未刺激鸡细胞呼吸;l-天冬酰胺(6.7mmol/L)有轻微抑制作用;5mmol/L葡萄糖比10mmol/L葡萄糖对呼吸的刺激作用更强。3. 乙酰乙酸(10mmol/L)和d-3-羟基丁酸盐(10mmol/L)被快速消耗,但与大鼠刷状缘细胞不同的是,它们并未刺激呼吸。4. 10mmol/L葡萄糖的消耗速度比5mmol/L葡萄糖慢;在剧烈呼吸过程中,葡萄糖代谢的主要产物是乳酸;10mmol/L葡萄糖转化为乳酸的比例比5mmol/L葡萄糖更高。5. 当初始浓度从2.5mmol/L提高到6.7或10mmol/L时,谷氨酸和天冬氨酸的消耗速率降低,而丙氨酸和谷氨酰胺的消耗速率增加。6. 天冬氨酸、谷氨酸或谷氨酰胺中的任何一种伴随代谢对葡萄糖的代谢命运影响不大,只是丙氨酸的生成增加;这些单个氨基酸的伴随代谢对葡萄糖刺激的呼吸速率没有影响。7. 鸡细胞从天冬氨酸生成的丙氨酸很少,与大鼠细胞不同的是,从谷氨酸或谷氨酰胺生成的丙氨酸也很少;在鸡细胞中,丙氨酸似乎主要是葡萄糖代谢产生的丙酮酸转氨作用的产物。8. 在鸡细胞中,天冬氨酸(2.5或6.7mmol/L)可生成谷氨酸和谷氨酰胺;谷氨酸(2.5mmol/L)可生成天冬氨酸和谷氨酰胺,但6.7mmol/L谷氨酸只能生成天冬氨酸;谷氨酰胺(6.7mmol/L)生成的主要产物是谷氨酸,但2.5mmol/L谷氨酰胺只能生成天冬氨酸。9. 因此,鸡刷状缘细胞既能分解代谢谷氨酰胺,也能合成谷氨酰胺;葡萄糖的伴随代谢总是会使谷氨酰胺的合成减少,推测是丙氨酸对谷氨酰胺合成酶的变构抑制作用所致。10. 脯氨酸由谷氨酰胺(2.5mmol/L)生成,但不由谷氨酰胺(2.5mmol/L)+葡萄糖(5mmol/L)生成,也不由2.5mmol/L谷氨酸生成;鸟氨酸由谷氨酰胺(2.5mmol/L)+葡萄糖(5.0mmol/L)生成,但不由单独的谷氨酰胺生成;丝氨酸由谷氨酰胺(2.5mmol/L)+葡萄糖(5mmol/L)以及这两种底物加天冬氨酸(2.5mmol/L)生成。11. 在鸡细胞与葡萄糖一起孵育的过程中,细胞内总腺嘌呤核苷酸(22μmol/g干重)保持不变。12. 在37℃下,未添加底物的呼吸鸡细胞孵育75分钟后,细胞内谷胱甘肽(0.7 - 0.8mmol/L)减少了40%;用l-谷氨酸+l-半胱氨酸+甘氨酸孵育细胞可部分恢复损失的谷胱甘肽。

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