缺氧状态下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

目的 探讨缺氧状态下肝细胞糖酵解的改变。 方法 配制 3种不同浓度的低氧混合气体 ,用以体外培养大鼠正常肝细胞株BRL ,相应地设为A、B、C组 ,每组设 1、2、4、8、16h 5个缺氧时相点。另以正常肝细胞作为对照。采用生物化学方法 ,检测各时相点肝细胞糖酵解关键酶己糖激酶(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)与正常值比较 ,A、B组BRL细胞的LDH活性在各缺氧时相点均明显增加(P <0 .0 5 ) ,C组细胞的LDH活性从缺氧 2h开始明显增加 (P <0 .0 5 )。 (2 )与正常值比较 ,A组细胞的HK活性在 1、2、4、16h时明显升高 (P <0 .0 5 ) ;B、C两组细胞的HK活性在 1h时明显升高 (P<0 .0 5 )。A组细胞的PFK活性在 1、4h时明显升高 (P <0 .0 5 ) ;B、C两组细胞的PFK活性在 4h时明显升高 (P <0 .0 5 )。 3组细胞的PK活性在各缺氧时相点均明显增高 (P <0 0 5 )。 (3)与正常值比较 ,A、B两组细胞培养液的LA浓度在缺氧 2h时开始增加 (P <0 .0 5 ) ,C组细胞的LA浓度在缺氧 4h时开始增加 (P <0 .0 5 ) ,并均随时间的延长而升高。 结论 缺氧可启动肝细胞糖酵解。

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.