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[缺氧时大鼠肝细胞糖酵解变化的实验研究]

[An experimental study of the changes of rat hepatocytic glycolysis during hypoxia].

作者信息

Ma Zhengwei, Wang Shiliang, Wang Fengjun, Wang Pei

机构信息

Institute of Burn Research, Southwestern Hospital, Third Military Medical University, Chongqing 400038, P.R. China.

出版信息

Zhonghua Shao Shang Za Zhi. 2002 Aug;18(4):238-41.

Abstract

OBJECTIVE

To investigate the effects of hypoxia on the glycolysis in cultured rat hepatocytes.

METHODS

Mixed gas with different concentrations of O(2), CO(2) and N(2) was prepared for the in vitro culture of normal rat hepatocytes. The cell strains were set to be A, B, C groups, which were observed at 1, 2, 4, 8 and 16 hours after hypoxia with normal hepatocytes as the control. Biochemical methods were employed to determine the activities of the key enzymes during hepatocytic glycolysis such as hexokinase (HK), phosphofructokinase (PFK), pyruvate kinase (PK), lactate dehydrogenase (LDH) and the change of the content of lactic acid (LA) in the culture fluid.

RESULTS

(1) The LDH activity of the rat hepatocytes increased significantly at all the time points of hypoxia in A and B groups when compared with that in control group (P < 0.05), while the activity increased obviously in C group since 2 hours after hypoxia (P < 0.05). (2) The HK activity of the cells in A group increased significantly at 1, 2, 4 and 16 hours after hypoxia and that in B and C groups increased obviously at 1 hour when compared with control group (P < 0.05). While the cellular PFK activity in A group increased markedly at 1 and 4 hours after hypoxia and that in B and C groups increased evidently at 4 hours after hypoxia (P < 0.05). The cellular PK activity in all the three groups increased at all the hypoxic time points (P < 0.05). (3) The cellular LA content in A and B groups began to increase since 2 hours and that in C group did so since 4 hours after hypoxia and increased along with the time lapse (P < 0.05).

CONCLUSION

hypoxia might initiate glycolysis.

摘要

目的

研究缺氧对培养的大鼠肝细胞糖酵解的影响。

方法

制备含不同浓度O₂、CO₂和N₂的混合气体用于正常大鼠肝细胞的体外培养。将细胞株设为A、B、C组,以正常肝细胞为对照,在缺氧后1、2、4、8和16小时进行观察。采用生化方法测定肝细胞糖酵解过程中关键酶如己糖激酶(HK)、磷酸果糖激酶(PFK)、丙酮酸激酶(PK)、乳酸脱氢酶(LDH)的活性以及培养液中乳酸(LA)含量的变化。

结果

(1)与对照组相比,A组和B组大鼠肝细胞在缺氧各时间点的LDH活性均显著升高(P<0.05),而C组自缺氧2小时后活性明显升高(P<0.05)。(2)A组细胞在缺氧后1、2、4和16小时的HK活性显著升高,B组和C组在缺氧1小时时活性明显升高(P<0.05)。A组细胞在缺氧后1和4小时的PFK活性显著升高,B组和C组在缺氧4小时时活性明显升高(P<0.05)。三组细胞的PK活性在所有缺氧时间点均升高(P<0.05)。(3)A组和B组细胞在缺氧2小时后LA含量开始增加,C组在缺氧4小时后开始增加,并随时间推移而升高(P<0.05)。

结论

缺氧可能启动糖酵解。

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