Comparison of protection by fructose against paracetamol injury with protection by glucose and fructose-1,6-diphosphate

Abstract

We have compared the protective effect of fructose in normal Ringer solution during the onset and progression of cell injury induced by paracetamol in rat liver slices with the protective effect of glucose and fructose-1,6-diphosphate. Liver slices obtained from phenobarbitone-induced and non-induced rats were used in a model in vitro system. Slices were exposed to 10 mM paracetamol for 120 min and then incubated without paracetamol in the presence or absence of protective agents for a further 240 min. Cell injury was quantified by measuring leakage of lactate dehydrogenase (LDH) and potassium (K +). Adenosinetriphosphate (ATP) levels were measured using the luciferin-luciferase bioluminescence assay. Addition of higher concentrations of glucose (10–50 mM) to Ringer solution were not found to result in protection at the end of incubation in paracetamol-treated slices obtained from phenobarbitone-induced rats. Neither did sucrose nor mannitol protect. However, exclusion of glucose from Ringer solution resulted in cell injury in paracetamol-treated slices obtained from non-induced rats. Methionine, a known antidote for paracetamol poisoning, failed to protect in this instance but fructose did protect. This suggests that the presence of a glycolytic substrate plays a crucial role in cell protection. Further evidence for this is the finding that iodoacetate, an inhibitor of glycolysis, not only increases cell injury in paracetamol-treated slices but also reverses fructose protection. Fructose-1, 6-diphosphate was found to protect against the onset and progression of cell injury in paracetamol-treated slices obtained from phenobarbitone induced rats. This protective agent is found to maintain high ATP levels and cell viability in paracetamol-treated slices at a time when paracetamol-treated slices show a profound loss of ATP levels and a significant increase in cell injury as measured by leakage of LDH and K +.