Glucose loading during primary culture has opposite effects on the viability of hepatocytes exposed to potassium cyanide and to iodoacetic acid

Whether or not to apply nutritional pretreatment and how to do so are contr oversial issues with respect to the liver about to undergo aggressive inter vention. We studied the effects of glucose loading on the viability of hepa tocytes that were subsequently exposed to the inhibitors of carbohydrate me tabolism, potassium cyanide (KCN) and iodoacetic acid (IAA). After rat hepa tocytes were cultured for 24 hours in Leibovitz's L-15 medium containing 0, 10, 20, and 30 mmol/L glucose, the medium was replaced with modified Hanks -HEPES buffer with or without 2.5 mmol/L KCN or 0.5 mmol/L IAA. Lactate deh ydrogenase (LDH) activity, lactate concentration, and pH of the supernatant were measured after 0, 2, 4, and 6 hours of exposure to KCN and after 0, 2 0, 40, and 60 minutes of exposure to IAA. Glycogen and adenosine triphospha te (ATP) contents in the hepatocytes were measured simultaneously. Hepatocy tes cultured with various concentrations of glucose for 24 hours stored Lev els of glycogen in proportion to the glucose concentration in the culture m edium without any significant difference in viability. The hepatocytes cult ured with higher glucose concentrations maintained a higher ATP content and released less LDH and more lactate, and the pH decreased in the supernatan t during exposure to KCN. Conversely, hepatocytes cultured with lower gluco se concentrations maintained a higher ATP content and released less LDH dur ing exposure to IAA. In conclusion, prior glucose loading appears to be ben eficial for hepatocytes if oxidative phosphorylation is to be inhibited, wh ereas withholding glucose appears to be beneficial if glycolysis is to be i nhibited. Copyright (C) 2001 by W.B. Saunders Company.