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分析大鼠原代肝细胞和人肝癌细胞系 Huh7 的氨代谢。

Analysis of the ammonia metabolism of rat primary hepatocytes and a human hepatocyte cell line Huh 7.

机构信息

International Center for Biotechnology, Osaka University, 2-1, Yamada-oka, Suita, Osaka 565, Japan,

出版信息

Cytotechnology. 2000 Jan;32(1):9-15. doi: 10.1023/A:1008165027319.

Abstract

Ammonia metabolism of ratprimary hepatocytes and a human hepatocyte cell line,Huh 7, at different concentrations of glutamine,glucose and ammonia was examined. During theincubation of the primary hepatocyte cells, glutamineand ammonia concentrations decreased, that of ureaincreased, and that of glucose remained the same. Inthe case of Huh 7 cells, glucose was consumed rapidly,the concentration of ammonia increased and that of urearemained the same. The major energy sources amongmedium components were glutamine for the primary cellsand glucose for Huh 7 cells, although the primaryhepatocytes may utilize intracellular glycogen asenergy source. As the glutamine concentration in theincubation medium increased, the specific rates of notonly glutamine consumption, but also ammonia productionby the primary cells and Huh 7 cells increased. Besides, specific urea production rate by the primarycells increased then. Increase of glucoseconcentration had no effect on glutamine and ammoniametabolism by both cells, although it increased glucoseconsumption by Huh 7 cells. The incubation of theprimary cells with higher ammonia concentrationincreased all specific rates of glutamine consumption,ammonia consumption and urea production. An increasein the ammonia concentration to 5 mM changed theammonia metabolism from production to consumption andincreased the specific glucose consumption rate. Consequently, increases in the glutamine and ammoniaconcentrations were revealed to have negative andpositive effects, respectively, on decreasing ammoniaconcentration by both of rat primary hepatocytes andHuh 7 cells.

摘要

研究了不同浓度谷氨酰胺、葡萄糖和氨对大鼠原代肝细胞和人肝癌细胞系 Huh7 氨代谢的影响。在原代肝细胞的孵育过程中,谷氨酰胺和氨浓度降低,尿素浓度增加,葡萄糖浓度保持不变。在 Huh7 细胞中,葡萄糖被迅速消耗,氨浓度增加,尿素浓度保持不变。虽然原代肝细胞可能利用细胞内糖原作为能量来源,但在培养基成分中,谷氨酰胺是原代细胞的主要能量来源,葡萄糖是 Huh7 细胞的主要能量来源。随着孵育培养基中谷氨酰胺浓度的增加,原代细胞和 Huh7 细胞的谷氨酰胺消耗和氨生成的比速率均增加。此外,原代细胞的尿素生成比速率也随之增加。虽然 Huh7 细胞的葡萄糖消耗增加,但增加葡萄糖浓度对两种细胞的谷氨酰胺和氨代谢没有影响。原代细胞在较高氨浓度下孵育会增加所有谷氨酰胺消耗、氨消耗和尿素生成的比速率。将氨浓度增加到 5mM 会使氨代谢由生成变为消耗,并增加特异性葡萄糖消耗率。因此,增加谷氨酰胺和氨浓度分别对大鼠原代肝细胞和 Huh7 细胞降低氨浓度有负向和正向作用。

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