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

METHODSMixed 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).

CONCLUSIONhypoxia might initiate glycolysis.