Active glycolysis and glycogenolysis in early stages of primary cultured hepatocytes. Role of AMP and fructose 2,6-bisphosphate.
This study examines the factors involved in the rapid glycolysis and glycogenolysis that occur during the first stages of hepatocyte culture: a) Shortly after seeding glycolysis, estimated as lactate released to culture medium, increased 10 times in comparison to that reported in vivo. By 8 to 9 h of culture, hepatocytes were nearly glycogen-depleted even in the presence of insulin. b) 6-Phosphofructo-2-kinase remained 100% active during this period. The proportion of the initial active phosphorylase (87%) decreased to 57% by 7 h of culture. c) Fructose 2,6-bisphosphate content was initially similar to that found in liver of fed animals, decreased after seeding and increased thereafter up to four times the initial concentration. In spite of changes in the concentration of this activator, the glycolytic rate remained high and constant. d) ADP and AMP increased sharply after cell plating, reaching values 1.7 and 3.5 times higher. The rise in AMP levels may be involved in the activation of glycolysis and glycogenolysis, because this metabolite is known to act as an allosteric activator of phosphofructokinase and glycogen phosphorylase. This metabolic situation resembles that of cells under hypoxia.
Pilar López
M
M
Centro de Investigación, Hospital La Fe, INSALUD, Valencia, Spain.
Gómez-Lechón
M J
MJ
Castell
J V
JV
D000244:Adenosine Diphosphate / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D005635:Fructosediphosphates / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D006003:Glycogen / Q000378:metabolism*; D006019:Glycolysis; D006598:Hexosediphosphates / Q000378:metabolism*; D007700:Kinetics; D007773:Lactates / Q000096:biosynthesis; D019344:Lactic Acid; D008099:Liver / Q000378:metabolism*; D008297:Male; D010732:Phosphofructokinase-1 / Q000378:metabolism; D006005:Phosphorylases / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains
D005635:Fructosediphosphates; D006598:Hexosediphosphates; D007773:Lactates; D019344:Lactic Acid; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; C027652:fructose 2,6-diphosphate; D000255:Adenosine Triphosphate; D006003:Glycogen; D006005:Phosphorylases; D010732:Phosphofructokinase-1; D005947:Glucose