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海洋鱼类奇努克鲑胚胎细胞的生长与代谢:对葡萄糖和谷氨酰胺缺乏的反应。

Growth and metabolism of marine fish Chinook salmon embryo cells: response to lack of glucose and glutamine.

作者信息

Chen Juxing, Sun Xiangming, Zhang Yuanxing

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, 200237, China.

出版信息

Biotechnol Lett. 2005 Mar;27(6):395-401. doi: 10.1007/s10529-005-1774-1.

Abstract

A peculiar phenomenon, differing from the response of mammalian cells, occurred when Chinook salmon embryo (CHSE) cells were passaged in the medium lacking of both glucose and glutamine. To elucidate metabolic mechanism of CHSE cells, the metabolism parameters, key metabolic enzymes, and ATP levels were measured at different glucose and glutamine concentrations. In the glutamine-free culture, hexokinase activity kept constant, and lactate dehydrogenase (LDH) activity decreased. This indicated that lack of glutamine did not expedite glucose consumption but made it shift to lower lactate production and more efficient energy metabolism. The results coincided with the experimental results of unaltered specific glucose consumption rate and decreased yield coefficients of lactate to glucose. In the glucose-free culture, simultaneous increase of glutaminase activity and of specific ammonia production rate suggested an increased flux into the glutaminolysis pathway, and increases of both glutamate dehydrogenase activity and yield coefficient of ammonia to glutamine showed an increased flux into deamination pathway. However, when glucose and glutamine were both lacking, the specific consumption rates of most of amino acids increased markedly, together with decrease of LDH activity, indicating that pyruvate derived from amino acids, away from lactate production, remedied energy deficiency. When both glucose and glutamine were absent, intracellular ATP contents and the energy charge remained virtually unaltered.

摘要

当奇努克鲑鱼胚胎(CHSE)细胞在缺乏葡萄糖和谷氨酰胺的培养基中传代培养时,会出现一种不同于哺乳动物细胞反应的奇特现象。为了阐明CHSE细胞的代谢机制,在不同葡萄糖和谷氨酰胺浓度下测量了代谢参数、关键代谢酶和ATP水平。在无谷氨酰胺培养中,己糖激酶活性保持恒定,而乳酸脱氢酶(LDH)活性降低。这表明缺乏谷氨酰胺不会加快葡萄糖消耗,但会使其转向较低的乳酸生成和更有效的能量代谢。该结果与特定葡萄糖消耗率未改变以及乳酸对葡萄糖的产率系数降低的实验结果一致。在无葡萄糖培养中,谷氨酰胺酶活性和特定氨生成率同时增加表明进入谷氨酰胺分解途径的通量增加,而谷氨酸脱氢酶活性以及氨对谷氨酰胺的产率系数增加表明进入脱氨基途径的通量增加。然而,当葡萄糖和谷氨酰胺都缺乏时,大多数氨基酸的特定消耗率显著增加,同时LDH活性降低,这表明源自氨基酸的丙酮酸不再用于乳酸生成,而是弥补了能量不足。当葡萄糖和谷氨酰胺都不存在时,细胞内ATP含量和能荷实际上保持不变。

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